Proteolytically stable u1-agatoxin-ta1b variant polypeptides for pest control

Abstract

The application concerns insecticidal proteins and peptides called TVP, which are non-natural peptide variations based upon the U1-agatoxin-Talb toxin derived from the Hobo spider, leading to proteolytically stable variants of the U1-agatoxin-Ta1b. The application also provides polynucleotides and genes encoding these TVPs and their expression in plants; also methods of producing these nucleotides and peptides, processes, production techniques, compositions and formulations of these TVPs, combinations thereof, combinations comprising TVP genes and / or peptides and, organisms expressing these TVPs. These products are useful for the control of pests.

Description

277702-566574 PROTEOLYTICALLY STABLE U1-AGATOXIN-TA1B VARIANT POLYPEPTIDES FOR PEST CONTROL CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of, and priority to, United StatesProvisional Application Serial No.63 / 637,796, filed on April 23, 2024, the contents of which are herein incorporated by reference in their entirety. SEQUENCE LISTING

[0002] This application incorporates by reference in its entirety the Sequence ListingXML entitled “277702-566574.xml” (124,203 bytes), which was created on April 18, 2025, and filed electronically herewith. TECHNICAL FIELD

[0003] New insecticidal proteins, nucleotides, peptides, their expression in plants,methods of producing the peptides, new processes, production techniques, new peptides, new formulations, and combinations of new and known organisms that produce greater yields than would be expected of related peptides for the control of insects are described and claimed. BACKGROUND

[0004] Numerous insects are vectors for disease. Mosquitoes in the genus Anophelesare the principle vectors of Zika virus, Chikungunya virus, and malaria, a disease caused by protozoa in the genus Trypanosoma. Aedes aegypti is the main vector of the viruses that cause Yellow fever and Dengue. Other viruses, the causal agents of various types of encephalitis, are also carried by Aedes spp. mosquitoes. Wuchereria bancrofti and Brugia malayi, parasitic roundworms that cause filariasis, are usually spread by mosquitoes in the genera Culex, Mansonia, and Anopheles.

[0005] Horse flies and deer flies may transmit the bacterial pathogens of tularemia(Pasteurella tularensis) and anthrax (Bacillus anthracis), as well as a parasitic roundworm (Loa loa) that causes loiasis in tropical Africa.

[0006] Eye gnats in the genus Hippelates can carry the spirochaete pathogen thatcauses yaws (Treponema pertenue), and may also spread conjunctivitis (pinkeye). Tsetse flies in the genus Glossina transmit the protozoan pathogens that cause African sleeping sickness (Trypanosoma gambiense and T. rhodesiense). Sand flies in the genus Phlebotomus are vectors of a bacterium (Bartonella bacilliformis) that causes Carrion's disease (Oroyo fever)277702-566574 in South America. In parts of Asia and North Africa, they spread a viral agent that causes sand fly fever (pappataci fever) as well as protozoan pathogens (Leishmania spp.) that cause Leishmaniasis. SUMMARY

[0007] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula I: (all of the variable letters Xn and Bn and Zn are each independently amino acids in single letter format) X1-X2-X3-X4-X5-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A- E-C-F-R-N-D-V-Y-B2-A-C-Z1-Z2-Z3-Z4-Z5-Z6 (SEQ ID NO: 98); wherein X1 is E, or absent, X2is P or absent, X3is D or absent, X4is E, or absent, X5is I or absent, B1is R or Q, B2is T or A; Z1 can be H or absent; Z2 is E or absent; Z3 is A or absent; Z4 is Q or absent; Z5 is K or absent; and Z6is G or absent, and wherein the compound of Formula I does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10) along the entire sequence of the TVP compound.

[0008] In some embodiments, present disclosure provides for U1-agatoxin-Ta1bvariant polypeptide (TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that277702-566574 is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula II: (all of the variable letters Xnand Bnare each independently amino acids in single letter format) X1-X2-X3-X4-X5-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A- E-C-F-R-N-D-V-Y-B2-A-C-H-E-A-Q-K-G (SEQ ID NO: 99); wherein X1 is E, or absent, X2 is P or absent, X3is D or absent, X4is E, or absent, X5is I or absent, B1is R or Q, and B2is T or A; and wherein the compound of Formula II does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10).

[0009] In some embodiments, present disclosure provides for U1-agatoxin-Ta1bvariant polypeptide (TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula III, or an agriculturally acceptable salt thereof: (all of the variable letters Zn and Bn are each independently amino acids in single letter format) E-P-D- E-I-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A-E-C-F-R-N-D- V-Y-B2-A-C-Z1-Z2-Z3-Z4-Z5-Z6 (SEQ ID NO: 100); wherein B1 is R or Q, and B2 is T or A; Z1 is H or absent; Z2 is E or absent; Z3 is A or absent; Z4 is Q or absent; Z5 is K or absent; and Z6is G or absent; and wherein the compound of Formula III does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID277702-566574 NO: 3); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10).

[0010] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure comprises, consists essentially of, or consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15);277702-566574 Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31);277702-566574 Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47);277702-566574 Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50).

[0011] In various embodiments, the TVPs described herein have been shown to havea knockdown of 50% of the population concentration (KD50) of lower than 182 pmol / g against houseflies at 4 hours.

[0012] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13);277702-566574 Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29);277702-566574 Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45);277702-566574 Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions. The TVPs described herein have been shown to have a knockdown of 50% of the population concentration (KD50) of lower than 182 pmol / g against houseflies at 4 hours.

[0013] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 1: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 5.

[0014] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 2: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 6.

[0015] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 3: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 7.

[0016] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 4: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 8.

[0017] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 9);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 9.

[0018] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11);wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 11.

[0019] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 8: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 12.

[0020] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 9: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 13.

[0021] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 14.

[0022] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 15.

[0023] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 12: PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO:277702-566574 16); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 16.

[0024] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 13: DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 17.

[0025] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 14 EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 18.

[0026] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 15: ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 19.

[0027] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 16: CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 20.

[0028] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 17: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 21.

[0029] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 18: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 22.

[0030] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 19: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 23.

[0031] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 20: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 24.

[0032] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 21: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 25.

[0033] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26);wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 26.

[0034] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 27.

[0035] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 28.

[0036] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 29.

[0037] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 30.

[0038] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 31.

[0039] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 32.

[0040] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 33.

[0041] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 34.

[0042] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 35.

[0043] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 36.

[0044] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 37.

[0045] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 34; ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 38.

[0046] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 35; PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 39.

[0047] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 36 DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40);277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 40.

[0048] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 41.

[0049] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE; SEQ ID NO: 42; wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 42.

[0050] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH; SEQ ID NO: 43;277702-566574 wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 43.

[0051] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH; SEQ ID NO: 44; wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 44.

[0052] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH; SEQ ID NO: 45; wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 45.

[0053] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH; SEQ ID NO: 46; wherein the277702-566574 amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 46.

[0054] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC; SEQ ID NO: 47; wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 47.

[0055] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC; SEQ ID NO: 48; wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 48.

[0056] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC; SEQ ID NO: 49; wherein the277702-566574 amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 49.

[0057] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC; SEQ ID NO: 50; wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 50.

[0058] In addition, the present disclosure describes a composition comprising a TVP,and one or more excipients; wherein the TVP comprises an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13);277702-566574 Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29);277702-566574 Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45);277702-566574 Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; and wherein the one or more excipients is selected from the group consisting of: trehalose; maltodextrin; maltose; potassium phosphate dibasic (K2HPO4); potassium phosphate monobasic (KH2PO4); lignosulfonate; gypsum; sorbitol; sodium benzoate; potassium sorbate; EDTA; benzisothiazolinone (BIT); and fermentation solids.

[0059] In addition, the present disclosure describes a composition comprising a TVP,and one or more excipients; wherein the TVP comprises an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12);277702-566574 Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28);277702-566574 Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44);277702-566574 Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; wherein the composition comprises an amount of TVP that is 8.5% wt / wt of the total weight of the composition; and wherein the plurality of excipients consists of the following: an amount of trehalose that is 25% wt / wt; an amount of BIT that is 0.05% wt / wt; an amount of maltodextrin that is 36.3% wt / wt; an amount of potassium phosphate dibasic anhydrous (K2HPO4) that is 2.6% wt / wt; an amount of potassium phosphate monobasic (KH2PO4) that is 0.4% wt / wt; and an amount of fermentation solids that is 26.85% wt / wt, of the total weight of the composition.

[0060] In addition, the present disclosure describes a polynucleotide encoding a TVP,the TVP comprises an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);277702-566574Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25);277702-566574 Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41);277702-566574 Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; or a complementary nucleotide sequence thereof.

[0061] In addition, the present disclosure describes a method of producing a TVP, themethod comprising: (a) preparing a vector comprising a first expression cassette comprising a polynucleotide operable to encode a TVP, or complementary nucleotide sequence thereof, said TVP comprises an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7);277702-566574 Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24);277702-566574 Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40);277702-566574 Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; (b) introducing the vector into a host cell; and (c) growing the host cell in a growth medium under conditions operable to enable expression of the TVP and secretion into the growth medium.

[0062] In addition, the present disclosure describes a method of using thecompositions described above to control insects comprising, providing the compositions to the locus of an insect.

[0063] In addition, the present disclosure describes a method of protecting a plantfrom insects comprising, providing a plant which expresses a TVP, or polynucleotide encoding the same.

[0064] In addition, the present disclosure describes a method of combating,controlling, or inhibiting a pest comprising, applying a pesticidally effective amount of the compositions described above to the locus of the pest, or to a plant or animal susceptible to an attack by the pest.277702-566574

[0065] In addition, the present disclosure describes a vector comprising apolynucleotide operable to encode a TVP having an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19);277702-566574 Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35);277702-566574 Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions.

[0066] In addition, the present disclosure describes a yeast strain comprising: a firstexpression cassette comprising a polynucleotide operable to encode a TVP, said TVP277702-566574 comprising an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20);277702-566574 Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36);277702-566574 Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; or complementary nucleotide sequence thereof. BRIEF DESCRIPTION OF THE DRAWINGS277702-566574

[0067] FIG. 1 depicts the results of a housefly injection assay where activity of Wild-type Ta1b, and the Ta1b Variant Polypeptides (TVPs) with amino acid substitutions at positions R9Q and R9N, were injected intrathoracically into houseflies, and the dose of TVP required for 50% knockdown (KD50) was then determined 24-hours post-injection.

[0068] FIG. 2A shows the changes in compound melting temperatures determined byα-helical content with circular dichroism. The phospholipid headgroup-mimicking compound phytic acid (PA) induces moderate thermal stabilization of SEQ ID NO: 4 and gives an enhanced, higher potency effect for Compound 4.

[0069] FIG. 2B depicts the results of a lipid panel screen assay using P-6001 PIPstrip (Echelon Biosciences, Utah), which contains 15 membrane lipid components including LPA, LPC, PI, PI(3)P, PI(4)P, PI(5)P, PE, PC, S1P, PI(3,4)P2, PI(3, 5)P2, PI(4, 5)P2, P(3, 4, 5)P3, PA and PS.

[0070] FIG. 2C shows the quantification of band density on the lipid panel forPhosphatidic acid (PA).

[0071] FIG. 2D shows the quantification of band density on the lipid panel forPhosphatidylserine (PS).

[0072] FIG. 3A depicts two-electrode voltage clamp (TEVC) recording of applicationof SEQ ID NO: 4 to recombinant Drosophila melanogaster nAChR α4 / β1 receptors expressed in Xenopus oocytes.

[0073] FIG. 3B depicts two-electrode voltage clamp (TEVC) recording of applicationof Compound 1 (SEQ ID NO: 5) to recombinant Drosophila melanogaster nAChR α4 / β1 receptors expressed in Xenopus oocytes.

[0074] FIG. 3C depicts two-electrode voltage clamp (TEVC) recording of applicationof Compound 2 (SEQ ID NO: 6) to recombinant Drosophila melanogaster nAChR α4 / β1 receptors expressed in Xenopus oocytes.

[0075] FIG. 3D depicts two-electrode voltage clamp (TEVC) recording of applicationof Compound 3 (SEQ ID NO: 7) to recombinant Drosophila melanogaster nAChR α4 / β1 receptors expressed in Xenopus oocytes.

[0076] FIG. 3E depicts two-electrode voltage clamp (TEVC) recording of applicationof Compound 4 (SEQ ID NO: 8) to recombinant Drosophila melanogaster nAChR α4 / β1 receptors expressed in Xenopus oocytes.

[0077] FIG. 3F depicts a bar graph showing average potentiation ratios of the SEQID NO: 4 and Compounds 1-4. * = p = 0.0067277702-566574 DETAILED DESCRIPTION

[0078] DEFINITIONS

[0079] “5’-end” and “3’-end” refers to the directionality, i.e., the end-to-endorientation of a nucleotide polymer (e.g., DNA). The 5’-end of a polynucleotide is the end of the polynucleotide that has the fifth carbon.

[0080] “5’- and 3’-homology arms” or “5’ and 3’ arms” or “left and right arms” refersto the polynucleotide sequences in a vector and / or targeting vector that homologously recombine with the target genome sequence and / or endogenous gene of interest in the host organism in order to achieve successful genetic modification of the host organism’s chromosomal locus.

[0081] “Administering” or “administration” or “administer” means to dispense and / orotherwise provide, and refers to any method of application or route of administration. For example, administering can refer to, e.g., application of an insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), or an agriculturally acceptable salt thereof; application of TVP- insecticidal protein or an agriculturally acceptable salt thereof; or a composition comprising a TVP and / or a TVP-insecticidal protein and / or or an agriculturally acceptable salt thereof, and one or more excipients, e.g., a sprayable composition, a foam; a burning formulation; a fabric treatment; a surface-treatment; a dispersant; a microencapsulation, and the like. By “co- application” or “co-administer” it is meant that two or more components are applied or administered at the same time; or a one or more components are applied or administered just prior to, or just after the application the other one or more components. For example, in some embodiments, a first TVP and a second TVP, wherein the first and second TVP can be the same or different, can be applied or administered separately, sequentially, simultaneously, concurrently or chronologically staggered.

[0082] “ADN1 promoter” refers to the DNA segment comprised of the promotersequence derived from the Schizosaccharomyces pombe adhesion defective protein 1 gene.

[0083] “Affect” refers to how a something influences another thing, e.g., how apeptide, polypeptide, protein, drug, or chemical influences an insect, e.g., a pest.

[0084] “Agent” refers to one or more chemical substances, molecules, nucleotides,polynucleotides, peptides, polypeptides, proteins, toxins, toxicants, poisons, insecticides, pesticides, organic compounds, inorganic compounds, prokaryote organisms, or eukaryote organisms (and the agents produced from said prokaryote or eukaryote organisms).277702-566574

[0085] “Agriculturally-acceptable carrier” covers all adjuvants, inert components,dispersants, surfactants, tackifiers, binders, etc. that are ordinarily used in pesticide Formulation technology; these are well known to those skilled in pesticide Formulation.

[0086] “Agriculturally acceptable salt” is synonymous with pharmaceuticallyacceptable salt, and as used herein refers to a compound that is modified by making acid or base salts thereof. In some embodiments, agriculturally acceptable salts, hydrates, solvates, crystal forms and individual isomers, enantiomers, tautomers, diastereomers and prodrugs of the TVPs described herein can be utilized. Non-limiting examples of agriculturally acceptable salts include: acid addition salts, formed with inorganic acids; acid addition salts formed with organic acids; or salts formed when an acidic proton present in the parent compound is replaced by a metal ion, e.g., an alkali metal ion, aluminum ion; or coordinates with an organic base such as ethanolamine, and the like. Other non-limiting examples of agriculturally acceptable salts include: hydrochloride; sodium; sulfate; acetate; phosphate or diphosphate; chloride; potassium; maleate; calcium; citrate; mesylate; nitrate; tartrate; aluminum; gluconate; any organic or inorganic addition salt; alkali metal salts (sodium salts, potassium salts, etc.); alkaline earth metal salts (calcium salts, magnesium salts, etc.); salts derived from inorganic or organic bases (e.g., sodium, potassium, lithium, ammonium, calcium, magnesium, ferrous, zinc, copper, manganous, aluminum, ferric, manganic salts, isopropylamine, diethylamine, ethanolamine, piperidine, tromethamine, choline, and the like).

[0087] “Agroinfection” means a plant transformation method where DNA isintroduced into a plant cell by using Agrobacteria tumefaciens or Agrobacteria rhizogenes.

[0088] “Alignment” refers to a method of comparing two or more sequences (e.g.,nucleotide, polynucleotide, amino acid, peptide, polypeptide, or protein sequences) for the purpose of determining their relationship to each other. Alignments are typically performed by computer programs that apply various algorithms, however, it is also possible to perform an alignment by hand. Alignment programs typically iterate through potential alignments of sequences and score the alignments using substitution tables, employing a variety of strategies to reach a potential optimal alignment score. Commonly-used alignment algorithms include, but are not limited to, CLUSTALW (see Thompson J. D., Higgins D. G., Gibson T. J., CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice, Nucleic Acids Research 22: 4673-4680, 1994); CLUSTALV (see Larkin M. A., et al., CLUSTALW2, ClustalW and ClustalX version 2, Bioinformatics 23(21): 2947-2948, 2007);277702-566574 Mafft; Kalign; ProbCons; and T-Coffee (see Notredame et al., T-Coffee: A novel method for multiple sequence alignments, Journal of Molecular Biology 302: 205-217, 2000). Exemplary programs that implement one or more of the foregoing algorithms include, but are not limited to, MegAlign from DNAStar (DNAStar, Inc.3801 Regent St. Madison, Wis.53705), MUSCLE, T-Coffee, CLUSTALX, CLUSTALV, JalView, Phylip, and Discovery Studio from Accelrys (Accelrys, Inc., 10188 Telesis Ct, Suite 100, San Diego, Calif.92121). In some embodiments, an alignment will introduce “phase shifts” and / or “gaps” into one or both of the sequences being compared in order to maximize the similarity between the two sequences, and scoring refers to the process of quantitatively expressing the relatedness of the aligned sequences.

[0089] “Alpha-MF signal” or “αMF secretion signal” refers to a protein that directsnascent recombinant polypeptides to the secretory pathway.

[0090] “Arachnid” refers to a class of arthropods. For example, in someembodiments, arachnid can mean spiders, scorpions, ticks, mites, harvestmen, or solifuges.

[0091] “BAAS” means barley alpha-amylase signal peptide, and is an example of anERSP. One example of a BAAS is a BAAS having the amino acid sequence of SEQ ID NO: 37 (NCBI Accession No. AAA32925.1).

[0092] “Binary vector” or “binary expression vector” means an expression vectorwhich can replicate itself in both E. coli strains and Agrobacterium strains. Also, the vector contains a region of DNA (often referred to as t-DNA) bracketed by left and right border sequences that is recognized by virulence genes to be copied and delivered into a plant cell by Agrobacterium.

[0093] “Binding” typically refers to an association (e.g., non-covalent or covalent)between or among two or more entities (e.g., a first molecule and a second molecule or target molecule). “Direct binding” involves a direct association between two molecules, due to, e.g., covalent, electrostatic, hydrophobic, and ionic and / or hydrogen-bond interactions, including interactions such as salt bridges and water bridges; “indirect binding” involves physical interaction by way of physical contact with one or more intermediate entities. Binding between two or more entities can typically be assessed in any of a variety of contexts— including where interacting entities or moieties are studied in isolation or in the context of more complex systems (e.g., while covalently or otherwise associated with a carrier entity and / or in a biological system or cell). With regard to the binding of a first molecule to a second molecule or a target molecule, the term “specific binding” or “specifically binds to” or “binds specifically to” means binding that is measurably different from a non-specific277702-566574 interaction. Specific binding can be measured, e.g., by determining binding of a molecule compared to binding of a control molecule. For example, specific binding can be determined by competition with a control molecule that is similar to the target, for example, an excess of non-labeled target. In this case, specific binding is indicated if the binding of the labeled target to a probe is competitively inhibited by excess unlabeled target. In some embodiments, the ability of a first molecule to specifically bind to a second molecule or a target molecule can be measured either through an enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to one of skill in the art, e.g., surface plasmon resonance (SPR) technique, and traditional binding assays known by those having ordinary skill in the art. The degree of specific binding or “affinity” between a first molecule and second molecule or target molecule can be measured by common methods known in the art, including those described herein. The affinity of a first molecule to a second molecule (e.g., a target molecule) can generally be represented by the dissociation constant (“KD”). The term “KD,” as used herein, is intended to refer to the dissociation equilibrium constant of a particular compound- protein or protein-protein interaction (e.g., a first molecule and a second molecule or target molecule). In some embodiments, the affinity of a first molecule is described in terms of the KDfor an interaction between such first molecule and its target, e.g., a second molecule. For clarity, as known in the art, a smaller KD value indicates a higher affinity interaction, while a larger KDvalue indicates a lower affinity interaction. Thus, the terms “specific binding” or “specifically binds to” or is “specific for” a particular polypeptide or an epitope on a particular polypeptide target or “binds specifically to” the same, as used herein, can be exhibited, for example, by a molecule having a KD for the target of 10−4M or lower, alternatively 10−5M or lower, alternatively 10−6M or lower, alternatively 10−7M or lower, alternatively 10−8M or lower, alternatively 10−9M or lower, alternatively 10−10M or lower, alternatively 10−11M or lower, or alternatively 10−12M or even lower, as determined by, e.g., an enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to one of skill in the art, e.g., surface plasmon resonance (SPR) technique, and traditional binding assays known by those having ordinary skill in the art.

[0094] “Biomass” refers to any measured plant product.

[0095] “bp” or “base pair” refers to a molecule comprising two chemical basesbonded to one another. For example, a DNA molecule consists of two winding strands, wherein each strand has a backbone made of an alternating deoxyribose and phosphate groups. Attached to each deoxyribose is one of four bases, i.e., adenine (A), cytosine (C),277702-566574 guanine (G), or thymine (T), wherein adenine forms a base pair with thymine, and cytosine forms a base pair with guanine.

[0096] “C-terminal” refers to the free carboxyl group (i.e., -COOH) that is positionedon the terminal end of a polypeptide.

[0097] “cDNA” or “copy DNA” or “complementary DNA” refers to a molecule thatis complementary to a molecule of RNA. In some embodiments, cDNA may be either single- stranded or double-stranded. In some embodiments, cDNA can be a double-stranded DNA synthesized from a single stranded RNA template in a reaction catalyzed by a reverse transcriptase. In yet other embodiments, “cDNA” refers to all nucleic acids that share the arrangement of sequence elements found in native mature mRNA species, where sequence elements are exons and 3’ and 5’ non-coding regions. Normally, mRNA species have contiguous exons, with the intervening introns removed by nuclear RNA splicing, to create a continuous open reading frame encoding the protein. In some embodiments, “cDNA” refers to a DNA that is complementary to and derived from an mRNA template.

[0098] “CEW” refers to Corn earworm.

[0099] “Cleavable Linker” (see Linker).

[0100] “Cloning” refers to the process and / or methods concerning the insertion of aDNA segment (e.g., usually a gene of interest, for example tvp) from one source and recombining it with a DNA segment from another source (e.g., usually a vector, for example, a plasmid) and directing the recombined DNA, or “recombinant DNA” to replicate, usually by transforming the recombined DNA into a bacteria or yeast host.

[0101] “Coding sequence” or “CDS” refers to a polynucleotide or nucleic acidsequence that can be transcribed (e.g., in the case of DNA) or translated (e.g., in the case of mRNA) into a peptide, polypeptide, or protein, when placed under the control of appropriate regulatory sequences and in the presence of the necessary transcriptional and / or translational molecular factors. The boundaries of the coding sequence are determined by a translation start codon at the 5’ (amino) terminus and a translation stop codon at the 3’ (carboxy) terminus. A transcription termination sequence will usually be located 3’ to the coding sequence. In some embodiments, a coding sequence may be flanked on the 5’ and / or 3’ ends by untranslated regions. In some embodiments, a coding sequence can be used to produce a peptide, a polypeptide, or a protein product. In some embodiments, the coding sequence may or may not be fused to another coding sequence or localization signal, such as a nuclear localization signal. In some embodiments, the coding sequence may be cloned into a vector277702-566574 or expression construct, may be integrated into a genome, or may be present as a DNA fragment.

[0102] “Codon optimization” refers to the production of a gene in which one or moreendogenous, native, and / or wild-type codons are replaced with codons that ultimately still code for the same amino acid, but that are of preference in the corresponding host.

[0103] “Combination” refers to the result of combining two or more separatecomponents (e.g., a first component and one or more additional components). Thus, as used herein, a “combination” refers to an association of two or more separate components, e.g., the association of a first TVP, and one or more additional TVPs; wherein the first TVP and one or more additional TVPs are the same or different.

[0104] In some embodiments, a combination can be a “mixture.” For example, insome embodiments, a mixture refers to a combination of a first component, and one or more additional components, wherein the first component and the one or more additional components are present together in a single entity (e.g., a single unit). Thus, in some embodiments, a mixture can comprise a first component, and one or more additional components, wherein the first component and the one or more additional components are present in admixture for simultaneous administration. Accordingly, in some embodiments, a combination can refer to the association of a first TVP, and one or more additional TVPs; wherein the first TVP and one or more additional TVPs are the same or different; and wherein the first TVP and one or more additional TVPs are present in a single entity (e.g., an admixture for simultaneous administration).

[0105] In some embodiments, a combination can comprise a first component, and oneor more additional components, wherein the first component and the one or more additional components, are present separately (e.g., more than one unit). For example, in some embodiments, a combination can comprise a first component, and one or more additional components, wherein the first component and the one or more additional components may be administered separately, sequentially, simultaneously, concurrently or chronologically staggered.

[0106] Accordingly, in some embodiments, a combination can refer to the associationof a first TVP, and one or more additional TVPs; wherein the first TVP and one or more additional TVPs are the same or different; and wherein the first TVP and one or more additional TVPs are present separately (e.g., different units for separate, sequential, simultaneous, concurrent or chronologically-staggered administration).277702-566574

[0107] In some embodiments, a combination can refer to the separate, sequential,simultaneous, concurrent or chronologically-staggered application of two or more separate components (e.g., a first TVP, and one or more additional TVPs; wherein the first TVP and one or more additional TVPs are the same or different). For example, in some embodiments, a “combination” refers to the result of a simultaneous application of both a first TVP, and one or more additional TVPs; wherein the first TVP and one or more additional TVPs are the same or different. In another embodiment, a “combination” refers to the result of a separate application of a first TVP, and one or more additional TVPs; wherein the first TVP and one or more additional TVPs are the same or different.

[0108] In a further embodiments, a “combination” refers to the result of a sequentialapplication of two or more separate components, e.g., a first application of a first TVP, followed by a second application of one or more additional TVPs (wherein the first TVP and one or more additional TVPs are the same or different), or vice versa. Where the application is sequential or separate, the delay in applying the second component should not be such as to lose the beneficial effect of the combination.

[0109] “Complementary” refers to the topological compatibility or matching togetherof interacting surfaces of two polynucleotides as understood by those of skill in the art. Thus, two sequences are “complementary” to one another if they are capable of hybridizing to one another to form a stable anti-parallel, double-stranded nucleic acid structure. A first polynucleotide is complementary to a second polynucleotide if the nucleotide sequence of the first polynucleotide is substantially identical to the nucleotide sequence of the polynucleotide binding partner of the second polynucleotide, or if the first polynucleotide can hybridize to the second polynucleotide under stringent hybridization conditions. Thus, the polynucleotide whose sequence 5′-TATAC-3′ is complementary to a polynucleotide whose sequence is 5′- GTATA-3′.

[0110] “Conditioned medium” means the cell culture medium which has been usedby cells and is enriched with cell derived materials but does not contain cells.

[0111] “Copy number” refers to the number of identical copies of a vector, anexpression cassette, an amplification unit, a gene or indeed any defined nucleotide sequence, that are present in a host cell at any time. For example, in some embodiments, a gene or another defined chromosomal nucleotide sequence may be present in one, two, or more copies on the chromosome. An autonomously replicating vector may be present in one, or several hundred copies per host cell.277702-566574

[0112] “Culture” or “cell culture” refers to the maintenance of cells in an artificial, invitro environment.

[0113] “Culturing” refers to the propagation of organisms on or in various kinds ofmedia. For example, the term “culturing” can mean growing a population of cells under suitable conditions in a liquid or solid medium. In some embodiments, culturing refers to fermentative recombinant production of a heterologous polypeptide of interest and / or other desired end products (typically in a vessel or reactor).

[0114] “Cystine” refers to an oxidized cysteine-dimer. Cystines are sulfur-containingamino acids obtained via the oxidation of two cysteine molecules, and are linked with a disulfide bond.

[0115] “Defined medium” means a medium that is composed of known chemicalcomponents but does not contain crude proteinaceous extracts or by-products such as yeast extract or peptone.

[0116] “Disulfide bond” means a covalent bond between two cysteine amino acidsderived by the coupling of two thiol groups on their side chains.

[0117] “Degeneracy” or “codon degeneracy” refers to the phenomenon that oneamino acid can be encoded by different nucleotide codons. Thus, the nucleic acid sequence of a nucleic acid molecule that encodes a protein or polypeptide can vary due to degeneracies. As a result of the degeneracy of the genetic code, many nucleic acid sequences can encode a given polypeptide with a particular activity; such functionally equivalent variants are contemplated herein.

[0118] “DNA” refers to deoxyribonucleic acid, comprising a polymer of one or moredeoxyribonucleotides or nucleotides (i.e., adenine [A], guanine [G], thymine [T], or cytosine [C]), which can be arranged in single-stranded or double-stranded form. For example, one or more nucleotides creates a polynucleotide.

[0119] “dNTPs” refers to the nucleoside triphosphates that compose DNA and RNA.

[0120] “Double expression cassette” refers to two TVP expression cassettes containedon the same vector.

[0121] “Double transgene peptide expression vector” or “double transgene expressionvector” means a yeast expression vector that contains two copies of the TVP expression cassette.

[0122] “Endogenous” refers to a polynucleotide, peptide, polypeptide, protein, orprocess that naturally occurs and / or exists in an organism, e.g., a molecule or activity that is already present in the host cell before a particular genetic manipulation.277702-566574

[0123] “Enhancer element” refers to a DNA sequence operably linked to a promoter,which can exert increased transcription activity on the promoter relative to the transcription activity that results from the promoter in the absence of the enhancer element.

[0124] “ER” or “Endoplasmic reticulum” is a subcellular organelle common to alleukaryotes where some post translation modification processes occur.

[0125] “ERSP” or “endoplasmic reticulum signal peptide” is an N-terminus sequenceof amino acids that—during protein translation of the mRNA molecule encoding a TVP—is recognized and bound by a host cell signal-recognition particle, which moves the protein translation ribosome / mRNA complex to the ER in the cytoplasm. The result is the protein translation is paused until it docks with the ER where it continues and the resulting protein is injected into the ER.

[0126] “ersp” refers to a polynucleotide encoding the peptide, ERSP.

[0127] “ER trafficking” means transportation of a cell expressed protein into ER forpost-translational modification, sorting and transportation.

[0128] “Excipient” refers to any pharmacologically inactive, natural, or synthetic,component or substance that is formulated alongside (e.g., concomitantly), or subsequent to, the active ingredient of the present invention (i.e., a TVP or TVP-insecticidal protein). In some embodiments, an excipient can be any additive, adjuvant, binder, bulking agent, carrier, coating, diluent, disintegrant, filler, glidant, lubricant, preservative, vehicle, or combination thereof, with which a TVP or TVP-insecticidal protein of the present invention can be administered, and or which is useful in preparing a composition of the present invention. Excipients include any such materials known in the art that are nontoxic and do not interact with other components of a composition. In some embodiments, excipients can be formulated alongside a TVP or TVP-insecticidal protein when preparing a composition for the purpose of bulking up compositions (thus often referred to as bulking agents, fillers or diluents). In other embodiments, an excipient can be used to confer an enhancement on the active ingredient in the final dosage form, such as facilitating absorption and / or solubility. In yet other embodiments, an excipient can be used to provide stability, or prevent contamination (e.g., microbial contamination). In other embodiments, an excipient can be used to confer a physical property to a composition (e.g., a composition that is a dry granular, or dry flowable powder physical form). Reference to an excipient includes both one and more than one such excipients. Suitable pharmaceutical excipients are described in Remington's Pharmaceutical Sciences, by E.W. Martin, the disclosure of which is incorporated herein by reference in its entirety.277702-566574

[0129] “Expression cassette” refers to (1) a DNA sequence of interest, e.g., apolynucleotide operable to encode a TVP; and one or more of the following: (2) promoters, terminators, and / or enhancer elements; (3) an appropriate mRNA stabilizing polyadenylation signal; (4) an internal ribosome entry site (IRES); (5) introns; and / or (6) post-transcriptional regulatory elements. The combination (1) with at least one of (2)-(6) is called an “expression cassette.” In some embodiments, there can be numerous expression cassettes cloned into a vector. For example, in some embodiments, there can be a first expression cassettecomprising a polynucleotide operable to encode a TVP. In alternative embodiments, there aretwo expression cassettes, each comprising a polynucleotide operable to encode a TVP (i.e., a double expression cassette). In other embodiments, there are three expression cassettes operable to encode a TVP (i.e., a triple expression cassette). In some embodiments, a double expression cassette can be generated by subcloning a second expression cassette into a vector containing a first expression cassette. In some embodiments, a triple expression cassette can be generated by subcloning a third expression cassette into a vector containing a first and a second expression cassette. Methods concerning expression cassettes and cloning techniques are well-known in the art and described herein. See also TVP expression cassette.

[0130] “Expression ORF” means a nucleotide encoding a protein complex and isdefined as the nucleotides in the ORF.

[0131] “FECT” means a transient plant expression system using Foxtail mosaic viruswith elimination of coating protein gene and triple gene block.

[0132] “Fermentation beer” refers to spent fermentation medium, i.e., fermentationmedium supernatant after removal of organisms, that has been inoculated with and consumed by a transformed host cell (e.g., a yeast cell operable to express a TVP of the present invention). In some embodiments, fermentation beer refers to the solution that is recovered following the fermentation of the transformed host cell. The term “fermentation” refers broadly to the enzymatic and anaerobic or aerobic breakdown of organic substances (e.g., a carbon substrate) nutrient substances by microorganisms under controlled conditions (e.g., temperature, oxygen, pH, nutrients, and the like) to produce fermentation products (e.g., one or more peptides of the present invention). While fermentation typically describes processes that occur under anaerobic conditions, as used herein it is not intended that the term be solely limited to strict anaerobic conditions, as the term “fermentation” used herein may also occur processes that occur in the presence of oxygen.

[0133] “Fermentation solid(s)” refers to solids (including dissolved) that remain fromfermentation beer during the yeast-based fermentation process, and consists essentially of277702-566574 salts, complex protein source, vitamins, and additional yeast byproducts having a molecular weight cutoff of from about 200 kDa to about 1 kDA.

[0134] “GFP” means a green fluorescent protein from the jellyfish Aequorea victoria.

[0135] “Homologous” refers to the sequence similarity or sequence identity betweentwo polypeptides or between two nucleic acid molecules. When a position in both of the two compared sequences is occupied by the same base or amino acid monomer subunit, e.g., if a position in each of two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The percent of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared ×100. Thus, in some embodiments, the term “homologous” refers to the sequence similarity between two polypeptide molecules, or between two nucleic acid molecules. When a position in both of the two compared sequences is occupied by the same base or amino acid monomeric subunit, e.g., if a position in each of two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences. For example, if 6 of 10 of the positions in two sequences are matched or homologous then the two sequences are 60% homologous. By way of example, the DNA sequences ATTGCC and TATGGC share 50% homology. “Homology” or “homologous” or “analogous” refer to the overall relatedness between polymeric molecules, e.g., between two or more nucleic acid molecules (e.g., DNA molecules and / or RNA molecules), or between two or more polypeptide molecules.

[0136] Homology is a qualitative term that describes a relationship betweenpolymeric molecules and can be based upon the quantitative similarity or identity. Similarity or identity is a quantitative term that defines the degree of sequence match between two compared sequences. Polymeric molecules (e.g., nucleic acid molecules such as DNA molecules and / or RNA molecules, or polypeptide molecules) that share a threshold level of similarity or identity determined by alignment of matching residues are referred to herein as “homologous,” or “homologs.” Where a position in both of the two or more compared polymeric molecules is occupied by the same base or amino acid monomer subunit (e.g., if a position in each of two DNA molecules is occupied by adenine), the two or more compared sequences deemed to be homologous at that position. Thus, the homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences. For example, if half (e.g., five positions in a polymer ten subunits in length) of the positions in two sequences are homologous, the two sequences are 50% homologous; if277702-566574 90% of the positions (e.g., 9 of 10), are matched or homologous, the two sequences are 90% homologous. Accordingly, in some embodiments, the percent homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared ×100. Generally, a comparison between two sequences is made when the two sequences are aligned to give maximum homology. In some embodiments, polymeric molecules are considered to be “homologous” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical or similar. Accordingly, the term “homologous” necessarily refers to a comparison between at least two sequences (polynucleotide or polypeptide sequences).

[0137] Typically, the term “homology” implies that the compared polymericmolecules diverged in evolution from a common origin, i.e., a first polymeric molecule is related to a second polymeric molecule by descent from a common ancestral polymeric molecule; however, as used herein, it is not intended that homologous polymeric molecules (homologs) necessarily be evolutionarily related. As used herein, the terms “homology” or “homologous” or “analogous” can refer to a first polymeric molecule (or portion thereof) that exhibits similar structural or functional characteristics to a second polymeric molecule (e.g., a reference sequence), wherein the first polymeric molecule and the second polymeric molecule may or may not necessarily share a direct evolutionary relationship. Thus, as used herein, the terms “homologous” or “analogous” or “homology” can refer to the relatedness between a first polymeric molecule and a second polymeric molecule (or reference polymeric molecule)—irrespective of the evolutionary relationship between the first polymeric molecule and the second polymeric molecule. For example, in some embodiments, there may be partial homology, or complete homology and thus identical. “Sequence identity” refers to a measure of relatedness between two or more nucleic acid sequences or two or more polypeptide sequences, and is given as a percentage with reference to the total comparison length. The identity calculation takes into account those nucleotide residues or amino acid residues that are identical and in the same relative positions in their respective larger sequences. See “Identity.”

[0138] “Homologous recombination” refers to the event of substitution of a segmentof DNA by another one that possesses identical regions (homologous) or nearly so. For example, in some embodiments, “homologous recombination” refers to a type of genetic recombination in which nucleotide sequences are exchanged between two similar or identical molecules of DNA. Briefly, homologous recombination is most widely used by cells to277702-566574 accurately repair harmful breaks that occur on both strands of DNA, known as double-strand breaks. Although homologous recombination varies widely among different organisms and cell types, most forms involve the same basic steps: after a double-strand break occurs, sections of DNA around the 5′ ends of the break are cut away in a process called resection. In the strand invasion step that follows, an overhanging 3′ end of the broken DNA molecule then “invades” a similar or identical DNA molecule that is not broken. After strand invasion, the further sequence of events may follow either of two main pathways, i.e., the double- strand break repair pathway, or the synthesis-dependent strand annealing pathway. Homologous recombination is conserved across all three domains of life as well as viruses, suggesting that it is a nearly universal biological mechanism. For example, in some embodiments, homologous recombination can occur using a site-specific integration (SSI) sequence, whereby there is a strand exchange crossover event between nucleic acid sequences substantially similar in nucleotide composition. These crossover events can take place between sequences contained in the targeting construct of the invention (i.e., the SSI sequence) and endogenous genomic nucleic acid sequences (e.g., the polynucleotide encoding the peptide subunit). In addition, in some embodiments, it is possible that more than one site-specific homologous recombination event can occur, which would result in a replacement event in which nucleic acid sequences contained within the targeting construct have replaced specific sequences present within the endogenous genomic sequences.

[0139] “Hybridize” refers to the annealing of one single-stranded polynucleotide toanother polynucleotide based on the well-understood principle of sequence complementarity. In some embodiments, the other polynucleotide is a single-stranded polynucleotide. The propensity for hybridization between polynucleotides depends on the temperature and ionic strength of their milieu, the length of the polynucleotides, and the degree of complementarity. The effect of these parameters on hybridization are well known in the art.

[0140] “Hybridization” refers to any process by which a strand of polynucleotidebinds with a complementary strand through base pairing. Two single-stranded polynucleotides “hybridize” when they form a double-stranded duplex. Thus, as used herein, the term “hybridize” refers to the annealing of one single-stranded polynucleotide to another polynucleotide based on the well-understood principle of sequence complementarity. In some embodiments, the other polynucleotide is a single-stranded polynucleotide. The propensity for hybridization between polynucleotides depends on the temperature and ionic strength of their milieu, the length of the polynucleotides, and the degree of complementarity. The effect of these parameters on hybridization are well known in the art. When two single-stranded277702-566574 polynucleotides hybridize and form a double-stranded duplex, the region of double- strandedness can include the full-length of one or both of the single-stranded polynucleotides, or all of one single stranded polynucleotide and a subsequence of the other single stranded polynucleotide, or the region of double-strandedness can include a subsequence of each polynucleotide. Hybridization also includes the formation of duplexes which contain certain mismatches, provided that the two strands are still forming a double stranded helix. See “Stringent hybridization conditions” below.

[0141] “Identity” refers to a relationship between two or more polypeptide sequencesor two or more polynucleotide sequences, as determined by comparing said sequences. The term “identity” also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case may be, as determined by the match between strings of such sequences. “Identity” and “similarity” can be readily calculated by any one of the myriad methods known to those having ordinary skill in the art, including but not limited to those described in: Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994:, Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48: 1073 (1988), the disclosures of which are incorporated herein by reference in their entireties. Furthermore, methods to determine identity and similarity are codified in publicly available computer programs. For example, in some embodiments, methods to determine identity and similarity between two sequences include, but are not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(1): 387 (1984)), BLASTP, BLASTN, and FASTA (Altschul, S. F. et al., J. Molec. Biol.215: 403-410 (1990). The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBI NLM NIH Bethesda, Md. 20894; Altschul, S., et al., J. Mol. Biol.215: 403-410 (1990), the disclosures of which are incorporated herein by reference in their entireties.

[0142] “in vivo” refers to the natural environment (e.g., an animal or a cell) and toprocesses or reactions that occur within a natural environment.

[0143] “Inactive” refers to a condition wherein something is not in a state of use, e.g.,lying dormant and / or not working. For example, when used in the context of a gene or when referring to a gene, the term inactive means said gene is no longer actively synthesizing a277702-566574 gene product, having said gene product translated into a protein, or otherwise having the gene perform its normal function. For example, in some embodiments, the term inactive can refer the failure of a gene to transcribe RNA, a failure of RNA processing (e.g., pre-mRNA processing; RNA splicing; or other post-transcriptional modifications); interference with non- coding RNA maturation; interference with RNA export (e.g., from the nucleus to the cytoplasm); interference with translation; protein folding; translocation; protein transport; and / or inhibition and / or interference with any of the molecules polynucleotides, peptides, polypeptides, proteins, transcription factors, regulators, inhibitors, or other factors that take part in any of the aforementioned processes.

[0144] “Inhibiting” or “inhibit” or “combating” or “combat” or “controlling” or“control,” or any variation of these terms, refers to making something (e.g., the number of pests the functions and / or activities of the pest, and / or the deleterious effect of the pest on a plant or animal susceptible to attack thereof) less in size, amount, intensity, or degree. For example, in some embodiments, the application of a pesticidally effective amount of an TVP or agriculturally acceptable salt thereof, or an agricultural composition comprising an TVP or agriculturally acceptable salt thereof, to (i) the pest, a locus of the pest, a food supply of the pest, a habitat of the pest, or a breeding ground of the pest; (ii) a plant, a seed, a plant part, a locus of a plant, or an environment of a plant that is susceptible to an attack by the pest; (iii) an animal, a locus of an animal, or an environment of an animal susceptible to an attack by the pest; or (iv) a combination thereof, results in the following effect: a decrease in the number of pests, or inhibition of the pest’s activities (e.g., the pest dies stops or slows its movement; stops or slows its feeding; stops or slows its growth; becomes confused, e.g., with regard to navigation, locating food, sleeping behaviors, and / or mating; fails to pupate if applicable; interferes with reproduction of the pest; and / or precludes the pest from producing offspring and / or precludes the insect from producing fertile offspring) relative to the number of pests or activities thereof that had not been exposed to a pesticidally effective amount of an TVP or agriculturally acceptable salt thereof, or an agricultural composition comprising an TVP or agriculturally acceptable salt thereof.

[0145] In another embodiment, the application of a pesticidally effective amount of aTVP, TVP-pesticidal protein, or an agriculturally acceptable salt thereof, to: (i) the pest, a locus of the pest, a food supply of the pest, a habitat of the pest, or a breeding ground of the pest; (ii) a plant, a seed, a plant part, a locus of a plant, or an environment of a plant that is contacted or pollinated by a beneficial insect; (iii) an animal, a locus of an animal, or an environment of an animal susceptible to an attack by the pest; or (iv) a combination thereof,277702-566574 results in the following effect: a decrease in the number of pests, or inhibition of the pest’s activities, e.g., the pest dies stops or slows its movement; stops or slows its feeding; stops or slows its growth; becomes confused (e.g., with regard to navigation, locating food, sleeping behaviors, and / or mating); fails to pupate if applicable; interferes with reproduction and / or fecundity of the pest (e.g., interferes with or precludes the pest from producing offspring and / or interferes with or precludes the pest from producing fertile offspring); and / or weakens, destroys, or otherwise disrupts the pest’s eggs—relative to the number of pests or the level of the pest’s activities that have not been exposed to a pesticidally effective amount of a TVP, TVP-pesticidal protein, or an agriculturally acceptable salt thereof, or an agricultural composition comprising the same and one or more excipients.

[0146] In some embodiments, combating, controlling, or inhibiting a pest, includesany measurable decrease or complete inhibition to achieve a desired result. For example, there may be a decrease of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or more, in the number of pests or the activities thereof treated with peptides and / or compositions of the present disclosure, compared to untreated pests. About as used herein means within ± 10%, preferably ± 5% of a given value.

[0147] Thus, in some embodiments, the terms “combating, controlling, or inhibiting apest,” refers to a decrease in the number of pests, or an inhibition of the activities of the pests (e.g., movement; feeding; growth; level of awareness or alertness, e.g., with regard to navigation, locating food, sleeping behaviors, and / or mating; pupation if applicable; reproduction; ability to produce offspring and / or ability to produce fertile offspring) that have received a pesticidally effective amount of an TVP, TVP-pesticidal protein, or an agricultural composition comprising the same, that is at least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at least about 1%, at least about 1.25%, at least about 1.5%, at least about 1.75%, at least about 2%, at least about 2.25%, at least about 2.5%, at least about 2.75%, at least about 3%, at least about 3.25%, at least about 3.5%, at least about 3.75%, at least about 4%, at least about 4.25%, at least about 4.5%, at least about 4.75%, at least about 5%, at least about 5.25%, at least about 5.5%, at least about 5.75%, at least about 6%, at least about 6.25%, at least about 6.5%, at least about 6.75%, at least about 7%, at least about 7.25%, at least about 7.5%, at least about 7.75%, at least about 8%, at least about 8.25%, at least about 8.5%, at least about 8.75%, at least about 9%, at least about 9.25%, at least about 9.5%, at least about 9.75%, at least about 10%, at least about 11%, at least about277702-566574 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 23%, at least about 24%, at least about 25%, at least about 26%, at least about 27%, at least about 28%, at least about 29%, at least about 30%, at least about 31%, at least about 32%, at least about 33%, at least about 34%, at least about 35%, at least about 36%, at least about 37%, at least about 38%, at least about 39%, at least about 40%, at least about 41%, at least about 42%, at least about 43%, at least about 44%, at least about 45%, at least about 46%, at least about 47%, at least about 48%, at least about 49%, at least about 50%,at least about 50%, at least about 51%, at least about 52%, at least about 53%, at least about 54%, at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, or a greater than a 100%, relative to the number of pests, or the inhibition of activities of the pests (e.g., movement; feeding; growth; level of awareness or alertness, e.g., with regard to navigation, locating food, sleeping behaviors, and / or mating; pupation if applicable; reproduction; ability to produce offspring and / or ability to produce fertile offspring) that have not received a pesticidally effective amount of a TVP, TVP-pesticidal protein, or an agricultural composition thereof.

[0148] “Inoperable” refers to the condition of a thing not functioning, malfunctioning,or no longer able to function. For example, when used in the context of a gene or when referring to a gene, the term inoperable means said gene is no longer able to operate as it normally would, either permanently or transiently. For example, “inoperable,” in some embodiments, means that a gene is no longer able to synthesize a gene product, having said gene product translated into a protein, or is otherwise unable to gene perform its normal function. For example, in some embodiments, the term inoperable can refer the failure of a gene to transcribe RNA, a failure of RNA processing (e.g., pre-mRNA processing; RNA splicing; or other post-transcriptional modifications); interference with non-coding RNA maturation; interference with RNA export (e.g., from the nucleus to the cytoplasm); interference with translation; protein folding; translocation; protein transport; and / or inhibition and / or interference with any of the molecules polynucleotides, peptides,277702-566574 polypeptides, proteins, transcription factors, regulators, inhibitors, or other factors that take part in any of the aforementioned processes.

[0149] “Insect” includes all organisms in the class “Insecta.” The term “pre-adult”insects refers to any form of an organism prior to the adult stage, including, for example, eggs, larvae, and nymphs. As used herein, the term “insect refers to any arthropod and nematode, including acarids, and insects known to infest all crops, vegetables, and trees and includes insects that are considered pests in the fields of forestry, horticulture and agriculture. Examples of specific crops that might be protected with the methods disclosed herein are soybean, corn, cotton, alfalfa and the vegetable crops. A list of specific crops and insects is enclosed herein.

[0150] “Insecticidal activity” means that upon or after exposing the insect tocompounds, agents, or peptides, the insect either dies stops or slows its movement; stops or slows its feeding; stops or slows its growth; becomes confused (e.g., with regard to navigation, locating food, sleeping behaviors, and / or mating); fails to pupate; interferes with reproduction; and / or precludes the insect from producing offspring and / or precludes the insect from producing fertile offspring.

[0151] “Insect gut environment” or “gut environment” means the specific pH andproteinase conditions found within the fore, mid or hind gut of an insect or insect larva.

[0152] “Insect hemolymph environment” means the specific pH and proteinaseconditions of found within an insect or insect larva.

[0153] “Integrative expression vector” or “integrative vector” means a yeastexpression vector which can insert itself into a specific locus of the yeast cell genome and stably becomes a part of the yeast genome.

[0154] “Intervening linker” refers to a short peptide sequence in the proteinseparating different parts of the protein, or a short DNA sequence that is placed in the reading frame in the ORF to separate the upstream and downstream DNA sequences. For example, in some embodiments, an intervening linker may be used allowing proteins to achieve their independent secondary and tertiary structure formation during translation. In some embodiments, the intervening linker can be either resistant or susceptible to cleavage in plant cellular environments, in the insect and / or lepidopteran gut environment, and in the insect hemolymph and lepidopteran hemolymph environment.

[0155] “Isolated” refers to separating a thing and / or a component from its naturalenvironment, e.g., a toxin isolated from a given genus or species means that toxin is separated from its natural environment.277702-566574

[0156] “kb” refers to kilobase, i.e., 1000 bases. As used herein, the term “kb” means alength of nucleic acid molecules. For example, 1 kb refers to a nucleic acid molecule that is 1000 nucleotides long. A length of double-stranded DNA that is 1 kb long, contains two thousand nucleotides (i.e., one thousand on each strand). Alternatively, a length of single- stranded RNA that is 1 kb long, contains one thousand nucleotides.

[0157] “kDa” refers to kilodalton, a unit equaling 1,000 daltons; a “Dalton” is a unitof molecular weight (MW).

[0158] “Knock in” or “knock-in” or “knocks-in” or “knocking-in” refers to thereplacement of an endogenous gene with an exogenous or heterologous gene, or part thereof. For example, in some embodiments, the term “knock-in” refers to the introduction of a nucleic acid sequence encoding a desired protein to a target gene locus by homologous recombination, thereby causing the expression of the desired protein. In some embodiments, a “knock-in” mutation can modify a gene sequence to create a loss-of-function or gain-of- function mutation. The term “knock-in” can refer to the procedure by which a exogenous or heterologous polynucleotide sequence or fragment thereof is introduced into the genome, (e.g., “they performed a knock-in” or “they knocked-in the heterologous gene”), or the resulting cell and / or organism (e.g., “ the cell is a “knock-in” or “the animal is a “knock-in”).

[0159] “Knock out” or “knockout” or “knock-out” or “knocks-out” or “knocking-out”refers to a partial or complete suppression of the expression gene product (e.g., mRNA) of a protein encoded by an endogenous DNA sequence in a cell. In some embodiments, the “knock-out” can be effectuated by targeted deletion of a whole gene, or part of a gene encoding a peptide, polypeptide, or protein. As a result, the deletion may render a gene inactive, partially inactive, inoperable, partly inoperable, or otherwise reduce the expression of the gene or its products in any cell in the whole organism and / or cell in which it is normally expressed. The term “knock-out” can refer to the procedure by which an endogenous gene is made completely or partially inactive or inoperable (e.g., “they performed a knock-out” or “they knocked-out the endogenous gene”), or the resulting cell and / or organism (e.g., “ the cell is a “knock-out” or “the animal is a “knock-out”).

[0160] “Knockdown dose 50” or “KD50” refers to the median dose required to causeparalysis or cessation of movement in 50% of a population, for example a population of Musca domestica (common housefly) and / or Aedes aegypti (mosquito).

[0161] “l” or “linker” refers to a nucleotide encoding intervening linker peptide.277702-566574

[0162] “L” in the proper context refers to an intervening linker peptide, which links atranslational stabilizing protein (STA) with an additional polypeptide, e.g., a TVP, and / or multiple TVPs. When referring to amino acids, “L” can also mean leucine.

[0163] “LAC4 promoter” or “Lac4 promoter” refers to a DNA segment comprised ofthe promoter sequence derived from the K. lactis β-galactosidase gene. The LAC4 promoters is a strong and inducible reporter that is used to drive expression of exogenous genes transformed into yeast.

[0164] “LAC4 terminator” or “Lac4 terminator” refers to a DNA segment comprisedof the transcriptional terminator sequence derived from the K. lactis β-galactosidase gene.

[0165] “LD20” refers to a dose required to kill 20% of a population.

[0166] “LD50” refers to lethal dose 50 which means the dose required to kill 50% of apopulation.

[0167] “Lepidopteran gut environment” means the specific pH and proteinaseconditions found within the fore, mid or hind gut of a lepidopteran insect or larva.

[0168] “Lepidopteran hemolymph environment” means the specific pH andproteinase conditions of found within lepidopteran insect or larva.

[0169] “Linker” or “LINKER” or “peptide linker” or “L” or “intervening linker”refers to a short peptide sequence operable to link two peptides together. Linker can also refer to a short DNA sequence that is placed in the reading frame of an ORF to separate an upstream and downstream DNA sequences. In some embodiments, a linker can be cleavable by an insect protease. In some embodiments, a linker may allow proteins to achieve their independent secondary and tertiary structure formation during translation. In some embodiments, the linker can be either resistant or susceptible to cleavage in plant cellular environments, in the insect and / or lepidopteran gut environment, and / or in the insect hemolymph and lepidopteran hemolymph environment. In some embodiments, a linker can be cleaved by a protease, e.g., in some embodiments, a linker can be cleaved by a plant protease (e.g., papain, bromelain, ficin, actinidin, zingibain, and / or cardosins), an insect protease, a fungal protease, a vertebrate protease, an invertebrate protease, a bacteria protease, a mammal protease, a reptile protease, or an avian protease. In some embodiments, a linker can be cleavable or non-cleavable. In some embodiments, a linker comprises a binary or tertiary region, wherein each region is cleavable by at least two types of proteases: one of which is an insect and / or nematode protease and the other one of which is a human protease. In some embodiments, a linker can have one of (at least) three roles: to cleave in the insect gut environment, to cleave in the plant cell, or to be designed not to intentionally cleave.277702-566574

[0170] “Locus of a pest” refers to the habitat of a pest; food supply of a pest; breedingground of a pest; area traveled by or inhabited by a pest; material infested, eaten, used by a pest; and / or any environment in which a pest inhabits, uses, is present in, or is expected to be. In some embodiments, the locus of a pest includes, without limitation, a pest habitat; a pest food supply; a pest breeding ground; a pest area; a pest environment; any surface or location that may be frequented and / or infested by a pest; any plant or animal, or a locus of a plant or animal, susceptible to attack by a pest; and / or any surface or location where a pest may be found, may be expected to be found, or is likely to be attacked by a pest.

[0171] “Locus of a plant” refers to any place in which a plant is growing; any placewhere plant propagation materials of a plant are sown; any place where plant propagation materials of a plant will be placed into the soil; or any area where plants are stored, including without limitation, live plants and / or harvested plants, leaves, seeds, fruits, or parts thereof.

[0172] “Locus of an animal” refers to any place where animals live, eat, breed, sleep,or are otherwise present.

[0173] “Medium” (plural “media”) refers to a nutritive solution for culturing cells incell culture.

[0174] “MOA” refers to mechanism of action.

[0175] “Molecular weight (MW)” refers to the mass or weight of a molecule, and istypically measured in “daltons (Da)” or kilodaltons (kDa). In some embodiments, MW can be calculated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), analytical ultracentrifugation, or light scattering. In some embodiments, the SDS-PAGE method is as follows: the sample of interest is separated on a gel with a set of molecular weight standards. The sample is run, and the gel is then processed with a desired stain, followed by destaining for about 2 to 14 hours. The next step is to determine the relative migration distance (Rf) of the standards and protein of interest. The migration distance can be determined using the following equation: ^^^^ =Migration distance of the protein Migration distance of the dye front Formula (a)

[0176] Next, the logarithm of the MW can be determined based on the valuesobtained for the bands in the standard; e.g., in some embodiments, the logarithm of the molecular weight of an SDS-denatured polypeptide and its relative migration distance (Rf) is plotted into a graph. After plotting the graph, interpolating the value derived will provide the molecular weight of the unknown protein band.277702-566574

[0177] “Motif” refers to a polynucleotide or polypeptide sequence that is implicatedin having some biological significance and / or exerts some effect or is involved in some biological process.

[0178] “Multiple cloning site” or “MCS” refers to a segment of DNA found on avector that contains numerous restriction sites in which a DNA sequence of interest can be inserted.

[0179] “Mutant” refers to an organism, DNA sequence, amino acid sequence, peptide,polypeptide, or protein, that has an alteration or variation (for example, in the nucleotide sequence or the amino acid sequence), which causes said organism and / or sequence to be different from the naturally occurring or wild-type organism, wild-type sequence, and / or reference sequence with which the mutant is being compared. In some embodiments, this alteration or variation can be one or more nucleotide and / or amino acid substitutions or modifications (e.g., deletion or addition). In some embodiments, the one or more amino acid substitutions or modifications can be conservative; here, such a conservative amino acid substitution and / or modification in a “mutant” does not substantially diminish the activity of the mutant in relation to its non-mutant form. For example, in some embodiments, a “mutant” possesses one or more conservative amino acid substitutions when compared to a peptide with a disclosed and / or claimed sequence, as indicated by a SEQ ID NO.

[0180] “N-terminus” or “N-terminal” refers to the free amine group (i.e., -NH2) that ispositioned on beginning or start of a polypeptide.

[0181] “NCBI” refers to the National Center for Biotechnology Information.

[0182] “nm” refers to nanometers.

[0183] “Normalized peptide yield” means the peptide yield in the conditionedmedium divided by the corresponding cell density at the point the peptide yield is measured. The peptide yield can be represented by the mass of the produced peptide in a unit of volume, for example, mg per liter or mg / L, or by the UV absorbance peak area of the produced peptide in the HPLC chromatograph, for example, mAu.sec. The cell density can be represented by visible light absorbance of the culture at wavelength of 600 nm (OD600).

[0184] “OD” refers to optical density. Typically, OD is measured using aspectrophotometer. When measuring growth over time of a cell population, OD600 is preferable to UV spectroscopy; this is because at a 600 nm wavelength, the cells will not be harmed as they would under too much UV light.

[0185] “OD660nm” or “OD660nm” refers to optical densities at 660 nanometers (nm).277702-566574

[0186] “One letter code” means the peptide sequence which is listed in its one lettercode to distinguish the various amino acids in the primary structure of a protein: alanine=A, arginine=R, asparagine=N, aspartic acid=D, asparagine or aspartic acid=B, cysteine=C, glutamic acid=E, glutamine=Q, glutamine or glutamic acid=Z, glycine=G, histidine=H, isoleucine=I, leucine=L, lysine=K, methionine=M, phenylalanine=F, proline=P, serine=S, threonine=T, tryptophan=W, tyrosine=Y, and valine=V.

[0187] “Operable” refers to the ability to be used, the ability to do something, and / orthe ability to accomplish some function or result. For example, in some embodiments, “operable” refers to the ability of a polynucleotide, DNA sequence, RNA sequence, or other nucleotide sequence or gene to encode a peptide, polypeptide, and / or protein. For example, in some embodiments, a polynucleotide may be operable to encode a protein, which means that the polynucleotide contains information that imbues it with the ability to create a protein (e.g., by transcribing mRNA, which is in turn translated to protein).

[0188] “Operably linked” refers to a juxtaposition wherein the components sodescribed are in a relationship permitting them to function in their intended manner. For example, in some embodiments, operably linked can refer to two or more DNA, peptide, or polypeptide sequences. For example, in some embodiments, operably linked can mean that the two adjacent DNA sequences are placed together such that the transcriptional activation of one can act on the other. In other embodiments, the term “operably linked” can refer to peptide and / or polypeptide molecules, e.g., wherein operably linked means that two or more peptide and / or polypeptide molecules are connected in such a way as to yield a single polypeptide chain, or connected in such a way inasmuch that one peptide exerts some effect on the other. In yet other embodiments, operably linked can refer to two adjacent DNA sequences are placed together such that the transcriptional activation of one can act on the other. In other embodiments, operably linked can refer to peptide and / or polypeptide molecules, wherein two or more peptide and / or polypeptide molecules are connected in such a way as to yield a single polypeptide chain, or connected in such a way inasmuch that one peptide exerts some effect on the other.

[0189] “ORF” or “open reading frame” refers to a length of RNA or DNA sequence,between a translation start signal (e.g., AUG or ATG, respectively) and any one or more of the known termination codons, which encodes one or more polypeptide sequences. Put another way, the ORF describes the frame of reference as seen from the point of view of a ribosome translating the RNA code, insofar that the ribosome is able to keep reading (i.e., adding amino acids to the nascent protein) because it has not encountered a stop codon. Thus,277702-566574 “open reading frame” or “ORF” refers to the amino acid sequence encoded between translation initiation and termination codons of a coding sequence. Here, the terms “initiation codon” and “termination codon” refer to a unit of three adjacent nucleotides (i.e., a codon) in a coding sequence that specifies initiation and chain termination, respectively, of protein synthesis (mRNA translation).

[0190] In some embodiments, an ORF is a continuous stretch of codons that beginswith a start codon (usually ATG for DNA, and AUG for RNA) and ends at a stop codon (usually UAA, UAG or UGA). In other embodiments, an ORF can be length of RNA or DNA sequence, between a translation start signal (e.g., AUG or ATG) and any one or more of the known termination codons, wherein said length of RNA or DNA sequence encodes one or more polypeptide sequences. In some other embodiments, an ORF can be a DNA sequence encoding a protein which begins with an ATG start codon and ends with a TGA, TAA or TAG stop codon. ORF can also mean the translated protein that the DNA encodes. Generally, those having ordinary skill in the art distinguish the terms “open reading frame” and “ORF,” from the term “coding sequence,” based upon the fact that the broadest definition of “open reading frame” simply contemplates a series of codons that does not contain a stop codon. Accordingly, while an ORF may contain introns, the coding sequence is distinguished by referring to those nucleotides (e.g., concatenated exons) that can be divided into codons that are actually translated into amino acids by the ribosomal translation machinery (i.e., a coding sequence does not contain introns); however, as used herein, the terms “coding sequence”; “CDS”; “open reading frame”; and “ORF,’ are used interchangeably.

[0191] “Out-recombined” or “out-recombination” refers to the removal of a geneand / or polynucleotide sequence (e.g., an endogenous gene) that is flanked by two site- specific recombination sites (e.g., the 5’- and 3’- nucleotide sequence of a target gene that is homologous to the homology arms of a target vector) during in vivo homologous recombination. See “knockout.”

[0192] “Pest” refers to any organism (e.g., including, but is not limited to: insects,fungi, bacteria, nematodes, mites, ticks, snails, and the like) that is annoying, troublesome, detrimental, harmful, a nuisance, attacks, is directly or indirectly harmful, and / or causes damage or destruction, for example, to people, plants, crops, pets, a beneficial insect, and / or livestock.

[0193] In some embodiments, a pest can be an organism (e.g., an arthropod, such asan insect) that directly or indirectly harms the plant. In some embodiments, a pest can have a harmful direct effect on plant. For example, in some embodiments, a pest can be insect (e.g.,277702-566574 a phytophagous insect pest) that causes a harmful direct effect on a plant by feeding on the plant leaves.

[0194] In yet other embodiments, a pest can be an insect (e.g., a phytopathogenicinsect) that causes a harmful indirect effect on a plant, e.g., via transmission of a disease agent (e.g., a virus, bacteria, etc.) from the insect to the plant. Accordingly, in some embodiments, the pest serves as a vector for pathogen transmission.

[0195] In some embodiments, a pest can be a phytophagous insect that consumes,eats, or otherwise injures a seed or plant grown therefrom, or any part of a plant, e.g., plant tissues, plant cells, plant parts, plant organs (e.g., leaves, stems, roots, etc.), seeds, propagules, embryos and progeny of the same.

[0196] “Pesticidal” is generally used to refer to the ability of a peptide of the presentdisclosure, or a composition comprising the same, to increase mortality, inhibit the growth rate, or interfere with one or more activities, of a pest, e.g., a deleterious arthropod insect. As used herein, “insecticidal” is generally used to refer to the ability of a peptide of the present disclosure, or a composition comprising the same, to increase mortality, inhibit the growth rate, or interfere with one or more activities, of an insect.

[0197] “Pesticidal activity” or “insecticidal activity” (e.g., in the context of insectpests) means that upon or after exposing the pest to a peptide of the present disclosure or a composition comprising a peptide of the present disclosure and an excipient, the pest (e.g., a deleterious insect) either dies, stops or slows its movement; stops or slows its feeding; stops or slows its growth; becomes confused (e.g., with regard to navigation, locating food, sleeping behaviors, and / or mating); fails to pupate; interferes with reproduction; and / or precludes the insect from producing offspring and / or precludes the insect from producing fertile offspring, or any combination of the foregoing.

[0198] “Pesticidal activity assay” or “insecticidal activity assay” (in the context ofinsect pests) refers to an assay wherein one or more pests (e.g., one or more deleterious insects) are treated with either an agent (e.g., a TVP or an agriculturally acceptable salt thereof, a TVP-insecticidal protein or an agriculturally acceptable salt thereof, or a composition comprising the same), or a comparator or control, and wherein the insecticidal activity of the agent or the comparator or control is then determined 12-, 18-, or 24-hours post-treatment based on the dose (ng / µL) of the agent, or the dose (ng / µL) of the comparator or control, which is required to achieve 100% insect mortality, i.e., 100% of insects are dead, unmoving, or unable to hold on to the side of a container (or host in the context of a parasite) when said container (or host) is up-ended.277702-566574

[0199] In some embodiments, the pesticidal activity of the agent relative to thepesticidal activity the comparator or control can be compared, wherein (1) at least a first group of one or more pests (e.g., insect pests) is treated with the agent (e.g., a TVP or an agriculturally acceptable salt thereof, a TVP-insecticidal protein or an agriculturally acceptable salt thereof, or a composition comprising the same); and (2) at least a second group of one or more pests (e.g., insect pests) is treated with the comparator or control; and wherein the pesticidal activity (in this example, the insecticidal activity) of the agent relative to the comparator or control is then determined, e.g., 24-hours post-treatment, based on the dose (ng / µL) of the agent, relative to the dose (ng / µL) of the comparator or control, required to achieve 100% insect mortality (i.e., 100% of the insects are dead, unmoving, or unable to hold on to the side of a container or host, when said container or host is up-ended).

[0200] Accordingly, where an agent such as the peptides and / or the compositions ofthe present disclosure has an pesticidal activity that is greater than a comparator or control, e.g., a comparator such as a vehicle, water, etc.—as measured via a pesticidal activity assay as described herein—the dose (ng / µL) of the agent required to achieve 100% pest mortality (i.e., 100% of the one or more pests are dead, unmoving, or unable to hold on to the side of a container or host when said container or host is up-ended) will be less than the dose (ng / µL) of the comparator or control required to achieve 100% pest mortality.

[0201] For example, in some embodiments, a pesticidal activity assay can beperformed as follows: one or more pests (e.g., insect pests) can be immobilized via CO2 for 10 minutes, and then transferred to a CO2pad to keep them immobilized. Insects of a predetermined size (e.g., weight in mg) can be picked for topical application of the agent or control. A dose concentration of an agent (e.g., a TVP or an agriculturally acceptable salt thereof, a TVP-insecticidal protein or an agriculturally acceptable salt thereof, or a composition comprising the same) and a comparator (e.g., water, vehicle, or a untreated control) can be estimated by peak area using reverse phase HPLC. Next, the treatment can be applied, e.g., 0.5 µL of a solution containing the agent, or a control solution containing water, can be topically applied to each insect, or the environment (e.g., host environment). Here, the solution containing the agent is topically applied to insects in a first group, and the control solution containing water is topically applied to insects in a second group. The treated insects and untreated insects can then each be transferred to their own container, which may optionally contain a host organism (e.g., to determine whether the agent has an effect on the host). “Insect mortality,” i.e., insect knock-down and insect death, can then be assessed at 24- hours post-treatment of the single dose; here, “insect mortality” is the dose (ng / µL) required277702-566574 for 100% of the insects to be either dead, unmoving, or unable to hold on to the side of a container or host when said container or host is up-ended.

[0202] “Pesticidal effect” or “insecticidal effect” (e.g., in the context of insects) refersto the removal or the reduction of harm of pests (e.g., deleterious insect pests). The concept of “insecticidal effect” includes reducing of the target insect, killing of insects (extermination), insect proliferation inhibition, insect development inhibition, insect growth inhibition, repelling of insects (repellence), reducing of the survival rate of the target insect and the removal or the reduction of harm of insects (for example, inhibition of ingestion capacity of agricultural insects). “Insecticidal effect” includes killing of any individual or group of insects.

[0203] “Pesticidally-effective amount” or “insecticidally-effective amount” refers toan amount of (1) a TVP, or TVP-insecticidal protein of the present disclosure, or an agriculturally acceptable salt thereof; and / or (2) an insecticidal composition comprising: a TVP, or TVP-insecticidal protein of the present disclosure, or an agriculturally acceptable salt thereof, and an excipient; that is sufficient to: partially or completely inhibit an pest (e.g., a deleterious arthropod insect); bring about the death of at least one pest; partially or completely reduce or decrease insect growth, feeding, or normal physiological development; partially or completely inhibit or decrease the normal insect cellular processes, including maintenance and growth; and / or attenuate or decrease the severity of an insect infestation. This amount will vary depending on such factors including but not limited to: the specific target insect to be controlled; the specific environment, location, plant, crop, or agricultural site to be treated; the environmental conditions, method, rate, concentration, stability, and quantity applied. Further, those having ordinary skill in the art will recognize that the insecticidally-effective amount may also vary with respect to climatic conditions, environmental considerations, and / or frequency of application and / or severity of insect infestation. In some embodiments, insecticidally-effective amounts can be measured by the use of assays that measure the reduction in growth or decline in insect populations. One measure of reduction can be to express the decrease in population in logarithmic scale typical of a specific microbial species. That is, a 1 log reduction is equivalent to a 90% reduction versus a control, a 2 log reduction is a 99% reduction, etc.

[0204] “Pharmaceutically acceptable salt” is synonymous with agriculturallyacceptable salt, and as used herein refers to a compound that is modified by making acid or base salts thereof.277702-566574

[0205] “Plant” shall mean whole plants, plant tissues, plant organs (e.g., leaves,stems, roots, etc.), seeds, plant cells, propagules, embryos and progeny of the same. Plant cells can be differentiated or undifferentiated (e.g. callus, suspension culture cells, protoplasts, leaf cells, root cells, phloem cells, and pollen).

[0206] “Plant transgenic protein” means a protein from a heterologous species that isexpressed in a plant after the DNA or RNA encoding it was delivered into one or more of the plant cells.

[0207] “Plasmid” refers to a DNA segment that acts as a carrier for a gene of interest(e.g., tvp) and, when transformed or transfected into an organism, can replicate and express the DNA sequence contained within the plasmid independently of the host organism. Plasmids are a type of vector, and can be “cloning vectors” (i.e., simple plasmids used to clone a DNA fragment and / or select a host population carrying the plasmid via some selection indicator) or “expression plasmids” (i.e., plasmids used to produce large amounts of polynucleotides and / or polypeptides).

[0208] “Polynucleotide” refers to a polymeric-form of nucleotides (e.g.,ribonucleotides, deoxyribonucleotides, or analogs thereof) of any length; e.g., a sequence of two or more ribonucleotides or deoxyribonucleotides. As used herein, the term “polynucleotide” includes double- and single-stranded DNA, as well as double- and single- stranded RNA; it also includes modified and unmodified forms of a polynucleotide (modifications to and of a polynucleotide, for example, can include methylation, phosphorylation, and / or capping). In some embodiments, a polynucleotide can be one of the following: a gene or gene fragment (for example, a probe, primer, EST, or SAGE tag); genomic DNA; genomic DNA fragment; exon; intron; messenger RNA (mRNA); transfer RNA; ribosomal RNA; ribozyme; cDNA; recombinant polynucleotide; branched polynucleotide; plasmid; vector; isolated DNA of any sequence; isolated RNA of any sequence; nucleic acid probe; primer or amplified copy of any of the foregoing.

[0209] In yet other embodiments, a polynucleotide can refer to a polymeric-form ofnucleotides operable to encode the open reading frame of a gene.

[0210] In some embodiments, a polynucleotide can refer to cDNA.

[0211] In some embodiments, polynucleotides can have any three-dimensionalstructure and may perform any function, known or unknown. The structure of a polynucleotide can also be referenced to by its 5’- or 3’- end or terminus, which indicates the directionality of the polynucleotide. Adjacent nucleotides in a single-strand of polynucleotides are typically joined by a phosphodiester bond between their 3’ and 5’277702-566574 carbons. However, different internucleotide linkages could also be used, such as linkages that include a methylene, phosphoramidate linkages, etc. This means that the respective 5’ and 3’ carbons can be exposed at either end of the polynucleotide, which may be called the 5’ and 3’ ends or termini. The 5’ and 3’ ends can also be called the phosphoryl (PO4) and hydroxyl (OH) ends, respectively, because of the chemical groups attached to those ends. The term polynucleotide also refers to both double- and single-stranded molecules. Unless otherwise specified or required, any embodiment that makes or uses a polynucleotide encompasses both the double-stranded form and each of two complementary single-stranded forms known or predicted to make up the double-stranded form.

[0212] In some embodiments, a polynucleotide can include modified nucleotides,such as methylated nucleotides and nucleotide analogs (including nucleotides with non- natural bases, nucleotides with modified natural bases such as aza- or deaza-purines, etc.). If present, modifications to the nucleotide structure can be imparted before or after assembly of the polynucleotide.

[0213] In some embodiments, a polynucleotide can also be further modified afterpolymerization, such as by conjugation with a labeling component. Additionally, the sequence of nucleotides in a polynucleotide can be interrupted by non-nucleotide components. One or more ends of the polynucleotide can be protected or otherwise modified to prevent that end from interacting in a particular way (e.g. forming a covalent bond) with other polynucleotides.

[0214] In some embodiments, a polynucleotide can be composed of a specificsequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T). Uracil (U) can also be present, for example, as a natural replacement for thymine when the polynucleotide is RNA. Uracil can also be used in DNA. Thus, the term “sequence” refers to the alphabetical representation of a polynucleotide or any nucleic acid molecule, including natural and non-natural bases.

[0215] The term “RNA molecule” or ribonucleic acid molecule refers to apolynucleotide having a ribose sugar rather than deoxyribose sugar and typically uracil rather than thymine as one of the pyrimidine bases. An RNA molecule of the invention is generally single-stranded, but can also be double-stranded. In the context of an RNA molecule from an RNA sample, the RNA molecule can include the single-stranded molecules transcribed from DNA in the cell nucleus, mitochondrion or chloroplast, which have a linear sequence of nucleotide bases that is complementary to the DNA strand from which it is transcribed.277702-566574

[0216] In some embodiments, a polynucleotide can further comprise one or moreheterologous regulatory elements. For example, in some embodiments, the regulatory element is one or more promoters; enhancers; silencers; operators; splicing signals; polyadenylation signals; termination signals; RNA export elements, internal ribosomal entry sites (IRES); poly-U sequences; or combinations thereof.

[0217] “Post-transcriptional regulatory elements” are DNA segments and / ormechanisms that affect mRNA after it has been transcribed. Mechanisms of post- transcriptional mechanisms include splicing events; capping, splicing, and addition of a Poly (A) tail, and other mechanisms known to those having ordinary skill in the art.

[0218] “Promoter” refers to a region of DNA to which RNA polymerase binds andinitiates the transcription of a gene.

[0219] “Protein” and “polypeptide” and “peptide” are used interchangeably herein torefer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical analogue of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers. The terms “protein” and “polypeptide” and “peptide” are also inclusive of modifications including, but not limited to, glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-ribosylation.

[0220] “Ratio” refers to the quantitative relation between two amounts showing thenumber of times one value contains or is contained within the other.

[0221] “Reading frame” refers to one of the six possible reading frames, three in eachdirection, of the double stranded DNA molecule. The reading frame that is used determines which codons are used to encode amino acids within the coding sequence of a DNA molecule. In some embodiments, a reading frame is a way of dividing the sequence of nucleotides in a polynucleotide and / or nucleic acid (e.g., DNA or RNA) into a set of consecutive, non-overlapping triplets.

[0222] “Recombinant DNA” or “rDNA” refers to DNA that is comprised of two ormore different DNA segments.

[0223] “Recombinant vector” means a DNA plasmid vector into which foreign DNAhas been inserted.

[0224] “Regulatory elements” refers to a genetic element that controls some aspect ofthe expression and / or processing of nucleic acid sequences. For example, in some embodiments, a regulatory element can be found at the transcriptional and post-transcriptional level. Regulatory elements can be cis-regulatory elements (CREs), or trans-277702-566574 regulatory elements (TREs). In some embodiments, a regulatory element can be one or more promoters; enhancers; silencers; operators; splicing signals; polyadenylation signals; termination signals; RNA export elements, internal ribosomal entry sites (IRES); poly-U sequences; and / or other elements that influence gene expression, for example, in a tissue- specific manner; temporal-dependent manner; to increase or decrease expression; and / or to cause constitutive expression.

[0225] “Restriction enzyme” or “restriction endonuclease” refers to an enzyme thatcleaves DNA at a specified restriction site. For example, a restriction enzyme can cleave a plasmid at an EcoRI, SacII or BstXI restriction site allowing the plasmid to be linearized, and the DNA of interest to be ligated.

[0226] “Restriction site” refers to a location on DNA comprising a sequence of 4 to 8nucleotides, and whose sequence is recognized by a particular restriction enzyme.

[0227] “Selection gene” means a gene which confers an advantage for a geneticallymodified organism to grow under the selective pressure.

[0228] “Serovar” or “serotype” refers to a group of closely related microorganismsdistinguished by a characteristic set of antigens. In some embodiments, a serovar is an antigenically and serologically distinct variety of microorganism

[0229] “sp.” refers to species.

[0230] “ssp.” or “subsp.” refers to subspecies.

[0231] “Subcloning” or “subcloned” refers to the process of transferring DNA fromone vector to another, usually advantageous vector. For example, polynucleotide encoding a mutant TVP can be subcloned into a pKlac1 plasmid subsequent to selection of yeast colonies transformed with pKLAC1 plasmids.

[0232] “SSI” is an acronym that is context dependent. In some contexts, it can refer to“Site-specific integration,” which is used to refer to a sequence that will permit in vivo homologous recombination to occur. However, in other contexts, SSI can refer to “Surface spraying indoors,” which is a technique of applying a variable volume sprayable volume of an insecticide onto indoor surfaces where vectors rest, such as on walls, windows, floors and ceilings. The term “site-specific integration” refers to the process directing a transgene to a target site in a host-organism’s genome; thus, SSI allows the integration of genes of interest into pre-selected genome locations of a host-organism.

[0233] “STA” or “Translational stabilizing protein” or “stabilizing domain” or“stabilizing protein” (used interchangeably herein) means a peptide or protein with sufficient tertiary structure that it can accumulate in a cell without being targeted by the cellular process277702-566574 of protein degradation. The protein can be between 5 and 50 amino acids long. The translational stabilizing protein is coded by a DNA sequence for a protein that is operably linked with a sequence encoding an insecticidal protein or a TVP in the ORF. The operably- linked STA can either be upstream or downstream of the TVP and can have any intervening sequence between the two sequences (STA and TVP) as long as the intervening sequence does not result in a frame shift of either DNA sequence. The translational stabilizing protein can also have an activity which increases delivery of the TVP across the gut wall and into the hemolymph of the insect.

[0234] “sta” means a nucleotide encoding a translational stabilizing protein.

[0235] “Stringent hybridization” or “stringent hybridization conditions” refers toconditions under which a polynucleotide (e.g., a nucleic acid probe, primer or oligonucleotide) will hybridize to its target sequence, typically in a complex mixture of nucleic acids, but not to other sequences. In some embodiments, the term “stringent hybridization” or “stringent hybridization conditions” refers to the conditions under which a probe will hybridize to its target sequence to a detectably greater degree than to other sequences (e.g., at least 2-fold, 5-fold, or 10-fold over background).

[0236] Stringent hybridization conditions are sequence- and length-dependent, andwill be different in different circumstances. Similarly, stringent hybridization conditions depend on % (percent)-identity (or %-mismatch) over a certain length of nucleotide residues. Generally, longer sequences hybridize specifically at higher temperatures than shorter sequences. By controlling the stringency of the hybridization and / or washing conditions, target sequences that are 100% complementary to the probe can be identified (homologous probing). Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity are detected (heterologous probing). Generally, a probe is less than about 1000 or 500 nucleotides in length

[0237] For example, in some embodiments, stringent hybridization conditions will bethose in which the salt concentration is less than about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes (e.g., 10 to 50 nucleotides), and at least about 60° C. for long probes (e.g., greater than 50 nucleotides). In some embodiments, stringent hybridization conditions may also be achieved with the addition of destabilizing agents such as formamide.

[0238] In some embodiments, low stringency hybridization conditions includehybridization with a buffer solution of 30 to 35% formamide, 1 M NaCl, 1% SDS (sodium dodecyl sulfate) at 37°C, and a wash in 1× to 2×SSC (20×SSC=3.0 M NaCl / 0.3 M trisodium277702-566574 citrate) at 50-55°C. In some embodiments, moderate stringency hybridization conditions include hybridization in 40 to 45% formamide, 1.0 M NaCl, 1% SDS at 37°C, and a wash in 0.5× to 1×SSC at 55-60°C. In some embodiments, high stringency hybridization conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37°C., and a final wash in 0.1×SSC at 60 to 65°C. for at least about 20 minutes. Optionally, wash buffers may comprise about 0.1% to about 1% SDS. The duration of hybridization is generally less than about 24 hours, usually about 4 to about 12 hours.

[0239] Specificity is typically the function of post-hybridization washes, the criticalfactors being the ionic strength and temperature of the final wash solution. For DNA-DNA hybrids, the Tm(thermal melting point) can be approximated from the equation of Meinkoth and Wahl (1984) Anal. Biochem.138:267-284: Tm=81.5° C.+16.6 (log M)+0.41 (% GC)−0.61 (% form)−500 / L; where M is the molarity of monovalent cations, % GC is the percentage of guanosine and cytosine nucleotides in the DNA, “% form” is the percentage of formamide in the hybridization solution, and L is the length of the hybrid in base pairs. The Tm is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. Washes are typically performed at least until equilibrium is reached and a low background level of hybridization is achieved, such as for 2 hours, 1 hour, or 30 minutes.

[0240] In some embodiments, the Tm is reduced by about 1°C for each 1% ofmismatching; thus, Tm, hybridization, and / or wash conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences with >90% identity are sought, the Tm can be decreased 10°C.

[0241] In some embodiments, stringent hybridization conditions are selected to beabout 5°C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. However, in some embodiments, stringent hybridization conditions can utilize a hybridization and / or wash temperature that is about 1°C, 2°C, 3°C, or 4°C lower than the thermal melting point (Tm).

[0242] Using the equation, hybridization and wash compositions, and desired Tm,those of ordinary skill will understand that variations in the stringency of hybridization and / or wash solutions are inherently described. An exemplary description of the hybridization of nucleic acids is found in Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology—Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, New York); and Ausubel et al., eds. (1995) Current Protocols in Molecular Biology, Chapter277702-566574 2 (Greene Publishing and Wiley-Interscience, New York); the disclosures of which are incorporated herein by reference in their entireties.

[0243] In some embodiments, a polynucleotide of the present disclosure canstringently hybridize to a polynucleotide encoding a mannose-binding lectin, or a complementary nucleotide sequence thereof.

[0244] “Structural motif” refers to the three-dimensional arrangement of peptidesand / or polypeptides, and / or the arrangement of operably linked polypeptide segments. For example, a polypeptide having an ERSP motif, an STA motif, a LINKER motif, and a TVP polypeptide motif, has an overall “structural motif” of ERSP-STA-L-TVP. See also “TVP construct.”

[0245] “Susceptible to attack by a pest(s),” refer to plants, or human or animalpatients or subjects, susceptible to a pest or a pest infections.

[0246] “Ta1b” or “U1-agatoxin-Ta1b” or “Ta1bWT” or “wild-type U1-agatoxin-Ta1b” refers to a polypeptide isolated from the Hobo spider, Eratigena agrestis. One example of a U1-agatoxin-Ta1b is a polypeptide having the amino acid sequence of SEQ ID NO: 1 (NCBI Accession No. O46167.1).

[0247] “Ta1b variant polynucleotide” or “U1-agatoxin-Ta1b variant polynucleotide”refers to a polynucleotide or group of polynucleotides operable to express and / or encode an insecticidal protein comprising one or more TVPs. The term “U1-agatoxin-Ta1b variant polynucleotide” when used to describe the U1-agatoxin-Ta1b variant polynucleotide sequence contained in a TVP expression ORF, its inclusion in a vector, and / or when describing the polynucleotides encoding an insecticidal protein, is described as “tvp” and / or “Tvp.”

[0248] “Toxin” refers to a venom and / or a poison, especially a protein or conjugatedprotein produced by certain animals, higher plants, and pathogenic bacteria. Generally, the term “toxin” is reserved natural products, e.g., molecules and peptides found in scorpions, spiders, snakes, poisonous mushrooms, etc., whereas the term “toxicant” is reserved for man- made products and / or artificial products e.g., man-made chemical pesticides. However, as used herein, the terms “toxin” and “toxicant” are used synonymously

[0249] “Transfection” and “transformation” both refer to the process of introducingexogenous and / or heterologous DNA or RNA (e.g., a vector containing a polynucleotide that encodes a TVP) into a host organism (e.g., a prokaryote or a eukaryote). Generally, those having ordinary skill in the art sometimes reserve the term “transformation” to describe processes where exogenous and / or heterologous DNA or RNA are introduced into a bacterial277702-566574 cell; and reserve the term “transfection” for processes that describe the introduction of exogenous and / or heterologous DNA or RNA into eukaryotic cells. However, as used herein, the term “transformation” and “transfection” are used synonymously, regardless of whether a process describes the introduction exogenous and / or heterologous DNA or RNA into a prokaryote (e.g., bacteria) or a eukaryote (e.g., yeast, plants, or animals).

[0250] “Transgene” means a heterologous DNA sequence encoding a protein which istransformed into a plant.

[0251] “Transgenic host cell” means a cell which is transformed with a gene and hasbeen selected for its transgenic status via an additional selection gene.

[0252] “Transgenic plant” means a plant that has been derived from a single cell thatwas transformed with foreign DNA such that every cell in the plant contains that transgene.

[0253] “Transient expression system” means an Agrobacterium tumefaciens-basedsystem which delivers DNA encoding a disarmed plant virus into a plant cell where it is expressed. The plant virus has been engineered to express a protein of interest at high concentrations, up to 40% of the TSP.

[0254] “Triple expression cassette refers to three TVP expression cassettes containedon the same vector.

[0255] “TRBO” means a transient plant expression system using Tobacco mosaicvirus with removal of the viral coating protein gene.

[0256] “TSP” or “total soluble protein” means the total amount of protein that can beextracted from a plant tissue sample and solubilized into the extraction buffer.

[0257] “TVP” or “U1-agatoxin-Ta1b Variant Polypeptides (TVPs)” or “Ta1b VariantPolypeptides (TVPs)” refers to mutants or variants of the wild-type U1-agatoxin-Ta1b polypeptide sequence and / or a polynucleotide sequence encoding a wild-type U1-agatoxin- Ta1b polypeptide, that have been altered to produce a non-naturally occurring polypeptide and / or polynucleotide sequence. An exemplary wild-type U1-agatoxin-Ta1b polypeptide sequence is provided herein, having the amino acid sequence of SEQ ID NO: 1. A TVP polypeptide includes peptides that range in size from about 41 contiguous amino acids to about 51 contiguous amino acids, and every integer therebetween, for example, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, and 51 contiguous amino acids.

[0258] “TVP ORF diagram” refers to the composition of one or more TVP ORFs, aswritten out in diagram or equation form. For example, a “TVP ORF diagram” can be written out as using acronyms or short-hand references to the DNA segments contained within the expression ORF. Accordingly, in one example, a “TVP ORF diagram” may describe the277702-566574 polynucleotide segments encoding the ERSP, LINKER, STA, and TVP, by diagramming in equation form the DNA segments as “ersp” (i.e., the polynucleotide sequence that encodes the ERSP polypeptide); “linker” or “L” (i.e., the polynucleotide sequence that encodes the LINKER polypeptide); “sta” (i.e., the polynucleotide sequence that encodes the STA polypeptide), and “tvp” (i.e., the polynucleotide sequence encoding a TVP), respectively. An example of a TVP ORF diagram is “ersp-sta-(linkeri-tvpj)N,” or “ersp-(tvpj-linkeri)N-sta” and / or any combination of the DNA segments thereof.

[0259] “TVP polynucleotide” refers to a polynucleotide or group of polynucleotidesoperable to express and / or encode a TVP or a TVP-insecticidal protein.

[0260] “TVP-insecticidal protein” refers to any protein, peptide, polypeptide, aminoacid sequence, configuration, or arrangement, consisting of: (1) at least one TVP, or two or more TVPs; and (2) additional non-toxin peptides, polypeptides, or proteins, wherein said additional non-toxin peptides, polypeptides, or proteins e.g., in some embodiments, have the ability to do one or more of the following: increase the mortality and / or inhibit the growth of insects when the insects are exposed to a TVP-insecticidal protein, relative to a TVP alone; increase the expression of said TVP-insecticidal protein, e.g., in a host cell or an expression system; and / or affect the post-translational processing of the TVP-insecticidal protein. In some embodiments, a TVP-insecticidal protein can be a polymer comprising two or more TVPs. In some embodiments, a TVP-insecticidal protein can be a polymer comprising two or more TVPs, wherein the TVPs are operably linked via a linker peptide, e.g., a cleavable and / or non-cleavable linker. In some embodiments, a TVP-insecticidal protein can refer to a one or more TVPs operably linked with one or more proteins such as a stabilizing domain (STA); an endoplasmic reticulum signaling protein (ERSP); an insect cleavable or insect non- cleavable linker (L); and / or any other combination thereof. In some embodiments, a TVP- insecticidal protein can be a non-naturally occurring protein comprising (1) a wild-type Ta1b protein; and (2) additional non-toxin peptides, polypeptides, or proteins, e.g., an ERSP; a linker; a STA; a UBI; or a histidine tag or similar marker.

[0261] “TVP construct” refers to the three-dimensional arrangement / orientation ofpeptides, polypeptides, and / or motifs of operably linked polypeptide segments (e.g., a TVP- insecticidal protein). For example, a TVP expression ORF can include one or more of the following components or motifs: a TVP; an endoplasmic reticulum signal peptide (ERSP); a linker peptide (L); a translational stabilizing protein (STA); or any combination thereof. And, as used herein, the term “TVP construct” is used to describe the designation and / or orientation of the structural motif. In other words, the TVP construct describes the277702-566574 arrangement and orientation of the components or motifs contained within a given TVP expression ORF. For example, in some embodiments, a TVP construct describes, without limitation, the orientation of one of the following TVP-insecticidal proteins: ERSP-TVP; ERSP-(TVP)N; ERSP-TVP-L; ERSP-(TVP)N-L; ERSP-(TVP-L)N; ERSP-L-TVP; ERSP-L- (TVP)N; ERSP-(L-TVP)N; ERSP-STA-TVP; ERSP-STA-(TVP)N; ERSP-TVP-STA; ERSP- (TVP)N-STA; ERSP-(STA-TVP)N; ERSP-(TVP-STA)N; ERSP-L-TVP-STA; ERSP-L-STA- TVP; ERSP-L-(TVP-STA)N; ERSP-L-(STA-TVP)N; ERSP-L-(TVP)N-STA; ERSP-(L- TVP)N-STA; ERSP-(L-STA-TVP)N; ERSP-(L-TVP-STA)N; ERSP-(L-STA)N-TVP; ERSP- (L-TVP)N-STA; ERSP-STA-L-TVP; ERSP-STA-TVP-L; ERSP-STA-L-(TVP)N; ERSP- (STA-L)N-TVP; ERSP-STA-(L-TVP)N; ERSP-(STA-L-TVP)N; ERSP-STA-(TVP)N-L; ERSP-STA-(TVP-L)N; ERSP-(STA-TVP)N-L; ERSP-(STA-TVP-L)N; ERSP-TVP-L-STA; ERSP-TVP-STA-L; ERSP-(TVP)N-STA-L ERSP-(TVP-L)N-STA; ERSP-(TVP-STA)N-L; ERSP-(TVP-L-STA)N; or ERSP-(TVP-STA-L)N; wherein N is an integer ranging from 1 to 200. See also “Structural motif.”

[0262] “var.” refers to varietas or variety. The term “var.” is used to indicate ataxonomic category that ranks below the species level and / or subspecies (where present). In some embodiments, the term “var.” represents members differing from others of the same subspecies or species in minor but permanent or heritable characteristics.

[0263] “Variant” or “variant sequence” or “variant peptide” refers to an amino acidsequence that possesses one or more conservative amino acid substitutions or conservative modifications. The conservative amino acid substitutions in a “variant” does not substantially diminish the activity of the variant in relation to its non-varied form. For example, in some embodiments, a “variant” possesses one or more conservative amino acid substitutions when compared to a peptide with a disclosed and / or claimed sequence, as indicated by a SEQ ID NO.

[0264] “Vector” refers to the DNA segment that accepts a foreign gene of interest(e.g., tvp). The gene of interest is known as an “insert” or “transgene.”

[0265] “Vitrification” refers to a process of converting a material into a glass-likeamorphous material. The glass-like amorphous solid may be free of any crystalline structure. Solidification of a vitreous solid occurs at the glass transition temperature (Tg).

[0266] “Wild type” or “WT” or “wild-type” or “wildtype” or “native” (all usedinterchangeably) refer to the phenotype and / or genotype (i.e., the appearance or sequence) of an organism, polynucleotide sequence, and / or polypeptide sequence, as it is found and / or observed in its naturally occurring state or condition (e.g., in the wild or nature).277702-566574

[0267] “Yeast expression vector” or “expression vector” or “vector” means a plasmidwhich can introduce a heterologous gene and / or expression cassette into yeast cells to be transcribed and translated.

[0268] “Yield” refers to the production of a peptide, and increased yields can meanincreased amounts of production, increased rates of production, and an increased average or median yield and increased frequency at higher yields. The term “yield” when used in reference to plant crop growth and / or production, as in “yield of the plant” refers to the quality and / or quantity of biomass produced by the plant.

[0269] The terms “first,” “second,” and the like, herein do not denote any order,quantity, or importance, but rather are used to distinguish one element from another, and the terms “a” and “an” herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. All ranges disclosed herein are inclusive and combinable.

[0270] Throughout this specification, unless specifically stated otherwise or thecontext requires otherwise, reference to a single step, composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e., one or more) of those steps, compositions of matter, groups of steps or group of compositions of matter.

[0271] The present disclosure is performed without undue experimentation using,unless otherwise indicated, conventional techniques of molecular biology, microbiology, virology, recombinant DNA technology, solid phase and liquid nucleic acid synthesis, peptide synthesis in solution, solid phase peptide synthesis, immunology, cell culture, and formulation. Such procedures are described, for example, in Sambrook, Fritsch & Maniatis, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, New York, Second Edition (1989), whole of Vols I, II, and III; DNA Cloning: A Practical Approach, Vols. I and II (D. N. Glover, ed., 1985), IRL Press, Oxford, whole of text; Oligonucleotide Synthesis: A Practical Approach (M. J. Gait, ed, 1984) IRL Press, Oxford, whole of text, and particularly the papers therein by Gait, pp1-5, & 7-46; Atkinson et al, pp35-81; Sproat et al, pp 83-115; and Wu et al, pp 135-151; 4. Nucleic Acid Hybridization: A Practical Approach (B. D. Hames & S. J. Higgins, eds., 1985) IRL Press, Oxford, whole of text; Immobilized Cells and Enzymes: A Practical Approach (1986) IRL Press, Oxford, whole of text; Perbal, B., A Practical Guide to Molecular Cloning (1984); Methods In Enzymology (S. Colowick and N. Kaplan, eds., Academic Press, Inc.), whole of series; J. F. Ramalho Ortigao, “The Chemistry of Peptide Synthesis” In: Knowledge database of Access to Virtual Laboratory277702-566574 website (Interactiva, Germany); Sakakibara, D., Teichman, J., Lien, E. Land Fenichel, R. L. (1976). Biochem. Biophys. Res. Commun.73336-342; Merrifield, R. B. (1963). J. Am. Chem. Soc.85, 2149-2154; Barany, G. and Merrifield, R. B. (1979) in The Peptides (Gross, E. and Meienhofer, 3. eds.), vol.2, pp.1-284, Academic Press, New York.12. Wiinsch, E., ed. (1974) Synthese von Peptiden in Houben-Weyls Metoden der Organischen Chemie (Muler, E., ed.), vol.15, 4th edn., Parts 1 and 2, Thieme, Stuttgart; Bodanszky, M. (1984) Principles of Peptide Synthesis, Springer-Verlag, Heidelberg; Bodanszky, M. & Bodanszky, A. (1984) The Practice of Peptide Synthesis, Springer-Verlag, Heidelberg; Bodanszky, M. (1985) Int. J. Peptide Protein Res.25, 449-474; Handbook of Experimental Immunology, Vols. I-IV (D. M. Weir and C. C. Blackwell, eds., 1986, Blackwell Scientific Publications); and Animal Cell Culture: Practical Approach, Third Edition (John R. W. Masters, ed., 2000); each of these references are incorporated herein by reference in their entireties.

[0272] Although the disclosure of the invention has been described in detail forpurposes of clarity and understanding, it will be obvious to those with skill in the art that certain modifications can be practiced within the scope of the appended claims. All publications and patent documents cited herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each were so individually denoted.

[0273] Throughout this specification, unless the context requires otherwise, the word“comprise,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated step or element or integer or group of steps or elements or integers but not the exclusion of any other step or element or integer or group of elements or integers.

[0274] All patent applications, patents, and printed publications referred to hereinare incorporated by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety. And, all patent applications, patents, and printed publications cited herein are incorporated herein by reference in the entireties, except for any definitions, subject matter disclaimers, or disavowals, and except to the extent that the incorporated material is inconsistent with the express disclosure herein, in which case the language in this disclosure controls.

[0275] WILD-TYPE U1-AGATOXINS AND TVPS

[0276] “Hobo spiders” (Eratigena agrestis, formerly Tegenaria agrestis) arevenomous spiders that are members of the Agelenidae family of spiders, or funnel web weavers. See Ingale A, Antigenic epitopes prediction and MHC binder of a paralytic insecticidal toxin (ITX-1) of Tegenaria agrestis (hobo spider).4 August 2010 Volume277702-566574 2010:2 pp 97-103. The venom of Hobo spiders has been implicated as possessing insecticidal activity. See Johnson et al., Novel insecticidal peptides from Tegenaria agrestis spider venom may have a direct effect on the insect central nervous system. Arch Insect Biochem Physiol. 1998;38(1):19-31; Klint et al., Production of Recombinant Disulfide-Rich Venom Peptides for Structural and Functional Analysis via Expression in the Periplasm of E. coli. PLoS One. 2013; 8(5): e63865.

[0277] The Hobo spider—along with several other spiders in the Agelenidae family,produce venom containing agatoxins—which exhibit insecticidal activity. Agatoxins are a chemically diverse group of toxins that can induce various insecticidal effects depending on the target species; e.g., agatoxins cause slow-onset spastic paralysis in coleopterans, lepidopterans, and dipterans; increase the rate of neuron firing in the central nervous system (CNS) of houseflies (Musca domestica); and are lethal to other insects (e.g., the blowfly, Lucilia cuprina). Accordingly, agatoxins are implicated in targeting the CNS. See Undheim et al., Weaponization of a hormone: convergent recruitment of hyperglycemic hormone into the venom of arthropod predators. Structure 23: 1283-1292, and Johnson et al., Novel insecticidal peptides from Tegenaria agrestis spider venom may have a direct effect on the insect central nervous system. Arch. Insect Biochem. Physiol.38:19-31(1998).

[0278] Two types of agatoxins include U1-agatoxin-Ta1a and U1-agatoxin-Ta1b,which are both members of the helical arthropod-neuropeptide-derived (HAND) toxins family. In addition to spiders, these toxins can also be found in the venom of centipedes. The agatoxins are evolutionary offshoots of an ancient ecdysozoan hormone family, i.e., the ion transport peptide / crustacean hyperglycemic hormone (ITP / CHH) family. See Undheim et al., Weaponization of a hormone: convergent recruitment of hyperglycemic hormone into the venom of arthropod predators. Structure 23: 1283-1292, and Johnson et al., Novel insecticidal peptides from Tegenaria agrestis spider venom may have a direct effect on the insect central nervous system. Arch. Insect Biochem. Physiol.38:19-31(1998).

[0279] The Hobo-spider-derived U1-agatoxin-Ta1b toxin has a full amino acidsequence of “MKLQLMICLVLLPCFFCEPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRN DVYTACHEAQKG” (SEQ ID NO: 2)” which includes a signal peptide from amino acid positions 1-17, and the mature toxin from positions 18-68. Id. The protein comprises four tightly packed α-helices, with no β-strands present, and the molecular mass of the mature toxin is 5700.39 Daltons (Da). Id.277702-566574

[0280] An exemplary mature wild-type U1-agatoxin-Ta1b polypeptide fromEratigena agrestis is provided having the amino acid sequence: “EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG” (SEQ ID NO:1).

[0281] During protein processing, the mature wild-type U1-agatoxin-Ta1b toxinundergoes an excision event of the C-terminal glycine, yielding the following amino acid sequence: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3). A subsequent post-translational event result in the mature wild-type U1-agatoxin- Ta1b toxin having a C-terminal amidation.

[0282] The polypeptide as defined by its amino acid sequence as shown in SEQ IDNO: 4 is a proteolytically stable U1-Agatoxin-Ta1b variant polypeptide exemplified and described as SEQ ID NO: 51 in Applicant's PCT International Application PCT / US2021 / 028254, the disclosure of which is incorporated herein in its entirety. SEQ ID NO: 4 has an amino acid sequence of: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG. The SEQ. ID. NO: 4 polypeptide can serve as a control when comparing to the TVPs of the present disclosure in the Example experiments shown infra.

[0283] U1-agatoxin-Ta1b Variant Polypeptides (TVPs) are mutants or variants thatdiffer from the wild-type U1-agatoxin-Ta1b (SEQ ID NO:1) and Ta1b mutant polypeptide (SEQ ID NO: 4) in some way, e.g., in some embodiments, this variance can be an amino acid substitution, deletion, or addition; or a change to the polynucleotide encoding the wild-type U1-agatoxin-Ta1b resulting in an amino acid substitution, deletion, or addition. The result of this variation is a non-naturally occurring polypeptide and / or polynucleotide sequence encoding the same that possesses enhanced insecticidal activity against one or more insect species relative to the wild-type U1-agatoxin-Ta1b.

[0284] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100%277702-566574 identical to a compound or an agriculturally acceptable salt thereof of Formula I: (all of the variable letters Xnand Bnand Znare each independently amino acids in single letter format) X1-X2-X3-X4-X5-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A- E-C-F-R-N-D-V-Y-B2-A-C-Z1-Z2-Z3-Z4-Z5-Z6(SEQ ID NO: 98); wherein X1is E, or absent, X2 is P or absent, X3 is D or absent, X4 is E, or absent, X5 is I or absent, B1 is R or Q, B2 is T or A; Z1can be H or absent; Z2is E or absent; Z3is A or absent; Z4is Q or absent; Z5is K or absent; and Z6 is G or absent, and wherein the compound of Formula I does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3) or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10).

[0285] In some embodiments, present disclosure provides for U1-agatoxin-Ta1bvariant polypeptide (TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula II or an agriculturally acceptable salt thereof, (all of the variable letters Xnand Bnare each independently amino acids in single letter format) X1-X2-X3-X4-X5-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A- E-C-F-R-N-D-V-Y-B2-A-C-H-E-A-Q-K-G (SEQ ID NO: 99); wherein X1 is E, or absent, X2 is P or absent, X3is D or absent, X4is E, or absent, X5is I or absent, B1is R or Q, and B2is T or A; and wherein the compound of Formula II does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3) or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ277702-566574 ID NO: 1); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10).

[0286] In some embodiments, present disclosure provides for U1-agatoxin-Ta1bvariant polypeptide (TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula III, or an agriculturally acceptable salt thereof: (all of the variable letters Zn and Bn are each independently amino acids in single letter format) E-P-D- E-I-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A-E-C-F-R-N-D- V-Y-B2-A-C-Z1-Z2-Z3-Z4-Z5-Z6 (SEQ ID NO: 100); wherein B1 is R or Q, and B2 is T or A; Z1 is H or absent; Z2 is E or absent; Z3 is A or absent; Z4 is Q or absent; Z5 is K or absent; andZ6 is G or absent; and wherein the compound of Formula III does not consist of the aminoacid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3) or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10).

[0287] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure comprises, consists essentially of, or consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of:277702-566574 Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21);277702-566574 Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37);277702-566574 Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); or an agriculturally acceptable salt thereof, wherein the TVPs described herein have been shown to have a knockdown of 50% of the population concentration (KD50) of lower than 182 pmol / g against houseflies at 4 hours.

[0288] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or277702-566574 more insect species. TVPs of the present disclosure comprises, consists essentially of, or consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19);277702-566574 Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35);277702-566574 Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions. The TVPs described herein have been shown to have a knockdown of 50% of the population concentration (KD50) of lower than 182 pmol / g against houseflies at 24-hours.277702-566574

[0289] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 1: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 5.

[0290] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 2: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 6.

[0291] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 3: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 7.277702-566574

[0292] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 4: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 8.

[0293] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 9); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 9.

[0294] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 7: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 11.277702-566574

[0295] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12);wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 12.

[0296] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 9: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 13.

[0297] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 14.277702-566574

[0298] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 15.

[0299] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 12: PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 16.

[0300] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 13: DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 17.277702-566574

[0301] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 14: EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 18.

[0302] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 15: ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 19.

[0303] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 16: CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 20.277702-566574

[0304] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 17: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 21.

[0305] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 18: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 22.

[0306] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 19: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 23.277702-566574

[0307] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 20: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 24.

[0308] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 21: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 25.

[0309] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 22: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 26.277702-566574

[0310] In addition, the present disclosure describes a composition comprising a TVP,and one or more excipients; wherein the TVP comprises, consists essentially of, or consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19);277702-566574 Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35);277702-566574 Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5 and 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; and wherein the one or more excipients is selected from the group consisting of: trehalose; maltodextrin; maltose; potassium phosphate dibasic (K2HPO4); potassium phosphate monobasic (KH2PO4);277702-566574 lignosulfonate; gypsum; sorbitol; sodium benzoate; potassium sorbate; EDTA; benzisothiazolinone (BIT); and fermentation solids.

[0311] In addition, present disclosure describes a composition comprising a TVP, andone or more excipients; wherein the TVP consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18);277702-566574 Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34);277702-566574 Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; wherein the277702-566574 composition comprises an amount of TVP that is 8.5% wt / wt of the total weight of the composition; and wherein the plurality of excipients consists of the following: an amount of trehalose that is 25% wt / wt; an amount of BIT that is 0.05% wt / wt; an amount of maltodextrin that is 36.3% wt / wt; an amount of potassium phosphate dibasic anhydrous (K2HPO4) that is 2.6% wt / wt; an amount of potassium phosphate monobasic (KH2PO4) that is 0.4% wt / wt; and an amount of fermentation solids that is 26.85% wt / wt, of the total weight of the composition.

[0312] In addition, the present disclosure describes a polynucleotide encoding a TVP,the TVP comprises, consists essentially of, or consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16);277702-566574 Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32);277702-566574 Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48);277702-566574 Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5 and 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; or a complementary nucleotide sequence thereof.

[0313] In addition, the present disclosure describes a method of producing a TVP, themethod comprising: (a) preparing a vector comprising a first expression cassette comprising a polynucleotide operable to encode a TVP, or complementary nucleotide sequence thereof, said TVP comprises an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15);277702-566574 Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31);277702-566574 Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47);277702-566574 Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; (b) introducing the vector into a host cell; and (c) growing the host cell in a growth medium under conditions operable to enable expression of the TVP and secretion into the growth medium.

[0314] In addition, the present disclosure describes a method of using thecompositions described above to control insects comprising, providing the compositions to the locus of an insect.

[0315] In addition, the present disclosure describes a method of protecting a plantfrom insects comprising, providing a plant which expresses a TVP, or polynucleotide encoding the same.

[0316] In addition, the present disclosure describes a method of combating,controlling, or inhibiting a pest comprising, applying a pesticidally effective amount of the compositions described above to the locus of the pest, or to a plant or animal susceptible to an attack by the pest.

[0317] In addition, the present disclosure describes a vector comprising apolynucleotide operable to encode a TVP comprising, consisting essentially of, or consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9);277702-566574 Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26);277702-566574 Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42);277702-566574 Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5 and 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions.

[0318] In addition, the present disclosure describes a yeast strain comprising: a firstexpression cassette comprising a polynucleotide operable to encode a TVP, said TVP consisting of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9);277702-566574 Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26);277702-566574 Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42);277702-566574 Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5 and 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions; or complementary nucleotide sequence thereof.

[0319] In some embodiments, an insecticidal U1-agatoxin-Ta1b variant polypeptide(TVP) consists of an amino acid sequence of SEQ ID NO: 5.

[0320] In some embodiments, an insecticidal U1-agatoxin-Ta1b variant polypeptide(TVP) consists of an amino acid sequence of SEQ ID NO: 6.

[0321] In some embodiments, an insecticidal U1-agatoxin-Ta1b variant polypeptide(TVP) consists of an amino acid sequence of SEQ ID NO: 7.

[0322] In some embodiments, an insecticidal U1-agatoxin-Ta1b variant polypeptide(TVP) consists of an amino acid sequence of SEQ ID NO: 8.

[0323] In some embodiments, an insecticidal U1-agatoxin-Ta1b variant polypeptide(TVP) consists of an amino acid sequence of SEQ ID NO: 9.

[0324] In some embodiments, a TVP comprises, consists essentially of, or consists ofan amino acid sequence according to SEQ ID NOs: 5-9 and 11-50, as shown in Table 1.277702-566574 editO oeNuescT C A Y E Q K A K S E G N C V F T C A o F A E T C A Y E Q K A K S E G N C F A lneE c u E A H F A E T C A Y E Q K A K S E G E A H s K V C E A H F A E T C A Y E Q K A K K V C iqN Q A K V C E A H F A E T deC A Y E Q N Q A T D T N Q A K V C E A H F A E T C A T tSD A n d M G Y T D A N Q A K V C E A H F A E M G Y R C V M G esicY T D A N Q A K V C E A H Q C V A N D Q C V M G Y T D A N Q A K V C A N D e A R N N A N D Q C V M G Y T D A N Q A R N N roC C R R N N A N D Q C V M G Y T D A C C R p eniI V F C C R R N N A N D Q C V M G Y I V F E N C I V F C C R R N N A N D Q C V E N C h m D S E G E N C I V F C C R R N N A N D D S E t A P K A K D S E G E N C I V F C C R R N N P K A f E Y E Q P K A K D S E G E N C I V F C C R E Y E Q osP Ve 1d 2 3 4 5 7Tbnd d d d d. unun n n n1 ma1aelNT-ou u u upopopopopopbaTm m m m m mT WoCoCoCoCoCoC dicO AN]5 oDI1 5 6 7 8 9112ni3 Q 0 mE 0[A S277702-566574ecT C A T C T C T C Y E Q K A K S E G N C neF A E F A E F A F A T C A Y E Q K A K S E G u E A H E A H E A H E A F A E T C A Y E Q K A K q K V C K V C K V C K V C E A H F A E T C A Y E Q e N Q A N Q A N Q A N Q A K V C E A H F A E T C A S T D A T D A T D A T D A N Q A K V C E A H F A E d M G Y M G Y M G Y M G Y T D T N Q A K V C E A H icQ C V Q C V Q C V Q C V M G Y T D T N Q A K V C A N D A N D A N D A N D R C V M G Y T D T N Q A A R N N R N N R N N R N N A N D R C V M G Y T D T o C C R C C R C C R C C R R N N A N D R C V M G Y niI V F I V F I V F I V F C C R R N N A N D R C V E N C E N C E N C E N C I V F C C R R N N A N D m D S E D S E D S E D S E E N C I V F C C R R N N A P K A P K A P K A P K A D S E G E N C I V F C C R E Y E E Y E E Y E E Y E P K A K D S E G E N C I V F 01 2 3 4 5e 8d 91 1 1 1 1 1 mandundndndndndndnouououououououNp p p p p p popmomomomomomom mC C C C C CoCoC dicO ANoDniI 2131415161718191mQ AE S277702-566574ecV F T C A T C A T C A T C T C T C Y E Q neN C F A E F A E F A E F A E F A F A T C A u S E G E A H E A H E A H E A H E A H E A F A E q K A K K V C K V C K V C K V C K V C K V C E A H e Y E Q N Q A N Q A N Q A N Q A N Q A N Q A K V C S T C A T D T T D T T D T T D T T D T T D T N Q A d F A E M G Y M G Y M G Y M G Y M G Y M G Y T D A icE A H R C V R C V R C V R C V R C V R C V M G Y K V C A N D A N D A N D A N D A N D A N D Q C V A N Q A R N N R N N R N N R N N R N N R N N A N D o T D T C C R C C R C C R C C R C C R C C R R N N niM G Y I V F I V F I V F I V F I V F I V F C C R R C V E N C E N C E N C E N C E N C E N C I V F m A N D D S E D S E D S E D S E D S E D S E E N C A R N N P K A K P K A P K A P K A P K A P K A D S E C C R E Y E Q E Y E Q E Y E E Y E E Y E E Y E P K A K 67 8 9 0 1 2 3e1 d 1 1 1 2 2 2 2 mandundndndndndndnouououououououNp p p p p p popmomomomomomom mC C C C C CoCoC dicO ANoDniI 0212223242526272mQ AE S277702-566574ecK A K S E N C Y E Q K A S E N C Y E neY E Q K A K S E T C A Y E Q K A S E T C A u T C A Y E Q K A K F A E T C A Y E Q K A F A E q F A E T C A Y E Q E A H F A E T C A Y E Q E A H e E A H F A E T C A K V C E A H F A E T C A K V C S K V C E A H F A E N Q A K V C E A H F A E N Q A d N Q A K V C E A H T D A N Q A K V C E A H T D A icT D A N Q A K V C M G Y T D A N Q A K V C M G Y M G Y T D A N Q A Q C V M G Y T D A N Q A Q C V A Q C V M G Y T D A A N D Q C V M G Y T D A A N D o A N D Q C V M G Y R N N A N D Q C V M G Y R N N niR N N A N D Q C V C C R R N N A N D Q C V C C R C C R R N N A N D I V F C C R R N N A N D I V F m I V F C C R R N N E N C I V F C C R R N N E N C A E N C I V F C C R D S E E N C I V F C C R D S E D S E E N C I V F P K A D S E E N C I V F P K A 425 6 7 8 9 0 1e d2 2 2 2 2 3 3 mandundndndndndndnouououououououNp p p p p p popmomomomomomom mC C C C C CoCoC dicO ANoDniI 8292031323334353mQ AE S277702-566574neY E K A S E T C Y E K A N C T C Y E K A u T C A Y E K A F A E T C Y E S E F A T C Y E q F A E T C A Y E E A H F A E T C K A E A H F A T C e E A H F A E T C A K V C E A H F A E Y E K V C E A H F A S K V C E A H F A E N Q A K V C E A H T C N Q A K V C E A H d N Q A K V C E A H T D A N Q A K V C F A E T D A N Q A K V C icT D A N Q A K V C M G Y T D A N Q A E A H M G Y T D A N Q A M G Y T D A N Q A Q C V M G Y T D A K V C Q C V M G Y T D A A Q C V M G Y T D A A N D Q C V M G Y N Q A A N D Q C V M G Y o A N D Q C V M G Y R N N A N D Q C V T D A R N N A N D Q C V niR N N A N D Q C V C C R R N N A N D M G Y C C R R N N A N D C C R R N N A N D I V F C C R R N N Q C V I V F C C R R N N m I V F C C R R N N E N C I V F C C R A N D E N C I V F C C R A E N C I V F C C R D S E E N C I V F R N N D S E E N C I V F D S E E N C I V F P K A D S E E N C C C R P K A D S E E N C 23334353637383930 1e d d d d d d d4 4 man n n n n nd d duNouou u u ununununup popopopopopopopopmomomomomomomomom mC C C C C C C CoCoC dicO ANoDniI 63738393041424344454mQ AE S277702-566574eG G G A G G A G T A C g G G G u q A G a G T C C C T C C C A G a e A A c T G A C G T A A A A A c S A C g A T G A G G A G T A C g e C C C a T T G T C C C T C C C d A A A a A A T G A C G T A A A itA G T c A A A T G A G G A G T oeC C T A T T a T T G T C C C T l C G T C A G a A A T G A C G T c A G G G A C c A A A T G A G G u G T C A C T A T T a T T G T C N T G A G G T C A G a A A T G A A T G A T T G A C c A A A T G a T T C A C A C T A T T a T T a A A G T G G G T C A G a A A c A A T G T A T T G A C c A A A T T A C G C A C A C T A T T C A G T A A G T G G G T C A G G A C A A G T G T A T T G A C A C T G C A A C G C A C A C T G G T A C C T A A G T G G G T A T T G T G A A G T G T A T T C A C C G G G C A A C G C A C G T G A A A A C C T A A G T G T G T G A G G T G A A G T G T A C G A T C C G G G C A A C G T A A C A G A A A A C C T A A A A G C T T G A G G T G A A G ecN C Y E K A S E N C neS E T C Y E K A S E u K A F A T C Y E K A q Y E E A F A T C Y E e T C K V C E A F A T C S F A N Q A K V C E A F A d E A H T D A N Q A K V C E A icK V C M G Y T D A N Q A K V C N Q A Q C V M G Y T D A N Q A A T D A A N D Q C V M G Y T D A o M G Y R N N A N D Q C V M G Y niQ C V C C R R N N A N D Q C V A N D I V F C C R R N N A N D m R N N E N C I V F C C R R N N A C C R D S E E N C I V F C C R I V F P K A D S E E N C I V F 4:34445 6e d d d4 4 nn ndndma uououounouNp p p popmom m m mCoCoCoCoC dicO ANoDniI 6474849405mQ AE S277702-566574

[0326] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure have an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of Table 1 along the entire length of the TVP.

[0327] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure consist of an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of Table 1 along the entire length of the TVP.

[0328] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. TVPs of the present disclosure comprise, consist essentially of, or consists of an amino acid sequence along its entire length, that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7);277702-566574 Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24);277702-566574 Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40);277702-566574 Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5 and 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions.

[0329] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 1: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 5.277702-566574

[0330] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 2: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 6.

[0331] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 3: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 7.

[0332] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 4: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 8.277702-566574

[0333] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 9); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 9.

[0334] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 7: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 11.

[0335] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 8: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 12.277702-566574

[0336] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 9: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 13.

[0337] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 14.

[0338] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 15.277702-566574

[0339] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 12: PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 16.

[0340] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 13: DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 17.

[0341] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 14 EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 18.277702-566574

[0342] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 15: ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 19.

[0343] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 16: CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 20.

[0344] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 17: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 21.277702-566574

[0345] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 18: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 22.

[0346] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 19: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 23.

[0347] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 20: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 24.277702-566574

[0348] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 21: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 25.

[0349] The present disclosure provides for U1-agatoxin-Ta1b variant polypeptide(TVP), compositions comprising a TVP, insecticidal proteins comprising one or more TVPs optionally with other proteins, and methods for their use to eradicate, kill, control, inhibit, injure, confuse, render sterile, or combinations thereof, one or more pests. The TVPs described herein have pesticidal activity, for example, insecticidal activity against one or more insect species. In some embodiments, a TVP of the present disclosure has an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to the amino acid sequence of: Compound 22: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); wherein the amino acid sequence of the TVP has 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions relative to SEQ ID NO: 26.

[0350] In various embodiments, polynucleotides encoding TVPs can be used totransform plant cells, yeast cells, or bacteria cells. In some embodiments, the insecticidal TVP transgenic proteins may be formulated into compositions that can be sprayed or otherwise applied in any manner known to those skilled in the art to the surface of plants or parts thereof. Accordingly, DNA constructs are provided herein, operable to encode one or more TVPs under the appropriate conditions in a host cell, for example, a plant cell. Methods for controlling a pest infection by a parasitic insect of a plant cell comprises administering or introducing a polynucleotide encoding a TVP as described herein to a plant, plant tissue, or a plant cell by recombinant techniques and growing said recombinantly altered plant, plant tissue or plant cell in a field exposed to the pest. Alternatively, TVPs can be formulated into a sprayable composition consisting of a TVP and an excipient, and applied directly to277702-566574 susceptible plants by direct application, such that upon ingestion of the TVP by the infectious insect results in a deleterious effect.

[0351] In some embodiments, a TVP comprises, consists essentially of, or consists of,an amino sequence that is at least 50% identical, at least 55% identical, at least 60% identical, at least 65% identical, at least 70% identical, at least 75% identical, at least 80% identical, at least 81% identical, at least 82% identical, at least 83% identical, at least 84% identical, at least 85% identical, at least 86% identical, at least 87% identical, at least 88% identical, at least 89% identical, at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, at least 99.5% identical, at least 99.6% identical, at least 99.7% identical, at least 99.8% identical, at least 99.9% identical, or 100% identical to an amino acid sequence as set forth in any one of SEQ ID NOs: 5-9 and 11-50, or an agriculturally acceptable salt thereof; with the proviso that the TVP does not consist of the amino acid sequence of SEQ ID NOs: 1, 3, or 4.

[0352] In some embodiments, the TVP may comprise an amino acid sequence that isat least 50% identical, at least 55% identical, at least 60% identical, at least 65% identical, at least 70% identical, at least 75% identical, at least 80% identical, at least 81% identical, at least 82% identical, at least 83% identical, at least 84% identical, at least 85% identical, at least 86% identical, at least 87% identical, at least 88% identical, at least 89% identical, at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, at least 99.5% identical, at least 99.6% identical, at least 99.7% identical, at least 99.8% identical, at least 99.9% identical, or 100% identical to an amino acid sequence set forth in SEQ ID NOs: 5-9 and 11-50.

[0353] In some embodiments, the TVP may comprise an amino acid sequence that isat least 50% identical, at least 55% identical, at least 60% identical, at least 65% identical, at least 70% identical, at least 75% identical, at least 80% identical, at least 81% identical, at least 82% identical, at least 83% identical, at least 84% identical, at least 85% identical, at least 86% identical, at least 87% identical, at least 88% identical, at least 89% identical, at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, at least 99.5% identical, at least 99.6% identical, at least 99.7% identical, at least 99.8% identical, at least 99.9% identical, or 100% identical to277702-566574 an amino acid sequence: Compound 2: “DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG” (SEQ ID NO: 6).

[0354] In some embodiments, the TVP may comprise an amino acid sequence that isat least 50% identical, at least 55% identical, at least 60% identical, at least 65% identical, at least 70% identical, at least 75% identical, at least 80% identical, at least 81% identical, at least 82% identical, at least 83% identical, at least 84% identical, at least 85% identical, at least 86% identical, at least 87% identical, at least 88% identical, at least 89% identical, at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, at least 99.5% identical, at least 99.6% identical, at least 99.7% identical, at least 99.8% identical, at least 99.9% identical, or 100% identical to an amino acid sequence: Compound 3: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7).

[0355] In some embodiments, a TVP can be encoded by a polynucleotide. Forexample, in some embodiments, a polynucleotide encoding a TVP of the present disclosure wherein the TVP can comprise an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to at least one of the amino acid sequences of: Compound 1:PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2:DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3:EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4:ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8);Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ IDNO: 9); Compound 7:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12);277702-566574 Compound 9:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12 PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13:DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14:EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15:ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16:CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17:EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22:EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28);277702-566574 Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44);277702-566574 Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); wherein each of the amino acid sequences of Compounds 1-5, & 7-46 each independently have 0, 1, 2, 3, 4, or 5 conservative amino acid substitutions.; or a complementary nucleotide sequence thereof.

[0356] In some embodiments, a polynucleotide encoding a TVP can comprise apolynucleotide, wherein the TVP comprises an amino sequence as set forth in any one of SEQ ID NOs: 51-55 and 57-96.

[0357] In some embodiments, a polynucleotide encoding a TVP can comprise apolynucleotide, wherein the polynucleotide sequence has a sequence as set forth in any one of SEQ ID NOs: 51-55 and 57-96, or a complementary nucleotide sequence thereof.

[0358] In various embodiments, polynucleotides encoding TVPs can be used totransform plant cells, yeast cells, or bacteria cells. In some embodiments, the insecticidal TVP transgenic proteins may be formulated into compositions that can be sprayed or otherwise applied in any manner known to those skilled in the art to the surface of plants or parts thereof. Accordingly, DNA constructs are provided herein, operable to encode one or more TVPs under the appropriate conditions in a host cell, for example, a plant cell. Methods for controlling a pest infection by a parasitic insect of a plant cell comprises administering or introducing a polynucleotide encoding a TVP as described herein to a plant, plant tissue, or a plant cell by recombinant techniques and growing said recombinantly altered plant, plant tissue or plant cell in a field exposed to the pest. Alternatively, TVPs can be formulated into a sprayable composition consisting of a TVP and an excipient, and applied directly to susceptible plants by direct application, such that upon ingestion of the TVP by the infectious insect results in a deleterious effect.277702-566574

[0359] In some embodiments, a TVP may have an amino acid sequence of any one ofSEQ ID NOs: 5-9 and 11-50. In some embodiments, the TVP may comprise, consist essentially of, or consist of an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% amino acid sequence identity to of SEQ ID NOs: 5-9 and 11-50.

[0360] In some embodiments, the TVP may comprise, consist essentially of, orconsist of an amino acid sequence that is at least 50% identical, at least 55% identical, at least 60% identical, at least 65% identical, at least 70% identical, at least 75% identical, at least 80% identical, at least 81% identical, at least 82% identical, at least 83% identical, at least 84% identical, at least 85% identical, at least 86% identical, at least 87% identical, at least 88% identical, at least 89% identical, at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, at least 99.5% identical, at least 99.6% identical, at least 99.7% identical, at least 99.8% identical, at least 99.9% identical, or 100% identical to an amino acid sequence set forth in SEQ ID NOs: 5-9 and 11-50.

[0361] In some embodiments, the TVP may comprise, consist essentially of, orconsist of an amino acid sequence that is at least 50% identical, at least 55% identical, at least 60% identical, at least 65% identical, at least 70% identical, at least 75% identical, at least 80% identical, at least 81% identical, at least 82% identical, at least 83% identical, at least 84% identical, at least 85% identical, at least 86% identical, at least 87% identical, at least 88% identical, at least 89% identical, at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, at least 99.5% identical, at least 99.6% identical, at least 99.7% identical, at least 99.8% identical, at least 99.9% identical, or 100% identical to an amino acid sequence: Compound 2: “DEICRAQMTNKE...

Claims

277702-566574 CLAIMS 1. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP comprising an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula I: (all of the variable letters Xn and Bnand Znare each independently amino acids in single letter format) X1-X2-X3-X4-X5-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A- E-C-F-R-N-D-V-Y-B2-A-C-Z1-Z2-Z3-Z4-Z5-Z6(SEQ ID NO: 98); wherein X1is E, or absent, X2 is P or absent, X3 is D or absent, X4 is E, or absent, X5 is I or absent, B1 is R or Q, B2 is T or A; Z1can be H or absent; Z2is E or absent; Z3is A or absent; Z4is Q or absent; Z5is K or absent; and Z6 is G or absent, and wherein the compound of Formula I does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10); or an agriculturally acceptable salt thereof.

2. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP comprising an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula II: (all of the variable letters Xn and Bnare each independently amino acids in single letter format) X1-X2-X3-X4-X5-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A- E-C-F-R-N-D-V-Y-B2-A-C-H-E-A-Q-K-G (SEQ ID NO: 99); wherein X1 is E, or absent, X2 is P or absent, X3is D or absent, X4is E, or absent, X5is I or absent, B1is R or Q, and B2is T or A; and wherein the compound of Formula II does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID277702-566574 NO: 3); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10); or an agriculturally acceptable salt thereof.

3. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP comprising an amino acid sequence that is at least 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound of Formula III, or an agriculturally acceptable salt thereof: (all of the variable letters Zn and Bn are each independently amino acids in single letter format) E-P-D-E-I-C-R-A-B1-M-T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C- G-D-Q-V-A-A-C-E-A-E-C-F-R-N-D-V-Y-B2-A-C-Z1-Z2-Z3-Z4-Z5-Z6(SEQ ID NO: 100); wherein B1 is R or Q, and B2 is T or A; Z1 is H or absent; Z2 is E or absent; Z3 is A or absent; Z4 is Q or absent; Z5 is K or absent; and Z6 is G or absent; and wherein the compound of Formula III does not consist of the amino acid sequence: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 4); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 1); or EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 3); or EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 10); or an agriculturally acceptable salt thereof.

4. The TVP of any one of claims 1-3, wherein the TVP consists of an amino sequence as set forth in a compound selected from: Compound 1: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8); Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 9); Compound 7: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12: PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16);277702-566574 Compound 13: DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14: EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15: ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16: CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41); Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50) 5. The TVP of claim 4, wherein the TVP is encoded by a polynucleotide sequence as set forth in any one of SEQ ID NOs: 51-55, and 57-96, or a complementary nucleotide sequence thereof.277702-566574 6. The TVP of any one of claims 1-4, wherein the TVP further comprises a homopolymer or heteropolymer of two or more TVPs, wherein the amino acid sequence of each TVP is the same or different.

7. The TVP of any one of claims 1-4, wherein the TVP is a fused protein comprising two or more TVPs separated by a cleavable linker or a non-cleavable linker, and wherein the amino acid sequence of each TVP may be the same or different.

8. The TVP of claim 7, wherein the linker is a cleavable linker.

9. The TVP or claim 7, wherein the cleavable linker is cleavable inside the gut or hemolymph of an insect.

10. The TVP of claim 9, wherein the linker has an amino acid sequence as set forth in any one of SEQ ID NOs: 101-104 and 112-120.

11. A composition or formulation comprising a TVP of any one of claims 1-4, or an agriculturally acceptable salt thereof, and one or more excipients; wherein the one or more excipients is selected from the group consisting of: trehalose; maltodextrin; maltose; potassium phosphate dibasic (K2HPO4); potassium phosphate monobasic (KH2PO4); lignosulfonate; gypsum; sorbitol; sodium benzoate; potassium sorbate; EDTA; benzisothiazolinone (BIT); and fermentation solids.

12. The composition or formulation of claim 11, wherein the one or more excipients is selected from the group consisting of: trehalose; maltodextrin; potassium phosphate dibasic anhydrous (K2HPO4); potassium phosphate monobasic (KH2PO4); BIT; and fermentation solids.

13. The composition or formulation of claim 12, wherein the TVP ranges from about 2% to about 16% w / w; and wherein trehalose ranges from about 5% to about 40% w / w; BIT ranges from about 0.01% to about 0.1% w / w; maltodextrin ranges from about 10% to about 50% w / w; potassium phosphate dibasic anhydrous (K2HPO4) ranges from about 1% to about 5% w / w; and potassium phosphate monobasic (KH2PO4) ranges from about 0.10% to about277702-566574 1% w / w; and fermentation solids range from about 15% to about 40% w / w; of the total weight of the composition.

14. The composition or formulation of claim 13, wherein the TVP ranges from about 7% to about 9% w / w; and wherein trehalose ranges from about 20% to about 30% w / w; BIT ranges from about 0.025% to about 0.075% w / w; maltodextrin ranges from about 30% to about 40% w / w; potassium phosphate dibasic anhydrous (K2HPO4) ranges from about 2% to about 3% w / w; potassium phosphate monobasic (KH2PO4) ranges from about 0.2% to about 0.6%; and fermentation solids range from about 20% to about 30%, of the total weight of the composition.

15. The composition or formulation of claim 14, wherein the TVP is about 8.5% w / w; and wherein trehalose is about 25% w / w; BIT is about 0.05% w / w; maltodextrin is about 36.3% w / w; potassium phosphate dibasic anhydrous (K2HPO4) is about 2.6% w / w; potassium phosphate monobasic (KH2PO4) is about 0.4% w / w; and fermentation solids are about 26.85% w / w, of the total weight of the composition.

16. The composition or formulation of claim 14, wherein the composition consists essentially of the following: an amount of TVP that is 8.5% w / w; an amount of trehalose that is 25% w / w; an amount of BIT that is 0.05% w / w; an amount of maltodextrin that is 36.3% w / w; an amount of potassium phosphate dibasic anhydrous (K2HPO4) that is 2.6% w / w; an amount of potassium phosphate monobasic (KH2PO4) that is 0.4% w / w; and an amount of fermentation solids that is 26.85% w / w, of the total weight of the composition.

17. The composition or formulation of claim 14, wherein the composition consists of the following: an amount of TVP that is 8.5% w / w; an amount of trehalose that is 25% w / w; an amount of BIT that is 0.05% w / w; an amount of maltodextrin that is 36.3% w / w; an amount of potassium phosphate dibasic anhydrous (K2HPO4) that is 2.6% w / w; an amount of potassium phosphate monobasic (KH2PO4) that is 0.4% w / w; and an amount of fermentation solids that is 26.85% w / w, of the total weight of the composition.

18. A composition or formulation consisting of a TVP, and a plurality of excipients; wherein the TVP consists of an amino acid sequence that is at least 80%, or 85%, or 90%, or277702-566574 95%, or 96%, or 97%, or 98%, or 99%, or at least 100% identical to a compound selected from: Compound 1: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 5); Compound 2: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 6); Compound 3: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 7); Compound 4: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 8); Compound 5: CRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQKG (SEQ ID NO: 9); Compound 7: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 11); Compound 8: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 12); Compound 9: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 13); Compound 10: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 14); Compound 11: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 15); Compound 12: PDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 16); Compound 13: DEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 17); Compound 14: EICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 18); Compound 15: ICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 19); Compound 16: CRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQKG (SEQ ID NO: 20); Compound 17: EPDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQK (SEQ ID NO: 21); Compound 18: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEAQ (SEQ ID NO: 22); Compound 19: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHEA (SEQ ID NO: 23); Compound 20: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACHE (SEQ ID NO: 24); Compound 21: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTACH (SEQ ID NO: 25); Compound 22: EPDEICRARMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYTAC (SEQ ID NO: 26); Compound 23: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 27); Compound 24: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 28); Compound 25: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 29); Compound 26: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQK (SEQ ID NO: 30); Compound 27: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 31); Compound 28: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 32); Compound 29: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 33); Compound 30: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEAQ (SEQ ID NO: 34); Compound 31: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 35); Compound 32: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 36); Compound 33: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 37); Compound 34: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHEA (SEQ ID NO: 38); Compound 35: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 39); Compound 36: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 40); Compound 37: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 41);277702-566574 Compound 38: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACHE (SEQ ID NO: 42); Compound 39: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 43); Compound 40: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 44); Compound 41: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 45); Compound 42: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAACH (SEQ ID NO: 46); Compound 43: PDEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 47); Compound 44: DEICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 48); Compound 45: EICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 49); and Compound 46: ICRAQMTNKEFTYKSNVCNNCGDQVAACEAECFRNDVYAAC (SEQ ID NO: 50); or an agriculturally acceptable salt thereof; wherein the composition or formulation consists of an amount of TVP that is 8.5% w / w of the total weight of the composition; and wherein the plurality of excipients consists of the following: an amount of trehalose that is 25% w / w; an amount of BIT that is 0.05% w / w; an amount of maltodextrin that is 36.3% w / w; an amount of potassium phosphate dibasic anhydrous (K2HPO4) that is 2.6% w / w; an amount of potassium phosphate monobasic (KH2PO4) that is 0.4% w / w; and an amount of fermentation solids that is 26.85% w / w, of the total weight of the composition.

19. A polynucleotide operable to encode a TVP of any one of claims 1-4; or a complementary nucleotide sequence thereof.

20. The polynucleotide of claim 19, wherein the polynucleotide sequence has a nucleotide sequence as set forth in any one of SEQ ID NOs: 51-55, and 57-96, or a complementary nucleotide sequence thereof.

21. A method of producing a TVP, the method comprising: (a) preparing a vector comprising a first expression cassette comprising a polynucleotide operable to encode a TVP of any one of claims 1-4; (b) introducing the vector into a host cell; and (c) growing the host cell in a growth medium under conditions operable to enable expression of the TVP and secretion into the growth medium.

22. The method of claim 21, wherein the vector is a plasmid comprising an alpha-MF signal.277702-566574 23. The method of claim 22, wherein the alpha-MF signal is operable to express an alpha- MF signal peptide.

24. The method of claim 21, wherein the vector is transformed into a host cell.

25. The method of claim 24, wherein the host cell is a eukaryotic cell or a prokaryotic cell.

26. The method of claim 25, wherein the eukaryotic cell is a yeast cell.

27. The method of claim 26, wherein the yeast cell is selected from any species belonging to the genera: Saccharomyces, Komagataella, Kluyveromyces, Pichia, Yarrowia, or Schizosaccharomyces.

28. The method of claim 27, wherein the yeast cell is selected from the group consisting of Kluyveromyces lactis, Kluyveromyces marxianus, Saccharomyces cerevisiae, and Komagataella phaffii.

29. The method of claim 27, wherein the yeast cell is Kluyveromyces lactis or Kluyveromyces marxianus.

30. The method of claim 28, wherein the yeast cell is Kluyveromyces lactis.

31. The method of claim 21, wherein the TVP is secreted into the growth medium.

32. The method of claim 31, wherein the TVP is operably linked to the alpha-MF signal peptide.

33. The method of any one of claims 21-26, wherein expression of the TVP in a yeast cell provides a yield of: at least 70 mg / L, at least 80 mg / L, at least 90 mg / L, at least 100 mg / L, at least 110 mg / L, at least 120 mg / L, at least 130 mg / L, at least 140 mg / L, at least 150 mg / L, at least 160 mg / L, at least 170 mg / L, at least 180 mg / L, at least 190 mg / L 200 mg / L, at least 500 mg / L, at least 750 mg / L, at least 1,000 mg / L, at least 1,250 mg / L, at least 1,500 mg / L, at least 1,750 mg / L, at least 2,000 mg / L, at least 2,500 mg / L, at least 3,000 mg / L, at least 3,500277702-566574 mg / L, at least 4,000 mg / L, at least 4,500 mg / L, at least 5,000 mg / L, at least 5,500 mg / L, at least at least 6,000 mg / L, at least 6,500 mg / L, at least 7,000 mg / L, at least 7,500 mg / L, at least 8,000 mg / L, at least 8,500 mg / L, at least 9,000 mg / L, at least 9,500 mg / L, at least 10,000 mg / L, at least 11,000 mg / L, at least 12,000 mg / L, at least 12,500 mg / L, at least 13,000 mg / L, at least 14,000 mg / L, at least 15,000 mg / L, at least 16,000 mg / L, at least 17,000 mg / L, at least 17,500 mg / L, at least 18,000 mg / L, at least 19,000 mg / L, at least 20,000 mg / L, at least 25,000 mg / L, at least 30,000 mg / L, at least 40,000 mg / L, at least 50,000 mg / L, at least 60,000 mg / L, at least 70,000 mg / L, at least 80,000 mg / L, at least 90,000 mg / L, or at least 100,000 mg / L of TVP per liter of yeast culture medium.

34. The method of claim 33, wherein expression of the TVP provides a yield of at least 100 mg / L of TVP per liter of medium.

35. The method of claim 33, wherein expression of the TVP in the medium results in the expression of a single TVP in the medium.

36. The method of claim 31, wherein expression of the TVP in the medium results in the expression of a TVP fusion polymer comprising two or more TVP polypeptides in the medium.

37. The method of claim 31, wherein the vector comprises two or three expression cassettes, each expression cassettes operable to encode the TVP of the first expression cassette.

38. The method of claim 31, wherein the vector comprises two or three expression cassettes, each expression cassettes operable to encode the TVP of the first expression cassette, or a TVP of a different expression cassettes.

39. The method of claim 31, wherein the first expression cassette is operable to encode a TVP as set forth in any one of SEQ ID NOs: 2-15, 49-53, or 77-110.

40. A method of using the composition or formulation of any one of claims 11-18 to control insects comprising: providing the composition or formulation of any one of claims 11-18 to the locus of an insect.277702-566574 41. The method of claim 40, wherein the insects are selected from the group consisting of: Achema Sphinx Moth (Hornworm) (Eumorpha achemon); Alfalfa Caterpillar (Coliaseurytheme); Almond Moth (Cadra cautella); Amorbia Moth (Amorbia humerosana);Armyworm (Spodoptera spp., e.g. exigua, frugiperda, littoralis, Mythimna unipuncta); Artichoke Plume Moth (Platyptilia carduidactyla); Azalea Caterpillar (Datana major);Bagworm (Thyridopteryx ephemeraeformis); Banana Moth (Hypercompe scribonia); BananaSkipper (Erionota thrax); Blackheaded Budworm (Acleris gloverana); California Oakworm (Phryganidia californica); Spring Cankerworm (Paleacrita merriccata); Cherry Fruitworm (Grapholita packardi); China Mark Moth (Nymphula nitidulata); Citrus Cutworm (Egira curialis); Codling Moth (Cydia pomonella); Cranberry Fruitworm (Acrobasis vaccinii); Cross-striped Cabbageworm (Evergestis rimosalis); Cutworm (Noctuid species, Agrotis ipsilon); Douglas Fir Tussock Moth (Orgyia pseudotsugata); Ello Moth (Hornworm) (Erinnyis ello); Elm Spanworm (Ennomos subsignaria); European Grapevine Moth (Lobesia botrana); European Skipper (Thymelicus lineola); Fall Webworm (Hyphantria cunea); Filbert Leafroller (Archips rosanus); Fruittree Leafroller (Archips argyrosplia); Grape Berry Moth (Paralobesia viteana); Grape Leafroller (Platynota stultana); Grapeleaf Skeletonizer (Harrisina americana); Green Cloverworm (Hypena scabra); Greenstriped Mapleworm (Dryocampa rubicunda); Gummosos (Batrachedra comosae); Gypsy Moth (Lymantria dispar); Hemlock Looper (Lambdina fiscellaria); Hornworm (Manduca spp.); Imported Cabbageworm (Pieris rapae); Io Moth (Automeris io); Jack Pine Budworm (Choristoneura pinus); Light Brown Apple Moth (Epiphyas postvittana); Melonworm (Diaphania hyalinata); Mimosa Webworm (Homadaula anisocentra); Obliquebanded Leafroller (Choristoneura rosaceana); Oleander Moth (Syntomeida epilais); Omnivorous Leafroller (Playnota stultana); Omnivorous Looper (Sabulodes aegrotata); Orangedog (Papilio cresphontes); Orange Tortrix (Argyrotaenia citrana); Oriental Fruit Moth (Grapholita molesta); Peach Twig Borer (Anarsia lineatella); Pine Butterfly (Neophasia menapia); Podworm (Helicoverpa zea); Redbanded Leafroller (Argyrotaenia velutinana); Redhumped Caterpillar (Schizura concinna); Rindworm Complex (Various Leps.); Saddleback Caterpillar (Acharia stimuli); Saddle Prominent Caterpillar (Heterocampa guttivitta); Saltmarsh Caterpillar (Estigmene acrea); Sod Webworm (Crambus spp.); Spanworm (Ennomos subsignaria ); Fall Cankerworm (Alsophila pometaria); Spruce Budworm (Choristoneura fumiferana); Tent Caterpillar (Various Lasiocampidae ); Fruit Borer Caterpillar (Thecla basilides); Tobacco Hornworm (Manduca sexta); Tobacco Moth (Ephestia elutella); Tufted Apple Budmoth277702-566574 (Platynota idaeusalis); Variegated Cutworm (Peridroma saucia); Variegated Leafroller (Platynota flavedana); Velvetbean Caterpillar (Anticarsia gemmatalis); Walnut Caterpillar (Datana integerrima); Western Tussock Moth (Orgyia vetusta); Southern Cornstalk Borer (Diatraea crambidoides); Corn Earworm (Helicoverpa armigera); Sweet Potato Weevil (Cylas formicarius); Pepper Weevil (Anthonomus eugenii); Citrus Root Weevil (Diaprepes abbreviatus); Strawberry Root Weevil (Otiorhynchus ovatus); Pecan Weevil (Curculio caryae); Filbert Weevil (Curculio occidentis); Rice Water Weevil (Lissorhoptrus oryzophilus); Alfalfa Weevil (Hypera postica); Clover Leaf Weevil (Hypera zoilus); Tea Shot-hole Borer (Euwallacea fornicatus); Sugarcane Beetle (Euetheola humilis); Coffee Berry Borer (Hypothenemus hampei); Annual Blue Grass Weevil (Listronotus maculicollis); Asiatic Garden Beetle (Maladera castanea); European Chafer (Amphimallon majale); Green June Beetle (Cotinis nitida); Japanese Beetle (Popillia japonica); May or June Beetle (Phyllophaga sp.); Northern Masked Chafer (Cyclocephala borealis); Oriental Beetle (Anomala orientalis); Southern Masked Chafer (Cyclocephala lurida); Billbug(Curculionoidea spp.); Yellow Fever Mosquito (Aedes aegypti); Maize Stalk Borer (Busseolafusca); Asiatic Rice Borer (Chilo suppressalis); Northern House Mosquito (Culex pipiens);Southern House Mosquito (Culex quinquefasciatus); Corn Rootworm (Diabrotica virgifera); Sugarcane Borer (Diatraea saccharalis); Tobacco Budworm (Heliothis virescens); Colorado Potato Beetle (Leptinotarsa decemlineata); Asian Corn Borer (Ostrinia furnacalis); European Corn Borer (Ostrinia nubilalis); Pink Bollworm (Pectinophora gossypiella); Indianmeal Moth (Plodia interpunctella); Diamondback Moth (Plutella xylostella); Soybean Looper (Chrysodeixis includens); Beet Armyworm (Spodoptera exigua); Fall Armyworm (Spodoptera frugiperda); African Cotton Leafworm (Spodoptera littoralis); Cabbage Looper (Trichoplusia ni); and Elm Leaf Beetle (Xanthogaleruca luteola).

42. A method of protecting a plant from insects comprising, providing a plant which expresses a TVP of any one of claims 1-4, or a polynucleotide encoding the same.

43. The method of claim 42, wherein the insects are selected from the group consisting of: Achema Sphinx Moth (Hornworm) (Eumorpha achemon); Alfalfa Caterpillar (Coliaseurytheme); Almond Moth (Cadra cautella); Amorbia Moth (Amorbia humerosana);Armyworm (Spodoptera spp., e.g. exigua, frugiperda, littoralis, Mythimna unipuncta); Artichoke Plume Moth (Platyptilia carduidactyla); Azalea Caterpillar (Datana major);Bagworm (Thyridopteryx ephemeraeformis); Banana Moth (Hypercompe scribonia); Banana277702-566574 Skipper (Erionota thrax); Blackheaded Budworm (Acleris gloverana); California Oakworm (Phryganidia californica); Spring Cankerworm (Paleacrita merriccata); Cherry Fruitworm (Grapholita packardi); China Mark Moth (Nymphula nitidulata); Citrus Cutworm (Egira curialis); Codling Moth (Cydia pomonella); Cranberry Fruitworm (Acrobasis vaccinii); Cross-striped Cabbageworm (Evergestis rimosalis); Cutworm (Noctuid species, Agrotis ipsilon); Douglas Fir Tussock Moth (Orgyia pseudotsugata); Ello Moth (Hornworm) (Erinnyis ello); Elm Spanworm (Ennomos subsignaria); European Grapevine Moth (Lobesia botrana); European Skipper (Thymelicus lineola); Fall Webworm (Hyphantria cunea); Filbert Leafroller (Archips rosanus); Fruittree Leafroller (Archips argyrosplia); Grape Berry Moth (Paralobesia viteana); Grape Leafroller (Platynota stultana); Grapeleaf Skeletonizer (Harrisina americana ); Green Cloverworm (Hypena scabra); Greenstriped Mapleworm (Dryocampa rubicunda); Gummosos (Batrachedra comosae); Gypsy Moth (Lymantria dispar); Hemlock Looper (Lambdina fiscellaria); Hornworm (Manduca spp.); Imported Cabbageworm (Pieris rapae); Io Moth (Automeris io); Jack Pine Budworm (Choristoneura pinus); Light Brown Apple Moth (Epiphyas postvittana); Melonworm (Diaphania hyalinata); Mimosa Webworm (Homadaula anisocentra); Obliquebanded Leafroller (Choristoneura rosaceana); Oleander Moth (Syntomeida epilais); Omnivorous Leafroller (Playnota stultana); Omnivorous Looper (Sabulodes aegrotata); Orangedog (Papilio cresphontes); Orange Tortrix (Argyrotaenia citrana); Oriental Fruit Moth (Grapholita molesta); Peach Twig Borer (Anarsia lineatella); Pine Butterfly (Neophasia menapia); Podworm (Helicoverpa zea); Redbanded Leafroller (Argyrotaenia velutinana); Redhumped Caterpillar (Schizura concinna); Rindworm Complex (Various Leps.); Saddleback Caterpillar (Acharia stimuli); Saddle Prominent Caterpillar (Heterocampa guttivitta); Saltmarsh Caterpillar (Estigmene acrea); Sod Webworm (Crambus spp.); Spanworm (Ennomos subsignaria ); Fall Cankerworm (Alsophila pometaria); Spruce Budworm (Choristoneura fumiferana); Tent Caterpillar (Various Lasiocampidae ); Fruit Borer Caterpillar (Thecla basilides); Tobacco Hornworm (Manduca sexta); Tobacco Moth (Ephestia elutella); Tufted Apple Budmoth (Platynota idaeusalis); Variegated Cutworm (Peridroma saucia); Variegated Leafroller (Platynota flavedana); Velvetbean Caterpillar (Anticarsia gemmatalis); Walnut Caterpillar (Datana integerrima); Western Tussock Moth (Orgyia vetusta); Southern Cornstalk Borer (Diatraea crambidoides); Corn Earworm (Helicoverpa armigera); Sweet Potato Weevil (Cylas formicarius); Pepper Weevil (Anthonomus eugenii); Citrus Root Weevil (Diaprepes abbreviatus); Strawberry Root Weevil (Otiorhynchus ovatus); Pecan Weevil (Curculio caryae); Filbert Weevil (Curculio occidentis); Rice Water Weevil (Lissorhoptrus277702-566574 oryzophilus); Alfalfa Weevil (Hypera postica); Clover Leaf Weevil (Hypera zoilus); Tea Shot-hole Borer (Euwallacea fornicatus); Sugarcane Beetle (Euetheola humilis); Coffee Berry Borer (Hypothenemus hampei); Annual Blue Grass Weevil (Listronotus maculicollis); Asiatic Garden Beetle (Maladera castanea); European Chafer (Amphimallon majale); Green June Beetle (Cotinis nitida); Japanese Beetle (Popillia japonica); May or June Beetle (Phyllophaga sp.); Northern Masked Chafer (Cyclocephala borealis); Oriental Beetle (Anomala orientalis); Southern Masked Chafer (Cyclocephala lurida); Billbug(Curculionoidea spp.); Yellow Fever Mosquito (Aedes aegypti); Maize Stalk Borer (Busseolafusca); Asiatic Rice Borer (Chilo suppressalis); Northern House Mosquito (Culex pipiens);Southern House Mosquito (Culex quinquefasciatus); Corn Rootworm (Diabrotica virgifera); Sugarcane Borer (Diatraea saccharalis); Tobacco Budworm (Heliothis virescens); Colorado Potato Beetle (Leptinotarsa decemlineata); Asian Corn Borer (Ostrinia furnacalis); European Corn Borer (Ostrinia nubilalis); Pink Bollworm (Pectinophora gossypiella); Indianmeal Moth (Plodia interpunctella); Diamondback Moth (Plutella xylostella); Soybean Looper (Chrysodeixis includens); Beet Armyworm (Spodoptera exigua); Fall Armyworm (Spodoptera frugiperda); African Cotton Leafworm (Spodoptera littoralis); Cabbage Looper (Trichoplusia ni); and Elm Leaf Beetle (Xanthogaleruca luteola).

44. The method of claim 43, wherein the insects are selected from the group consisting of: Aedes aegypti; Busseola fusca; Chilo suppressalis; Chrysodeixis includens; Culex pipiens; Culex quinquefasciatus; Diabrotica virgifera; Diatraea saccharalis; Helicoverpa armigera; Helicoverpa zea; Heliothis virescens; Leptinotarsa decemlineata; Ostrinia furnacalis; Ostrinia nubilalis; Pectinophora gossypiella; Plodia interpunctella; Plutella xylostella; Spodoptera exigua; Spodoptera frugiperda; Spodoptera littoralis; Trichoplusia ni; and Xanthogaleruca luteola.

45. A method of combating, controlling, or inhibiting a pest comprising: applying a pesticidally effective amount of the composition or formulation of any one of claims 11-18 to the locus of the pest, or to a plant or animal susceptible to an attack by the pest.

46. The method of claim 45, wherein the pest is selected from the group consisting of: Achema Sphinx Moth (Hornworm) (Eumorpha achemon); Alfalfa Caterpillar (Coliaseurytheme); Almond Moth (Cadra cautella); Amorbia Moth (Amorbia humerosana);Armyworm (Spodoptera spp., e.g. exigua, frugiperda, littoralis, Mythimna unipuncta);277702-566574 Artichoke Plume Moth (Platyptilia carduidactyla); Azalea Caterpillar (Datana major);Bagworm (Thyridopteryx ephemeraeformis); Banana Moth (Hypercompe scribonia); BananaSkipper (Erionota thrax); Blackheaded Budworm (Acleris gloverana); California Oakworm (Phryganidia californica); Spring Cankerworm (Paleacrita merriccata); Cherry Fruitworm (Grapholita packardi); China Mark Moth (Nymphula nitidulata); Citrus Cutworm (Egira curialis); Codling Moth (Cydia pomonella); Cranberry Fruitworm (Acrobasis vaccinii); Cross-striped Cabbageworm (Evergestis rimosalis); Cutworm (Noctuid species, Agrotis ipsilon); Douglas Fir Tussock Moth (Orgyia pseudotsugata); Ello Moth (Hornworm) (Erinnyis ello); Elm Spanworm (Ennomos subsignaria); European Grapevine Moth (Lobesia botrana); European Skipper (Thymelicus lineola); Fall Webworm (Hyphantria cunea); Filbert Leafroller (Archips rosanus); Fruittree Leafroller (Archips argyrosplia); Grape Berry Moth (Paralobesia viteana); Grape Leafroller (Platynota stultana); Grapeleaf Skeletonizer (Harrisina americana ); Green Cloverworm (Hypena scabra); Greenstriped Mapleworm (Dryocampa rubicunda); Gummosos (Batrachedra comosae); Gypsy Moth (Lymantria dispar); Hemlock Looper (Lambdina fiscellaria); Hornworm (Manduca spp.); Imported Cabbageworm (Pieris rapae); Io Moth (Automeris io); Jack Pine Budworm (Choristoneura pinus); Light Brown Apple Moth (Epiphyas postvittana); Melonworm (Diaphania hyalinata); Mimosa Webworm (Homadaula anisocentra); Obliquebanded Leafroller (Choristoneura rosaceana); Oleander Moth (Syntomeida epilais); Omnivorous Leafroller (Playnota stultana); Omnivorous Looper (Sabulodes aegrotata); Orangedog (Papilio cresphontes); Orange Tortrix (Argyrotaenia citrana); Oriental Fruit Moth (Grapholita molesta); Peach Twig Borer (Anarsia lineatella); Pine Butterfly (Neophasia menapia); Podworm (Helicoverpa zea); Redbanded Leafroller (Argyrotaenia velutinana); Redhumped Caterpillar (Schizura concinna); Rindworm Complex (Various Leps.); Saddleback Caterpillar (Acharia stimuli); Saddle Prominent Caterpillar (Heterocampa guttivitta); Saltmarsh Caterpillar (Estigmene acrea); Sod Webworm (Crambus spp.); Spanworm (Ennomos subsignaria ); Fall Cankerworm (Alsophila pometaria); Spruce Budworm (Choristoneura fumiferana); Tent Caterpillar (Various Lasiocampidae ); Fruit Borer Caterpillar (Thecla basilides); Tobacco Hornworm (Manduca sexta); Tobacco Moth (Ephestia elutella); Tufted Apple Budmoth (Platynota idaeusalis); Variegated Cutworm (Peridroma saucia); Variegated Leafroller (Platynota flavedana); Velvetbean Caterpillar (Anticarsia gemmatalis); Walnut Caterpillar (Datana integerrima); Western Tussock Moth (Orgyia vetusta); Southern Cornstalk Borer (Diatraea crambidoides); Corn Earworm (Helicoverpa armigera); Sweet Potato Weevil (Cylas formicarius); Pepper Weevil (Anthonomus eugenii); Citrus Root Weevil (Diaprepes277702-566574 abbreviatus); Strawberry Root Weevil (Otiorhynchus ovatus); Pecan Weevil (Curculio caryae); Filbert Weevil (Curculio occidentis); Rice Water Weevil (Lissorhoptrus oryzophilus); Alfalfa Weevil (Hypera postica); Clover Leaf Weevil (Hypera zoilus); Tea Shot-hole Borer (Euwallacea fornicatus); Sugarcane Beetle (Euetheola humilis); Coffee Berry Borer (Hypothenemus hampei); Annual Blue Grass Weevil (Listronotus maculicollis); Asiatic Garden Beetle (Maladera castanea); European Chafer (Amphimallon majale); Green June Beetle (Cotinis nitida); Japanese Beetle (Popillia japonica); May or June Beetle (Phyllophaga sp.); Northern Masked Chafer (Cyclocephala borealis); Oriental Beetle (Anomala orientalis); Southern Masked Chafer (Cyclocephala lurida); Billbug(Curculionoidea spp.); Yellow Fever Mosquito (Aedes aegypti); Maize Stalk Borer (Busseolafusca); Asiatic Rice Borer (Chilo suppressalis); Northern House Mosquito (Culex pipiens);Southern House Mosquito (Culex quinquefasciatus); Corn Rootworm (Diabrotica virgifera); Sugarcane Borer (Diatraea saccharalis); Tobacco Budworm (Heliothis virescens); Colorado Potato Beetle (Leptinotarsa decemlineata); Asian Corn Borer (Ostrinia furnacalis); European Corn Borer (Ostrinia nubilalis); Pink Bollworm (Pectinophora gossypiella); Indianmeal Moth (Plodia interpunctella); Diamondback Moth (Plutella xylostella); Soybean Looper (Chrysodeixis includens); Beet Armyworm (Spodoptera exigua); Fall Armyworm (Spodoptera frugiperda); African Cotton Leafworm (Spodoptera littoralis); Cabbage Looper (Trichoplusia ni); and Elm Leaf Beetle (Xanthogaleruca luteola).

47. The method of claim 45, wherein the pest is selected from the group consisting of: Aedes aegypti; Busseola fusca; Chilo suppressalis; Chrysodeixis includens; Culex pipiens; Culex quinquefasciatus; Diabrotica virgifera; Diatraea saccharalis; Helicoverpa armigera; Helicoverpa zea; Heliothis virescens; Leptinotarsa decemlineata; Ostrinia furnacalis; Ostrinia nubilalis; Pectinophora gossypiella; Plodia interpunctella; Plutella xylostella; Spodoptera exigua; Spodoptera frugiperda; Spodoptera littoralis; Trichoplusia ni; and Xanthogaleruca luteola.

48. A vector comprising a polynucleotide operable to encode a TVP having an amino acid sequence with at least 90%, or 95%, or 96%, or 97%, or 98%, or 99% or at least 100% sequence identity to an amino acid sequence as set forth in any one of SEQ ID NOs: 5-9 and 11-50, or an agriculturally acceptable salt thereof.277702-566574 49. The vector of claim 48, wherein the polynucleotide comprises a nucleotide sequence as set forth in any one of SEQ ID NOs: 51-55 and 57-96, or a complementary nucleotide sequence thereof.

50. A yeast strain comprising: a first expression cassette comprising a polynucleotide operable to encode a TVP, said TVP comprising, consisting essentially of, or consists of an amino acid sequence that is at least 90%, or 95%, or 96%, or 97%, or 98%, or 99% or at least 100% sequence identical to the amino acid sequence of SEQ ID NOs: 5-9 and 11-50; or complementary nucleotide sequence thereof.

51. The yeast strain of claim 50, wherein the yeast cell is selected from any species belonging to the genera Saccharomyces, Komagataella, Kluyveromyces, Pichia, Yarrowia or Schizosaccharomyces.

52. The yeast strain of claim 51, wherein the yeast cell is selected from the group consisting of Kluyveromyces lactis, Kluyveromyces marxianus, Saccharomyces cerevisiae, and Komagataella phaffii.

53. The method of claim 52, wherein the yeast cell is Kluyveromyces lactis or Kluyveromyces marxianus.

54. The method of claim 53, wherein the yeast cell is Kluyveromyces lactis.

55. The TVP of any one of claims 1-4, wherein the TVP consists of an amino sequence as set forth in any one of SEQ ID NOs: 5, 6, 7, or 8, or an agriculturally acceptable salt thereof.

56. The TVP of claim 55, wherein the TVP is encoded by a polynucleotide sequence as set forth in any one of SEQ ID NOs: 50, 51, 52, or 53, or a complementary nucleotide sequence thereof.

57. A composition comprising: (1) an insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), a TVP-insecticidal protein comprising two or more TVPs, or an agriculturally acceptable salt thereof; and (2) one or more agriculturally acceptable excipients; wherein the TVP or the TVP-insecticidal protein comprises an amino acid sequence that is at least 80%,277702-566574 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence as set forth in any one of SEQ ID NOs: 5-9 or 11-50; and wherein the TVP or the TVP-insecticidal protein does not consist of SEQ ID NOs: 1, 3, 4, or 10.

58. The composition of claim 57, wherein the one or more agriculturally acceptable excipients comprises a sugar, a sugar alcohol, or a combination thereof.

59. The composition of claim 58, wherein the sugar is trehalose, dextrose, lactose, galactose, maltose, lactotriose, sucrose, fructose, or glucose.

60. The composition of claim 59, wherein the sugar is maltose.

61. The composition of claim 58, wherein the sugar alcohol is erythritol, lactitol, maltitol, mannitol, or xylitol.

62. The composition of claim 61, wherein the sugar alcohol is mannitol.

63. The composition of any one of claims 57-62, wherein the TVP ranges from about 2% to about 16% w / w; and wherein the sugar or sugar alcohol ranges from about 5% to about 40% w / w; of the total weight of the composition.

64. The composition of any one of claims 57-62, wherein the one or more excipient is selected from the group consisting of: trehalose; maltodextrin; maltose; potassium phosphate dibasic (K2HPO4); potassium phosphate monobasic (KH2PO4); lignosulfonate; gypsum; sorbitol; sodium benzoate; potassium sorbate; EDTA; benzisothiazolinone (BIT); and fermentation solids.

65. The composition of any one of claims 57-62, wherein the composition comprises a TVP or a TVP-insecticidal protein, each independently in an amount ranging from about 2% to about 16% wt / wt; and wherein the trehalose ranges from about 5% to about 40% wt / wt; the BIT ranges from about 0.01% to about 0.1% wt / wt; the maltodextrin ranges from about 10% to about 50% wt / wt; the potassium phosphate dibasic anhydrous (K2HPO4) ranges from about 1% to about 5% wt / wt; the potassium phosphate monobasic (KH2PO4) ranges from about277702-566574 0.10% to about 1% wt / wt; and the fermentation solids range from about 15% to about 40% wt / wt; of the total weight of the composition.

66. The composition of any one of claims 57-62, wherein the composition comprises a TVP or a TVP-insecticidal protein, each independently ranging from about 7% to about 9% w / w; and wherein trehalose ranges from about 20% to about 30% w / w; BIT ranges from about 0.025% to about 0.075% w / w; maltodextrin ranges from about 30% to about 40% w / w; potassium phosphate dibasic anhydrous (K2HPO4) ranges from about 2% to about 3% w / w; potassium phosphate monobasic (KH2PO4) ranges from about 0.2% to about 0.6%; and fermentation solids range from about 20% to about 30%, of the total weight of the composition.

67. The composition of any one of claims 57-62, wherein the composition comprises an amount of TVP, or TVP-insecticidal protein that is 8.5% wt / wt of the total weight of the composition; and wherein the plurality of excipients consists of the following: an amount of trehalose that is 25% wt / wt; an amount of BIT that is 0.05% wt / wt; an amount of maltodextrin that is 36.3% wt / wt; an amount of potassium phosphate dibasic anhydrous (K2HPO4) that is 2.6% wt / wt; an amount of potassium phosphate monobasic (KH2PO4) that is 0.4% wt / wt; and an amount of fermentation solids that is 26.85% wt / wt, of the total weight of the composition.

68. A method of combating, controlling, or inhibiting a pest, the method comprising: applying a pesticidally effective amount of an insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP) or an agriculturally acceptable salt thereof to: the pest, a locus of the pest, a food supply of the pest, a habitat of the pest, or a breeding ground of the pest; a plant, a seed, a plant part, a locus of a plant, or an environment of a plant that is susceptible to an attack by the pest; an animal, a locus of an animal, or an environment of an animal susceptible to an attack by the pest; or a combination thereof; wherein the TVP comprises: an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a compound of Formula I: X1-X2-X3-X4-X5-C-R-A-B1-M-T- N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A-E-C-F-R-N-D-V-Y-B2-A-C-Z1-Z2- Z3-Z4-Z5-Z6 (SEQ ID NO: 98); wherein variable letters Xn and Bn and Zn are each independently amino acids in single letter format; wherein X1is E, or absent, X2is P or277702-566574 absent, X3is D or absent, X4is E, or absent, X5is I or absent, B1is R or Q, B2is T or A; Z1can be H or absent; Z2is E or absent; Z3is A or absent; Z4is Q or absent; Z5is K or absent; and Z6 is G or absent, and wherein the compound of Formula I does not consist of SEQ ID NOs: 1, 3, 4, or 10.

69. A method of combating, controlling, or inhibiting a pest, the method comprising: applying a pesticidally effective amount of an insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP) or an agriculturally acceptable salt thereof to: the pest, a locus of the pest, a food supply of the pest, a habitat of the pest, or a breeding ground of the pest; a plant, a seed, a plant part, a locus of a plant, or an environment of a plant that is susceptible to an attack by the pest; an animal, a locus of an animal, or an environment of an animal susceptible to an attack by the pest; or a combination thereof; wherein the TVP comprises: an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a compound of Formula II: X1-X2-X3-X4-X5-C-R-A-B1-M- T-N-K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A-E-C-F-R-N-D-V-Y-B2-A-C-H- E-A-Q-K-G (SEQ ID NO: 99); wherein variable letters Xn and Bn are each independently amino acids in single letter format; wherein X1is E, or absent, X2is P or absent, X3is D or absent, X4 is E, or absent, X5 is I or absent, B1 is R or Q, and B2 is T or A; and wherein the compound of Formula II does not consist of SEQ ID NOs: 1, 3, 4, or 10.

70. A method of combating, controlling, or inhibiting a pest, the method comprising: applying a pesticidally effective amount of an insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP) or an agriculturally acceptable salt thereof to: the pest, a locus of the pest, a food supply of the pest, a habitat of the pest, or a breeding ground of the pest; a plant, a seed, a plant part, a locus of a plant, or an environment of a plant that is susceptible to an attack by the pest; an animal, a locus of an animal, or an environment of an animal susceptible to an attack by the pest; or a combination thereof; wherein the TVP comprises: an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a compound of Formula III: E-P-D-E-I-C-R-A-B1-M-T-N- K-E-F-T-Y-K-S-N-V-C-N-N-C-G-D-Q-V-A-A-C-E-A-E-C-F-R-N-D-V-Y-B2-A-C-Z1-Z2- Z3-Z4-Z5-Z6 (SEQ ID NO: 100); wherein variable letters Bn and Zn are each independently amino acids in single letter format; wherein B1is R or Q, and B2is T or A; Z1is H or absent; Z2 is E or absent; Z3 is A or absent; Z4 is Q or absent; Z5 is K or absent; and Z6 is G or absent; and wherein the compound of Formula III does not consist of SEQ ID NOs: 1, 3, 4, or 10.277702-566574 71. The method of any one of claims 68-70, wherein the TVP possesses enhanced binding to one or more phospholipids of a cell membrane relative to SEQ ID NO:

4.

72. The method of any one of claims 68-70, wherein the TVP causes increased potentiation of Ach induced currents through an nAChR relative to SEQ ID NO:

4.

73. The method of any one of claims 68-72, wherein the pest is selected from the group consisting of: Acleris gloverana; Acrobasis vaccinii; Aedes aegypti; Agrotis ipsilon; Alsophila pometaria; Amorbia humerosana; Anaphothrips obscurus; Anarsia lineatella; Anomala orientalis; Anthonomus eugenii; Anticarsia gemmatalis; Archips argyrosplia; Archips rosanus; Argyrotaenia citrana; Argyrotaenia velutinana; Automeris io; Batrachedra comosae; Busseola fusca; Cadra cautella; Chilo suppressalis; Choristoneura fumiferana; Choristoneura pinus; Choristoneura rosaceana; Colias eurytheme; Cotinis nitida; Crambus spp.; Culex pipiens; Culex quinquefasciatus; Curculio caryae; Curculio occidentis; Cyclocephala borealis; Cyclocephala lurida; Cydia pomonella; Cylas formicarius; Datana integerrima; Datana major; Diabrotica virgifera; Diaphania hyalinata; Diaprepes abbreviatus; Diatraea saccharalis; Dryocampa rubicunda; Ennomos subsignaria; Ephestia elutella; Epiphyas postvittana; Erinnyis ello; Erionota thrax; Estigmene acrea; Euetheola humilis; Eumorpha achemon; Euwallacea fornicatus; Evergestis rimosalis; Frankliniella occidentalis; Frankliniella fusca; Frankliniella intonsa; Frankliniella schultzei; Grapholita molesta; Grapholita packardi; Harrisina americana; Helicoverpa armigera; Helicoverpa zea; Heliothis virescens; Hercinothrips femoralis; Cecrita guttivitta; Homadaula anisocentra; Hypera postica; Brachypera zoilus; Hypercompe scribonia; Hyphantria cunea; Hypothenemus hampei; Lambdina fiscellaria; Lasiocampidae spp.; Leptinotarsa decemlineata; Lissorhoptrus oryzophilus; Listronotus maculicollis; Lobesia botrana; Lymantria dispar; Maladera castanea; Manduca quinquemaculata; Manduca sexta; Cydia latiferreana; Neohydatothrips variabilis; Neophasia menapia; Noctuid spp.; Nymphula nitidulata; Orgyia pseudotsugata; Orgyia vetusta; Ostrinia furnacalis; Ostrinia nubilalis; Otiorhynchus ovatus; Paleacrita merriccata; Papilio cresphontes; Paralobesia viteana; Pectinophora gossypiella; Peridroma saucia; Pezothrips kellyanus; Phryganidia californica; Phyllophaga spp.; Pieris rapae; Hypena scabra; Platynota flavedana; Platynota idaeusalis; Platynota stultana; Platyptilia carduidactyla; Plodia interpunctella; Plutella xylostella; Popillia japonica; Pseudaletia unipuncta; Chrysodeixis includens; Amphimallon majale;277702-566574 Sabulodes aegrotata; Schizura concinna; Scirtothrips perseae; Neohydatothrips variabilis; Acharia stimuli; Diatraea crambidoides; Sphenophorus apicalis; Sphenophorus cariosus; Sphenophorus inaequalis; Sphenophorus minimus; Sphenophorus parvulus; Spodoptera exigua; Spodoptera frugiperda; Spodoptera littoralis; Syntomeida epilais; Thecla basilides; Thrips palmi; Thrips tabaci; Thymelicus lineola; Thyridopteryx ephemeraeformis; Trichoplusia ni; Xanthogaleruca luteola; and Egira curialis.

74. A plant, plant part thereof, comprising a leaf, a stem a flower, a sepal, a fruit, a nut, a root, a seed, or combinations thereof, comprising a TVP having an amino acid sequence with at least 90%, or 95%, or 96%, or 97%, or 98%, or 99% or at least 100% sequence identity to an amino acid sequence as set forth in any one of SEQ ID NOs: 5-9 and 11-50, or having the composition or formulation according to any one of claims 11-18, or 57-67, applied thereto.

75. The plant or plant part thereof, of claim 74, wherein the plant or plant part comprises a plant or plat part selected from: alfalfa, cotton, tomato, maize, wheat, corn, sweet corn, lucerne, soybean, sorghum, field pea, linseed, safflower, rapeseed, oil seed rape, rice, soybean, barley, sunflower, trees (including coniferous and deciduous), flowers (including those grown commercially and in greenhouses), field lupins, switchgrass, sugarcane, potatoes, tobacco, crucifers, peppers, sugarbeet, barley, and oilseed rape, Brassica sp., rye, millet, peanuts, sweet potato, cassaya, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana, avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond, oats, vegetables, ornamentals, and conifers.

76. A method for delivering a composition to an insect, the method comprising applying the composition according to any one of claims 11-18 or 57-67 to the plant or plant part thereof, to obtain a treated material suitable for contacting of, or ingestion by insects.

77. A method for controlling insect infestation and / or damage by insect pests, comprising applying a composition according to any one of claims 11-18 or 57-67 to an insect or its environment, a plant, or plant part thereof, or the locus where the plant or plant part thereof is planted.277702-566574 78. The method according to claim 77, wherein the composition comprises a TVP or a TVP-insecticidal protein, both individually having insecticidal activity when ingested by an insect.

79. Use of a composition according to any of claims 11-18 or 57-67 for controlling a pest infestation.

80. The use of claim 79, wherein the pest is selected from the group consisting of: Acleris gloverana; Acrobasis vaccinii; Aedes aegypti; Agrotis ipsilon; Alsophila pometaria; Amorbia humerosana; Anaphothrips obscurus; Anarsia lineatella; Anomala orientalis; Anthonomus eugenii; Anticarsia gemmatalis; Archips argyrosplia; Archips rosanus; Argyrotaenia citrana; Argyrotaenia velutinana; Automeris io; Batrachedra comosae; Busseola fusca; Cadra cautella; Chilo suppressalis; Choristoneura fumiferana; Choristoneura pinus; Choristoneura rosaceana; Colias eurytheme; Cotinis nitida; Crambus spp.; Culex pipiens; Culex quinquefasciatus; Curculio caryae; Curculio occidentis; Cyclocephala borealis; Cyclocephala lurida; Cydia pomonella; Cylas formicarius; Datana integerrima; Datana major; Diabrotica virgifera; Diaphania hyalinata; Diaprepes abbreviatus; Diatraea saccharalis; Dryocampa rubicunda; Ennomos subsignaria; Ephestia elutella; Epiphyas postvittana; Erinnyis ello; Erionota thrax; Estigmene acrea; Euetheola humilis; Eumorpha achemon; Euwallacea fornicatus; Evergestis rimosalis; Frankliniella occidentalis; Frankliniella fusca; Frankliniella intonsa; Frankliniella schultzei; Grapholita molesta; Grapholita packardi; Harrisina americana; Helicoverpa armigera; Helicoverpa zea; Heliothis virescens; Hercinothrips femoralis; Cecrita guttivitta; Homadaula anisocentra; Hypera postica; Brachypera zoilus; Hypercompe scribonia; Hyphantria cunea; Hypothenemus hampei; Lambdina fiscellaria; Lasiocampidae spp.; Leptinotarsa decemlineata; Lissorhoptrus oryzophilus; Listronotus maculicollis; Lobesia botrana; Lymantria dispar; Maladera castanea; Manduca quinquemaculata; Manduca sexta; Cydia latiferreana; Neohydatothrips variabilis; Neophasia menapia; Noctuid spp.; Nymphula nitidulata; Orgyia pseudotsugata; Orgyia vetusta; Ostrinia furnacalis; Ostrinia nubilalis; Otiorhynchus ovatus; Paleacrita merriccata; Papilio cresphontes; Paralobesia viteana; Pectinophora gossypiella; Peridroma saucia; Pezothrips kellyanus; Phryganidia californica; Phyllophaga spp.; Pieris rapae; Hypena scabra; Platynota flavedana; Platynota idaeusalis; Platynota stultana; Platyptilia carduidactyla; Plodia interpunctella; Plutella xylostella; Popillia japonica; Pseudaletia unipuncta; Chrysodeixis includens; Amphimallon majale;277702-566574 Sabulodes aegrotata; Schizura concinna; Scirtothrips perseae; Neohydatothrips variabilis; Acharia stimuli; Diatraea crambidoides; Sphenophorus apicalis; Sphenophorus cariosus; Sphenophorus inaequalis; Sphenophorus minimus; Sphenophorus parvulus; Spodoptera exigua; Spodoptera frugiperda; Spodoptera littoralis; Syntomeida epilais; Thecla basilides; Thrips palmi; Thrips tabaci; Thymelicus lineola; Thyridopteryx ephemeraeformis; Trichoplusia ni; Xanthogaleruca luteola; and Egira curialis.

81. A polynucleotide operable to hybridize under stringent hybridization conditions with a polynucleotide segment encoding the TVP of any one of claims 1-4.

82. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP consisting of an amino acid sequence of SEQ ID NO:

5.

83. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP consisting of an amino acid sequence of SEQ ID NO:

6.

84. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP consisting of an amino acid sequence of SEQ ID NO:

7.

85. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP consisting of an amino acid sequence of SEQ ID NO:

8.

86. An insecticidal U1-agatoxin-Ta1b variant polypeptide (TVP), said TVP consisting of an amino acid sequence of SEQ ID NO: 9.

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