Bacterial delivery vehicles containing tracer nucleic acid sequences

Abstract

The present disclosure generally relates to genetically tagged bacterial delivery vehicles comprising unique tracer nucleic acid sequences (herein referred to as "tracers") for use in detecting and / or quantifying the presence of two or more different bacterial delivery vehicles in a mixture of vehicles. The present disclosure relates to methods in which bacterial delivery vehicles are detected, for example, by performing multiple cycles of amplification using primers that bind to sequences within the unique tracers. Such methods can be advantageously used in quality control to detect and quantify mixtures of bacterial delivery vehicles within pharmaceutical compositions.

Description

[Technical Field]

[0001] The present disclosure generally relates to genetically tagged bacterial delivery vehicles comprising unique tracer nucleic acid sequences (herein referred to as "tracers") for use in detecting and / or quantifying the presence of two or more different bacterial delivery vehicles in a mixture of vehicles. The present disclosure relates to methods in which the bacterial delivery vehicles are detected, for example, by performing multiple cycles of amplification using primers that bind to sequences within the unique tracers. Such methods can be advantageously used in quality control to detect and quantify mixtures of bacterial delivery vehicles within pharmaceutical compositions. [Background technology]

[0002] Bacterial delivery vehicles, such as packaged phagemids, are bacteriophage-derived particles composed of a desired DNA nucleic acid payload packaged in a bacteriophage-derived capsid. Cellular delivery vehicles, in which the capsid is the primary host range determinant, follow the same host range restrictions as bacteriophages, allowing different bacterial delivery vehicles with complementary host ranges to be combined in a multivalent final drug product (a drug product with multiple target cell specificities) to achieve clinical efficacy. Therefore, approaches using bacteriophage-based bacterial delivery vehicles for use as antibacterial drugs may rely on cocktail formulations, in which multiple phages with different host ranges are combined and administered to a subject as a mixture to maximize clinical efficacy by targeting the greatest number of clinically relevant bacterial strains. In such cases, different bacterial delivery vehicles in the final composition mixture contain the same or different DNA but have different capsids with different host ranges and / or binding specificities (see Figure 1).

[0003] From a regulatory perspective, it is necessary to characterize the composition of a drug product before administering it to a subject. Particularly for drug products that are composed of a mixture of different drug substances, the presence and amount of each drug substance in the final product must be confirmed. For example, the presence and amount of each bacterial delivery vehicle in a pharmaceutical composition for use in treating a subject must be evaluated in the final product, and it must comply with the specifications set for the drug. Therefore, a method is required to detect the presence and amount of different bacterial delivery vehicles in a pharmaceutical composition for regulatory purposes. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] U.S. Provisional Patent Application No. 62 / 849,108 [Patent Document 2] U.S. Provisional Patent Application No. 62 / 849,112 [Patent Document 3] U.S. Provisional Patent Application No. 62 / 802,777 [Patent Document 4] U.S. Provisional Patent Application No. 62 / 771,761 [Patent Document 5] U.S. Provisional Patent Application No. 62 / 783,258 [Non-patent literature]

[0005] [Non-Patent Document 1] Krupovic et al., Arch Virol, 2015 [Non-patent document 2] Kues, U and Stahl, U, 1989, Microbiol Rev 53:491-516. [Non-patent document 3] Del Solar et al., 1998, Microhio and Molec Biol. Rev 62:434-464. [Non-patent document 4] Taylor and Harris, 2012, Mol Ecol Resour. 2012 May;12(3):377~88 [Non-patent document 5] Kress et al., 2015, Trends Ecol Evol, Jan; 30(1): pp. 25-35. [Non-patent document 6] Liszczak and Muir, 2019, Angew Chem Int Ed Engl, Mar 22;58(13):4144~4162 [Non-Patent Document 7] Mohammadi-Kambs M. et al., 2017, ACS Omega. 2017 Apr 30; 2(4): 1302-1308 Summary of the Invention [Problem to be solved by the invention]

[0006] The present disclosure generally relates to methods for genetically tagging bacterial delivery vehicles for use in detecting the presence of two or more different (e.g., structurally distinct) bacterial delivery vehicles (herein referred to as a "polyvalent mixture of bacterial delivery vehicles") in a sample. Each of the different bacterial delivery vehicles may differ, for example, by their distinct bacterial cell binding ability and / or their distinct host range, and / or by comprising a different nucleic acid payload within the vehicle. More specifically, the present disclosure relates to methods in which the detected bacterial delivery vehicle comprises a desired nucleic acid payload that additionally contains a unique tracer that can be detected, for example, by performing cycles of amplification. Such methods can be advantageously used in the quality control of pharmaceutical compositions comprising a polyvalent bacterial delivery vehicle mixture. [Means for solving the problem]

[0007] In one embodiment, a bacterial delivery vehicle is provided that is engineered to bind to a target cell, e.g., a bacterial cell, and includes a nucleic acid payload with a unique tracer embedded within the nucleic acid payload. In another embodiment, a multivalent mixture of bacterial delivery vehicles is provided that includes two or more different bacterial delivery vehicles, each of which includes a nucleic acid payload with a unique tracer nucleic acid sequence.

[0008] In one embodiment, each of the different bacterial delivery vehicles in the multivalent mixture contains the same nucleic acid payload, except that it has a unique tracer associated with the same payload. In another embodiment, each of the different bacterial delivery vehicles in the multivalent mixture contains a different nucleic acid payload, with a unique tracer associated with each of the different payloads. Linking the unique tracer to the packaged nucleic acid payload allows for the identification and quantification of related bacterial delivery vehicles (referred to herein as "homologous bacterial delivery vehicles") packaged with tagged DNA nucleic acid payloads.

[0009] In some embodiments, the tracer comprises a stretch of homology of 20 nucleotides or less with the DNA of the bacterial production strain and / or the DNA of the target bacterial cell.

[0010] Preferably, the tracer comprises a barcode. In particular, the tracer may comprise a constant region and a barcode. In particular, the tracer may comprise a barcode flanked on both sides by constant regions. The barcode and / or constant region may be between 25 and 50 nucleic acids in length.

[0011] The tracer may be embedded in a non-coding region or in a coding region. In embodiments where the tracer is embedded in a coding region, the tracer may contain altered codon usage while encoding a protein with an unaltered amino acid sequence.

[0012] In some embodiments, the bacterial delivery vehicle contained in the multivalent mixture is a bacteriophage-derived scaffold.

[0013] Methods are provided for detecting and / or quantifying bacterial delivery vehicles in a multivalent mixture of bacterial delivery vehicles by detecting a unique tracer that correlates with the presence of a particular bacterial delivery vehicle. The methods include detecting and optionally quantifying each bacterial delivery vehicle in the multivalent mixture of bacterial delivery vehicles by performing cycles of amplification using primers that bind to nucleic acid sequences within the tracer sequence. Alternatively or additionally, the methods may include detecting and optionally quantifying all bacterial delivery vehicles in the multivalent mixture of bacterial delivery vehicles by performing cycles of amplification using primers that bind to nucleic acid sequences within the tracer sequence. Such amplification methods include, for example, PCR, qPCR, ddPCR, LCR, FISH, or NGS.

[0014] Also provided is a method for detecting and tracking bacterial delivery vehicles after administering a polyvalent mixture of bacterial delivery vehicles to a subject, wherein each bacterial delivery vehicle comprises a nucleic acid payload with a unique tracer. The method includes detecting and quantifying each different bacterial delivery vehicle in a sample from the subject, for example, by performing cycles of amplification using primers that bind to nucleic acid sequences in the tracer. Amplification of the specific tracer sequence is then correlated with the presence of the specific bacterial delivery vehicle in the subject sample. Alternatively or additionally, the method may include detecting and optionally quantifying all of the bacterial delivery vehicles in the sample from the subject by performing multiple cycles of amplification using primers that bind to the unique tracer nucleic acid sequences. Such amplification methods include, for example, PCR, qPCR, ddPCR, or NGS.

[0015] In the methods of the present invention, the unique tracer may have a constant region to which a primer can bind for initiation of an amplification reaction, and thus the method may include detecting and optionally quantifying each bacterial delivery vehicle by amplification of the tracer using a primer that binds within the constant region of the tracer sequence. Optionally, the unique tracer may further include a variable sequence to which a primer can bind for amplification, and the method may further include a separate or second amplification reaction using a primer that binds to the variable sequence for the amplification method.

[0016] The unique tracer may include a variable region to which a primer can bind for amplification, and thus the method may include detecting and optionally quantifying each bacterial delivery vehicle by amplification of the tracer using primers that bind within the variable region of the tracer sequence.

[0017] In the methods of the present invention, each bacterial delivery vehicle may contain a nucleic acid payload with the same sequence except for the tracer, or each bacterial delivery vehicle may contain a nucleic acid payload with a different sequence and a different tracer associated with each different payload.

[0018] Preferably, the tracer contains a stretch of homology of no more than 20 nucleotides with the DNA of the bacterial production strain and / or the DNA of the target bacterial cell.

[0019] In the methods of the present invention, the tracer may comprise a barcode. In particular, the tracer may comprise a constant region and a barcode, more particularly, a barcode flanked on both sides by constant regions. Preferably, the barcode and / or the constant region are between 25 and 50 nucleic acids in length.

[0020] In the methods of the present invention, the tracer may be embedded in a non-coding region or a coding region. In embodiments where the tracer is embedded in a coding region, the tracer may encode a protein that may contain altered codon usage but have unchanged amino acids.

[0021] The present invention also provides bacterial delivery vehicles as defined above, i.e. comprising a nucleic acid payload having a tracer nucleic acid sequence, as well as pharmaceutical compositions comprising the multivalent mixtures of the present invention.

[0022] The present invention also relates to a bacterial delivery vehicle or pharmaceutical composition according to the present invention for use in the treatment of a disease or disorder caused by a bacterium, preferably selected from the group consisting of infectious diseases caused by bacteria, metabolic disorders such as obesity and diabetes, and pathologies involving bacteria of the human microbiome.

[0023] For a better understanding of the subject matter disclosed herein, and to illustrate how it may be carried out in practice, embodiments will now be described, by way of non-limiting example, with reference to the accompanying drawings, in which: With particular reference to the drawings, it is emphasized that the features shown are by way of example and for purposes of illustration of embodiments of the invention. [Brief explanation of the drawings]

[0024] [Figure 1] The advantages of a mixture of phagemid particles containing a unique DNA tracer within its DNA nucleic acid payload in the context of pharmaceutical applications are demonstrated. [Figure 2] 1 illustrates the design and implementation of DNA tracers within non-coding DNA sequences of DNA nucleic acid payloads. [Figure 3] This represents the design and implementation of DNA within the synonymous coding DNA sequence of a DNA nucleic acid payload. [Figure 4] A representative example of codon possibilities for the 10 amino acid segment AA40-49 in the example of TEM-1 β-lactamase is shown. [Figure 5] Schematic representation of the plasmid backbone pAK272B, the location of the barcode region, and the corresponding primers. [Figure 6]The numbers represent the number of copies of each plasmid detected per μl from a mixture of different plasmids containing each different barcode. [Figure 7] The number of copies of each plasmid detected in the plasmid mixture extracted from in vivo transduced bacteria is shown. DETAILED DESCRIPTION OF THE INVENTION

[0025] Detailed Description The present disclosure relates generally to methods for genetically tagging bacterial delivery vehicles with desired structural features for use in detecting the presence of two or more different bacterial delivery vehicles in a polyvalent mixture of delivery vehicles. More particularly, the disclosure relates to methods in which the detected bacterial delivery vehicles contain a desired nucleic acid payload of interest that additionally contains a unique tracer nucleotide sequence that can be detected by performing cycles of amplification.

[0026] In one aspect, bacterial delivery vehicles are provided that have desired structural characteristics, such as specific target cell binding and / or host range, and contain a nucleic acid payload with an embedded unique tracer nucleic acid sequence. In another aspect, multivalent mixtures of bacterial delivery vehicles are provided that contain two or more different bacterial delivery vehicles, each containing a nucleic acid payload with a unique tracer nucleic acid sequence.

[0027] The bacterial delivery vehicles provided herein enable the transfer of a nucleic acid payload encoding a protein or nucleic acid of interest into a desired target bacterial host cell. The bacterium targeted by the bacterial delivery vehicle can be any bacterium present in a mammalian organism. It can be any commensal, symbiotic, or pathogenic bacterium of the microbiota or microbiome. The microbiota can include various bacterial species, any of which can be targeted by the present disclosure. In some embodiments, the genus and / or species of the target bacterial cell can vary depending on the type of bacteriophage used to prepare the bacterial delivery vehicle. For example, some bacteriophages exhibit tropism or preferentially target specific host species of bacteria. Other bacteriophages do not exhibit such tropism and can be used to target several different genera and / or species of endogenous bacterial cells. In a preferred embodiment, the target bacterial host cell is a cell of the microbiome. In more preferred embodiments, the target bacterial host cells are cells of the skin microbiome, the gut microbiome, the lung microbiome, or the oral microbiome.

[0028] As used herein, the term "delivery vehicle" refers to any means that allows the transfer of a nucleic acid payload into bacteria. Several types of delivery vehicles are encompassed by the present invention, including, but not limited to, bacteriophage scaffolds, viral scaffolds, chemical-based delivery vehicles (e.g., cyclodextrin, calcium phosphate, cationic polymers, cationic liposomes), protein- or peptide-based delivery vehicles, lipid-based delivery vehicles, nanoparticle-based delivery vehicles, non-chemical-based delivery vehicles (e.g., transformation, electroporation, sonication, optical transfection), particle-based delivery vehicles (e.g., gene guns, magnetofection, impale infection, biolistics, cell-penetrating peptides), or donor bacteria (conjugation). Any combination of delivery vehicles is also encompassed by the present invention. A delivery vehicle can refer to a bacteriophage-derived scaffold and can be derived from a natural capsid, an evolved capsid, or an engineered capsid. In some embodiments, the delivery vehicle is a nucleic acid payload, such that bacteria are naturally competent to take up nucleic acid payloads from the environment or themselves. In some embodiments, the bacterial delivery vehicle is a bacteriophage-derived particle comprised of a DNA nucleic acid payload of interest packaged in a bacteriophage-derived capsid.

[0029] Bacteriophage-derived particles can be prepared from bacterial viruses selected so that they are capable of introducing a nucleic acid payload into target bacteria.

[0030] The bacterial virus is preferably a bacteriophage. Bacteriophages are obligate intracellular parasites that replicate within cells by utilizing some or all of the host's biosynthetic machinery. Phage genomes come in a variety of sizes and shapes (e.g., linear or circular). Most phages range in size from 24 to 200 nm in diameter. Phages contain nucleic acid (i.e., genome) and protein and may be surrounded by a lipid membrane. Depending on the phage, the nucleic acid genome may be DNA or RNA and may exist in a circular or linear shape. The size of the phage genome varies among phages. The simplest phages have genomes only a few thousand nucleotides in size, but more complex phages may contain more than 100,000 nucleotides, and in rare cases, more than 1,000,000 nucleotides, within their genomes. The number and amount of individual types of proteins within a phage particle will vary among phages.

[0031] Optionally, the bacteriophage is selected from the Order Caudovirales, based on the taxonomy of Krupovic et al., Arch Virol, 2015: - the Myoviridae family (for example, but not limited to, the genus Cp220virus, the genus Cp8virus, the genus Ea214virus, the genus Felixo1virus, the genus Mooglevirus, the genus Suspvirus, the genus Hp1virus, the genus P2virus, the genus Kayvirus, the genus P100virus, the genus Silviavirus, the genus Spo1virus, the genus Tsarbombavirus, the genus Twortvirus, the genus Cc31virus, the genus Jd18virus, the genus Js98virus, the genus Kp15virus, the genus Moonvirus, the genus Rb49virus, the genus Rb69virus, the genus S16virus, the genus Schizot4virus, the genus Sp18virus, the genus T4virus, the genus Cr3virus, the genus Se1virus, the genus V5virus, the genus Abouovirus, the genus Agatevirus, the genus Agrican357virus, the genus Ap22virus, the genus Arv1virus, the genus B4virus) , Bastillevirus genus, Bc431virus genus, Bcep78virus genus, Bcepmuvirus genus, Biquartavirus genus, Bxz1virus genus, Cd119virus genus, Cp51virus genus, Cvm10virus genus, Eah2virus genus, Elvirus genus, Hapunavirus genus, Jimmervirus genus, Kpp10virus genus, M12virus genus, Machinavirus genus, Marthavirus genus, Msw3virus genus, Muvirus genus, Myohalovirus genus, Nit1virus genus, P1virus genus, Pakpunavirus genus, Pbunavirus genus, Phikzvirus genus, Rheph4virus genus, Rsl2virus genus, Rslunavirus genus, Secunda5virus genus, Sep1virus genus, Spn3virus genus, Svunavirus genus, Tg1virus genus, Vhmlvirus genus and Wphvirus genus). - the family Podoviridae (for example, but not limited to, the genus Fri1virus, the genus Kp32virus, the genus Kp34virus, the genus Phikmvvirus, the genus Pradovirus, the genus Sp6virus, the genus T7virus, the genus Cp1virus, the genus P68virus, the genus Phi29virus, the genus Nona33virus, the genus Pocjvirus, the genus Tl2011virus, the genus Bcep22virus, the genus Bpp1virus, the genus Cba41virus, the genus Dfl12virus, Lus virus genus, Ea92 virus genus, Epsilon15 virus genus, F116 virus genus, G7c virus genus, Jwalpha virus genus, Kf1 virus genus, Kpp25 virus genus, Lit1 virus genus, Luz24 virus genus, Luz7 virus genus, N4 virus genus, Nonana virus genus, P22 virus genus, Page virus genus, Phieco32 virus genus, Prtb virus genus, Sp58 virus genus, Una961 virus genus and Vp5 virus genus). - the family Siphoviridae (for example, but not limited to, the genera Camvirus, Likavirus, R4virus, Acadianvirus, Coopervirus, Pg1virus, Pipefishvirus, Rosebushvirus, Brujitavirus, Che9cvirus, Hawkeyevirus, Plotvirus, Jerseyvirus, K1gvirus, Sp31virus, Lmd1virus, Una4virus, Bongovirus, Reyvirus, But ters virus genus, Charlie virus genus, Redi virus genus, Baxter virus genus, Nymphadora virus genus, Bignus virus genus, Fishburne virus genus, Phayonce virus genus, Kp36 virus genus, Rogue1 virus genus, Rtp virus genus, T1 virus genus, Tls virus genus, Ab18 virus genus, Amigo virus genus, Anatole virus genus, Andromeda virus genus, Attis virus genus, Barnyard virus genus, Bernal13 virus genus, Biseptima virus Virus genus, Bronvirus genus, C2virus genus, C5virus genus, Cba181virus genus, Cbastvirus genus, Cecivirus genus, Che8virus genus, Chivirus genus, Cjw1virus genus, Corndogvirus genus, Cronusvirus genus, D3112virus genus, D3virus genus, Decurrovirus genus, Demosthenesvirus genus, Doucettevirus genus, E125virus genus, Eiauvirus genus, Ff47virus genus, Gaiavirus genus, Gilesvirus genus, Gordonvirus genus, Gord tnk virus genus, Harrison virus genus, Hk578 virus genus, Hk97 virus genus, Jenst virus genus, Jwx virus genus, Kellezio virus genus, Korra virus genus, L5 virus genus, Lambda virus genus, Laroye virus genus, Liefie virus genus, Marvin virus genus, Mudcat virus genus, N15 virus genus, Nonag virus genus, Np1 virus genus, Omega virus genus, P12002 virus genus, P12024 virus genus, P23 virus genus, P70 virus genus, Pa6 virus genus,Pamx74 virus genus, Patience virus genus, Pbi1 virus genus, Pepy6 virus genus, Pfr1 virus genus, Phic31 virus genus, Phicbk virus genus, Phieta virus genus, Phifel virus genus, Phijl1 virus genus, Pis4a virus genus, Psa virus genus, Psimuna virus genus, Rdjl virus genus, Rer2 virus genus, Sap6 virus genus, Send513 virus genus, Septima3 virus genus, Seurat virus genus, Sextaec virus genus, Sfi11 virus genus, Sfi21dt1 virus genus, Sitara virus genus , Sk1virus genus, Slashvirus genus, Smoothievirus genus, Soupsvirus genus, Spbetavirus genus, Ssp2virus genus, T5virus genus, Tankvirus genus, Tin2virus genus, Titanvirus genus, Tm4virus genus, Tp21virus genus, Tp84virus genus, Triavirus genus, Trigintaduovirus genus, Vegasvirus genus, Vendettavirus genus, Wbetavirus genus, Wildcatvirus genus, Wizardvirus genus, Woesvirus genus, Xp10virus genus, Ydn12virus genus and Yuavirus genera), and - Bacteriophages from the family Ackermannviridae (for example, but not limited to, the genera Ag3, Limestone, Cba120 and Vi1).

[0032] In some cases, the bacteriophage is not part of the Caudovirales order, but belongs to a family with an unclassified order, such as, but not limited to, the family Tectiviridae (e.g., genera Alphatectiviridae, Betatectiviridae), the family Corticoviridae (e.g., genus Corticovirus), the family Inoviridae (e.g., genus Fibrovirus, genus Habenivirus, genus Inovirus, genus Lineavirus, genus Plectrovirus, genus Saevirus, genus tivirus, Vespertiliovirus), Cystoviridae (e.g., Cystovirus), Leviviridae (e.g., Allolevivirus, Levivirus), Microviridae (e.g., Alpha3microvirus, G4microvirus, Phix174microvirus, Bdellomicrovirus, Chlamydiamicrovirus, Spiromicrovirus), and Plasmaviridae (e.g., Plasmavirus).

[0033] In some cases, the bacteriophage targets archaea and is not part of the Caudovirales but is from a family with an unclassified order, such as, but not limited to, Ampullaviridae, Fuselloviridae, Globuloviridae, Guttaviridae, Liposthrixviridae, Pleolipoviridae, Rudiviridae, Salterprovirus, and Bicaudaviridae.

[0034] A non-exhaustive list of bacterial genera and their known host-specific bacterial viruses is presented in the following paragraphs. Synonyms and orthographic variations are indicated in parentheses. Homophones are repeated the number of times they occur (e.g., D, D, d). Unnamed phages are indicated by "NN" next to their genus and their number in parentheses.

[0035] Bacteria of the genus Actinomyces can be infected by the following phages: Av-I, Av-2, Av-3, BF307, CT1, CT2, CT3, CT4, CT6, CT7, CT8 and 1281.

[0036] Bacteria of the genus Aeromonas can be infected by the following phages: AA-I, Aeh2, N, PMl, TP446, 3, 4, 11, 13, 29, 31, 32, 37, 43, 43-10T, 51, 54, 55R.1, 56, 56RR2, 57, 58, 59.1, 60, 63, Aehl, F, PM2, 1, 25, 31, 40RR2.8t, (syn= 44R), (syn= 44RR2.8t), 65, PM3, PM4, PM5, and PM6.

[0037] A、aizl、Al-KI、B 、BCJAl、BCl、BC2、BLLl、BLl、BP142、BSLl、BSL2、BS l、BS3、BS8、BS15、BS18、BS22、BS26、BS28、BS31、BS104、BS105、BS106、BTB、B1715V1、C、CK-I、Coll、Co rl, CP-53, CS-I, CSi, D, D, D, D5, entl, FP8, FP9, FSi, FS2, FS3, FS5, FS8, FS9, G, GH8, GT8, GV-I, GV-2, GT-4, g3, gl2, gl3, gl4, gl6, gl7, g21, g23, g24, g29, H2, kenl, KK-88, Kuml, Kyul, J7W-1, LP52, (syn= LP-52)、L7、Mexl、MJ-I、mor2、MP-7、MPlO、MP12、MP14、MP15、Neol、N° 2、N5、N6P、PBCl、PBLA、PBPl、P2、Sa、SF2、SF6、Shal、Sill、SP02、(syn= ΦSPP1)、SPβ、STI、STi、SU-Il、t、TbI、Tb2、Tb5、TbIO、Tb26、Tb51、Tb53、Tb55、 Tb77、Tb97、Tb99、Tb560、Tb595、Td8、Td6、Tdl5、TgI、Tg4、Tg6、Tg7、Tg9、TgIO、 TgIl、Tgl3、Tgl5、Tg21、Tinl、Tin7、Tin8、Tinl3、Tm3、Tocl、Togl、toll、TP-I 、TP-10vir、TP-15c、TP-16c、TP-17c、TP-19、TP35、TP51、TP-84、Tt4、Tt6、type A、type B、type C、type D、type E、Tφ3、VA-9、W、wx23、wx26、Yunl、α、γ、pl l、φmed-2、φT、φμ-4、φ3T、φ75、φlO5、(syn= φlO5)、IA、IB、1-97A、1-97B、2、2、3、3、3、5、12、14、20、30、35、36、37、38、41C、51、6 3、64、138D、I、II、IV、NN-バチルス(13)、alel、ARl、AR2、AR3、AR7、AR9、Bace-11、(syn= 11)、Bastille、BLl、BL2、BL3、BL4、BL5、BL6、BL8、BL9、BP124. BS28, BS80, Ch, CP-51, CP-54, D-5, Darl, Denl, DP-7, Entl, FoSi, FoS2, FS4, FS6, FS7, G, Gall, Gamma, G El, GF-2, GSi, GT-I, GT-2, GT-3, GT-4, GT-5, GT-6, GT-7, GV-6, gl5, 19, 110, ISi, K, MP9, MP13, MP21, MP23 MP24, MP28, MP29, MP30, MP32, MP34, MP36, MP37, MP39, MP40, MP41, MP43, MP44, MP45, MP47, MP50, NLP-I No .l, N17, N19, PBSl, PKl, PMBl, PMB12, PMJl, S, SPOl, SP3, SP5, SP6, SP7, SP8, SP9, SPlO, SP-15, SP50 (syn= SP-50) SP82 SST subl SW Tg8 Tgl2 Tgl3 Tgl4 thul thuΛ thuS Tin4 Tin23 TP-13 TP33 TP50 TSP-I type V type VI, V, Vx, β22, φe, φNR2, φ25, φ63, 1, 1, 2, 2C, 3NT, 4, 5, 6 7, 8, 9, 10, 12, 12, 17, 18, 19, 21, 138, III, 4(B. megateriwn))、4(B.セァリス(B. sphaericus)) AR13 BPP-IO BS32 BS107 Bl B2 GA-I GP-IO GV-3 GV-5 g8 MP20 MP27 MP49N f PP5, PP6, SF5, Tgl8, TP-I, Versailles, φl5, φ29, 1-97, 837 / IV, mι-バチス(1), BatlO, BSLlO, BSLI l, BS6, BSI l, BS16, BS23, BSlOl, BS102, gl8, morl, PBLl, SN45, thu2, thu3, TmI, Tm2, ​​TP-20, TP21, TP52, type F, type G, type IV, HN-BacMus(3), BLE (syn= θc) BS2, BS4, BS5, BS7, BlO, B12, BS20, BS21, F, MJ-4, PBA12. AP50, AP50-04, AP50-11, AP50-23, AP50-26, AP50-27, and Bam3The following Bacillus-specific phages have been deleted: DLP10716, DLP-11946, DPB5, DPB12, DPB21, DPB22, DPB23, GA-2, M, No. IM, PBLB, PBSH, PBSV, PBSW, PBSX, PBSY, PBSZ, phi, SPa, type 1, and μ.

[0038] Bacteria of the genus Bacteriodes can be infected by the following phages: crAss-phage, ad I2, Baf-44, Baf-48B, Baf-64, Bf-I, Bf-52, B40-8, F1, β1, φAl, φBrOl, φBrO2, 11, 67.1, 67.3, 68.1, mt-Bacteroides (3), Bf42, Bf71, HN-Bdellovibrio (1), and BF-41.

[0039] Bacteria of the genus Bordetella can be infected by the following phages: 134 and NN-Bordetella (3).

[0040] Bacteria of the genus Borrellia can be infected by the following phages: NN-Borrellia (1) and NN-Borrellia (2).

[0041] Brucella bacteria can be infected by the following phages: A422, Bk, (syn= Berkeley), BM29, FOi, (syn= FOl), (syn= FQl), D, FP2, (syn= FP2), (syn= FD2), Fz, (syn= Fz75 / 13), (syn= Firenze 75 / 13), (syn= Fi), Fi, (syn= Fl), Fim, (syn= FIm), (syn= Fim), FiU, (syn= FlU), (syn= FiU), F2, (syn= F2), F3, (syn= F3), F4, (syn= F4), F5, (syn= F5), F6, F7, (syn= F7), F25, (syn= F25), (syn= £25), F25U, (syn= F25u), (syn= F25U), (syn= F25V), F44, (syn- F44), F45, (syn= F45), F48, (syn= F48), I, Im, M, MC / 75, M51, (syn= M85), P, (syn= D),S708,R,Tb,(syn= TB),(syn= Tbilisi),W,(syn= Wb),(syn= Weybridge),X,3,6,7,10 / 1,(syn= 10),(syn= F8),(syn= F8),12m,24 / 11,(syn= 24),(syn= F9), (syn= F9), 45 / 111, (syn= 45), 75, 84, 212 / XV, (syn= 212), (syn= Fi0), (syn= FlO), 371 / XXIX, (syn= 371), (syn= Fn), (syn= Fl l) and 513.

[0042] Bacteria of the genus Burkholderia can be infected by the following phages: CP75, NN-Burkholderia (1) and 42.

[0043] Bacteria of the Campylobacter genus can be infected by the following phages: C type, NTCC12669, NTCC12670, NTCC12671, NTCC12672, NTCC12673, NTCC12674, NTCC12675, NTCC12676, NTCC12677, NTCC12678, NTCC12679, NTCC12680, NTCC12681, NTCC12682, NTCC12683, NTCC12684, 32f, 111c, 191, NN-Campylobacter (2), Vfi-6, (syn= V19), VfV-3, V2, V3, V8, V16, (syn= Vfi-1), V19, V20 (V45), V46, (syn= Vfi-2), V5, V6, V7, V8, V9, V10, V11, V12, V13, V14, V15, V16, V17, V18, V19, V20 (V45), V19, V21, V22, V23, V24, V25, V26, V27, V28, V29, V30, V31, V32, V33, V34, V35, V36, V37, V38, V39, V46, V47, V48, V49, V49, V49, V50, V51, V52, V53, V54, V55, V56, V57, V58, V59, V59, V59, V60, V61, V62, V63, V64, V65, V66, V67, V68, V69, V69, V69, V-45) and NN-Campylobacter (1).

[0044] Bacteria of the genus Chlamydia can be infected by the following phages: Chpl.

[0045] Clostridium bacteria can be infected by the following phages: CAKl, CA5, Ca7, CEβ, (syn= 1C), CEγ, Cldl, c-n71, c-203 Tox-, DEβ, (syn= ID), (syn= lDt0X+), HM3, KMl, KT, Ms, NAl, (syn= Naltox+), PA135Oe, Pfo, PL73, PL78, PL81, Pl, P50, P5771, P19402, lCt0X+, 2Ct0X\ 2D3 (syn= 2Dt0X+), 3C, (syn= 3Ctox+), 4C, (syn= 4Ct0X+), 56, III-l, NN-Clostridium (61), NBlt0X+, αl, CAl, HMT, HM2, PFl5 P-23, P-46, Q-05, Q-oe, Q-16, Q-21, Q-26, Q-40, Q-46, S111, SA02, WA01, WA03, Wm, W523, 80, C, CA 2, CA3, CPTl, CPT4, cl, c4, c5, HM7, H11 / A1, H18 / Ax, FWS23, Hi58ZA1, K2ZA1, K21ZS23, ML, NA2t0X; Pf2, Pf3, Pf4, S9ZS3, S41ZA1, S44ZS23, α2, 41, 112ZS23, 214 / S23, 233 / Ai, 234 / S23, 235 / S23, II-l, II-2, II-3, NN-Clostridium (12), CAl, Fl, K, S2, 1, 5, and NN-Clostridium (8).

[0046] Bacteria of the genus Corynebacterium can be infected by the following phages: CGKl (deleted), A, A2, A3, AlOl, A128, A133, A137, A139, A155, A182, B, BF, B17, B18, B51, B271, B275, B276, B277, B279, B282, C, capi, CCl, CGl, CG2, CG33, CL31, Cog, (syn= CG5), D, E, F, H, HI, hqi, hq2, 11ZH33, Ii / 31, J, K, K, (syn= Ktox), L, L, (syn= Ktox) Ltox+), M, MC-I, MC-2, MC-3, MC-4, MLMa, N, O, ovi, ov2, ov3, P, P, R, RP6, RS29, S, T, U, UB1, ub2, UH1, UH3, uh3, uh5, uh6, β, (syn= βtox+), βhv64, βvir, γ, (syn= γtoχ-), γl9, δ, (syn= δ'ox+), p, (syn= ptoχ-), Φ9, φ984, ω, IA, 1 / 1180, 2, 2 / 1180, 5 / 1180, 5ad / 9717, 7 / 4465, 8 / 4465, 8ad / 10269, 10 / 9253, 13Z9253, 15 / 3148, 21 / 9253, 28, 29, 55, 2747, 2893, 4498 and 5848.

[0047] Bacteria of the genus Enterococcus are infected by the following phages: DF78, Fl, F2, 1, 2, 4, 14, 41, 867, Dl, SB24, 2BV, 182, 225, C2, C2F, E3, E62, DS96, H24, M35, P3, P9, SBlOl, S2, 2BII, 5, 182a, 705, 873, 881, 940, 1051, 1057, 21096C, NN-Enterococcus (1), PEl, Fl, F3, F4, VD13, 1, 200, 235, and 341.

[0048] Bacteria of the genus Eysipelothrix can be infected by the following phages: NN-Eysipelothrix (1).

[0049] Bacteria of the genus Escherichia can be infected by the following phages: BW73, B278, D6, D108, E, El, E24, E41, FI-2, FI-4, FI-5, HI8A, Ffl8B, i, MM, Mu, (syn= mu), (syn= MuI), (syn= Mu-I), (syn= MU-I), (syn= MuI), (syn= μ), 025, PhI-5, Pk, PSP3, Pl, PlD, P2, P4(deletion), Sl, Wφ, φK13, φR73(deletion), φl, φ2, φ7, φ92, ψ(deletion), 7 A, 8φ, 9φ, 15 (deletion), 18, 28-1, 186, 299, HH-Escherichia (2), AB48, CM, C4, C16, DD-VI, (syn= Dd-Vi), (syn= DDVI), (syn= DDVi), E4, E7, E28, FIl, FI3, H, Hl, H3, H8, K3, M, N, ND-2, ND-3, ND4, ND-5, ND6, ND-7, Ox-I (syn= OXl), (syn= HF), Ox-2 (syn= 0x2), (syn= 0X2), Ox-3, Ox-4, Ox-5, (syn= 0X5), Ox-6, (syn= 66F), (syn= φ66t), (syn= φ66t-)5 0111, PhI-I, RB42, RB43, RB49, RB69, S, SaI-I, Sal-2, Sal-3, Sal-4, Sal-5, Sal-6, TC23, TC45, TuII * -6, (syn= TuII * ), TuIP-24, TuII *46、TuIP-60、T2、(syn= ganuTia)、(syn= γ)、(syn= PC)、(syn= P.C.)、(syn= T-2)、(syn= T2)、(syn= P4)、T4、(syn= T-4)、(syn= T4)、T6、T35、αl、1、IA、3、(syn= Ac3)、3A、3T+、(syn= 3)、(syn= Ml)、5φ、(syn= φ5)、9266Q、CFO103、HK620、J、K、KlF、m59、no. A、no. E、no. 3、no. 9、N4、sd、(syn= Sd)、(syn= SD)、(syn= Sa)3 (syn= sd)、(syn= SD)、(syn= CD)、T3、(syn= T-3)、(syn= T3)、T7、(syn= T-7)、(syn= T7)、WPK、W31、ΔH、φC3888、φK3、φK7、φK12、φV-1、Φ04-CF、Φ05、Φ06、Φ07、φl、φl.2、φ20、φ95、φ263、φlO92、φl、φll、(syn= φW)、Ω8、1、3、7、8、26、27、28-2、29、30、31、32、38、39、42、933W、NN-エシェリキア(1)、Esc-7-11、AC30、CVX-5、Cl、DDUP、ECl、EC2、E21、E29、Fl、F26S、F27S、Hi、HK022、HK97、(syn= ΦHK97)、HK139、HK253、HK256、K7、ND-I、no.D, PA-2, q, S2, Tl, (syn= α), (syn= P28), (syn= TI), (syn= Tx), T3C, T5, (syn= T-5), (syn= T5), UC-I, w, β4, γ2, λ(syn= lambda), (syn= Φλ), ΦD326, φγ, Φ06, Φ7, Φ10, φ80, χ, (syn= χi), (syn= φχ), (syn= φχi), 2, 4, 4A, 6, 8A, 102, 150, 168, 174, 3000, AC6, AC7, AC28, AC43, AC50, AC57, AC81, AC95, HK243, KlO, ZG / 3A, 5, 5A, 21EL, H19-J, 933H, O157 tie pin Gufarge 1~16, JES-2013, 121Q, 172-1, 1720a-02, ADB-2, AKFV33, av-05, bV_EcoS_AHP42, bV_EcoS_AHP24, bC_EcoS_AHS24, bV_EcoS_AKS96, CBA120. .

[0050] Bacteria of the genus Fusobacterium are infected by the following phages: NN-Fusobacterium (2), fv83-554 / 3, fv88-531 / 2, 227, fv2377, fv2527, and fv8501.

[0051] Bacteria of the genus Haemophilus are infected by the following phages: HP1, S2 and N3.

[0052] Bacteria of the genus Helicobacter are infected by the following phages: HP1 and ^^-Helicobacter (1).

[0053] Klebsiella bacteria are infected by the following phages: AIO-2, KI4B, Kl6B, Kl9, (syn= K19), Kl14, Kl15, Kl21, Kl28, Kl29, KI32, Kl33, Kl35, Kl106B, Kl171B, Kl181B, Kl832B, AIO-I, AO-I, AO-2, AO-3, FC3-10, K, Kl1, (syn= KI1), Kl2, (syn= K12), Kl3, (syn= K13), (syn= Kl 70 / 11), Kl4, (syn= K14), Kl5, (syn= K15), Kl6, (syn= K16), Kl7, (syn= K17), Kl8, (syn= K18), Kl19, (syn= K19), Kl27, (syn= K127), Kl31, (syn= K131), Kl35, Kl171B, II, VI, IX, CI-I, Kl4B, Kl8, Kl11, Kl12, Kl13, Kl16, Kl17, Kl18, Kl20, Kl22, Kl23, Kl24, Kl26, Kl30, Kl34, Kl106B, KIi65B, Kl328B, KLXI, K328, P5046, 11, 380, III, IV, VII, VIII, FC3-11, Kl2B, (syn= K12B), Kl25, (syn= K125), Kl42B, (syn= K142), (syn= K142B), Kl181B, (syn= KIl 81), (syn= K1181B), Kl765 / !, (syn= K1765 / 1), Kl842B, (syn= K1832B), Kl937B, (syn= K1937B), Ll, φ28, 7, 231, 483, 490, 632 and 864 / 100.

[0054] Bacteria of the genus Lepitospira are infected by the following phages: LEl, LE3, LE4, and NN-leptospira (1).

[0055] Listeria bacteria are infected by the following phages: A511, 01761, 4211, 4286, (syn= BO54), A005, A006, A020, A500, A502, A511, Al 18, A620, A640, B012, B021, B024, B025, B035, B051, B053, B054, B055, B056, BlOl, BI lO, B545, B604, B653, C707, D441, HSO47, HlOG, H8 / 73, H19, H21, H43, H46, H107, H108, HI lO, H163 / 84, H312, H340, H387, H391 / 73, H684 / 74, H924A, PSA, U153, φMLUP5, (syn= P35), 00241, 00611, 02971A, 02971C, 5 / 476, 5 / 911, 5 / 939, 5 / 11302, 5 / 11605, 5 / 11704, 184, 575, 633, 699 / 694, 744, 9 00, 1090, 1317, 1444, 1652, 1806, 1807, 1921 / 959, 1921 / 11367, 1921 / 11500, 1921 / 11566, 1921 / 12460, 1921 / 12582, 1 967, 2389, 2425, 2671, 2685, 3274, 3550, 3551, 3552, 4276, 4277, 4292, 4477, 5337, 5348 / 11363, 5348 / 11646, 5348 / 12430, 5348 / 12434, 10072, 11355C, 11711A, 12029, 12981, 13441, 90666, 90816, 93253, 907515, 910716, and NN-Listeria (15).

[0056] Bacteria of the genus Morganella are infected by the following phages:47

[0057] Bacteria of the genus Mycobacterium are infected by the following phages: 13, AGl, ALi, ATCC 11759, A2, B.C3, BG2, BKl, BK5, butyricum, BI, B5, B7, B30, B35, Clark, Cl, C2, DNAIII, DSP1, D4, D29, GS4E, (syn= GS4E), GS7, (syn= GS-7), (syn= GS7), IPa, lacticola, Legendre, Leo, L5, (syn= ΦL-5), MC-I, MC-3, MC-4, minetti, MTPHI l, Mx4, MyF3P / 59a, phlei, (syn= phlei 1), phlei 4, and Polonus. II, rabinovitschi, smegmatis, TM4, TM9, TMlO, TM20, Y7, YlO, φ630, IB, IF, IH, 1 / 1, 67, 106, 1430, Bl, (syn= Bol), B24, D, D29, FK, FS, HP, Polonus I, Roy, Rl, (syn= Rl-Myb), (syn= Ri), 11, 31, 40, 50, 103a, 103b, 128, 3111-D, 3215-D and NN-Mycobacterium (1).

[0058] Bacteria of the genus Neisseria are infected by the following phages: Group I, group II and NP1.

[0059] Bacteria of the genus Nocardia are infected by the following phages: MNP8, NJ-L, NS-8, N5 and TtiN-Nocardia.

[0060] Bacteria of the genus Proteus are infected by the following phages: Pm5, 13vir, 2 / 44, 4 / 545, 6 / 1004, 13 / 807, 20 / 826, 57, 67b, 78, 107 / 69, 121, 9 / 0, 22 / 608, 30 / 680, PmI, Pm3, Pm4, Pm6, Pm7, Pm9, PmIO, PmI l, Pv2, πl, φm, 7 / 549, 9B / 2, 10A / 31, 12 / 55, 14, 15, 16 / 789, 17 / 971, 19A / 653, 23 / 532, 25 / 909, 26 / 219, 27 / 953, 32A / 909, 33 / 971, 34 / 13, 65, 5006M, 7480b, VI, 13 / 3a, Clichy 12, π2600, φχ7, 1 / 1004, 5 / 742, 9, 12, 14, 22, 24 / 860, 2600 / D52, Pm8 and 24 / 2514.

[0061] Bacteria of the genus Providencia are infected by the following phages: PL25, PL26, PL37, 9211 / 9295, 9213 / 921 Ib, 9248, 7 / R49, 7476 / 322, 7478 / 325, 7479, 7480, 9000 / 9402 and 9213 / 921 Ia.

[0062] (syn=) Pf-I)、Pf2、Pf3、PP7、PRRl、7s、im-シュードモナス(1)、AI-I、AI-2、B 17、B89、CB3、Col 2、Col 11、Col 18、Col (syn= PBl)、pfl6、PMN17、PPl、PP8、Psal、PsPl、PsP2、PsP3、PsP4、PsP5、PS3、PS17、PTB80、PX4、PX 7、PYOl、PYO2、PYO5、PYO6、PYO9、PYOlO、PYO13、PYO14、PYO16、PYO18、PYO19、PYO20、PYO29、 PYO32, PYO33, PYO35, PYO36, PYO37, PYO38, PYO39, PYO41, PYO42, PYO45, PYO47, PYO48, PYO64, PYO69, PYO103, PlK, SLPl, SL2, S2, UNL-I, wy, Yai, Ya4, Yan, φBE, φCTX, φC17, φKZ, (syn= ΦKZ)、φ-LT、Φmu78、φNZ、φPLS-1、φST-1、φW-14、φ-2、1 / 72、2 / 79、3、3 / DO、4 / 237、5 / 406、6C、6 / 6660、7、7v、7 / 184、8 / 280、9 / 95、10 / 502、11 / DE、12 / 100、12S、16、21、24、25F、27、31、44、68、71、95、109、188、33 7、352、1214、HN-シュードモナス(23)、A856、B26、CI-I、CI-2、C5、D、gh-1、Fl 16、HF、H90、K5、K6、Kl 04、K109、K166、K267、N4、N5、O6N-25P、PE69、Pf、PPN25、PPN3 5、PPN89、PPN91、PP2、PP3、PP4、PP6、PP7、PP8、PP56、PP87、PPl 14、PP206、PP207、PP306、PP651、Psp231a、Pssy401、Pssy9220、psi、PTB2、PTB20、PTB42、PXl、PX 3,P 7, φPLS743, φS-1, 1, 2, 2, 3, 4, 5, 6, 7, 7, 8, 9, 10, 11, 12, 12B, 13, 14 15, 14, 15, 16, 17, 18, 19, 20, 20, 21, 21, 22, 23, 23, 24, 25, 31, 119x, 145, 147, 170, 267, 284, 308, 525, NN-Current (5) of A7, B3 B33, B39, BI-I, C22, D3, D37, D40, D62, D3112, F7, FlO, g, gd, ge, gξ Hwl2, Jb 19. KFl, L°, OXN-32P, O6N-52P, PCH-I, PC13-1, PC35-1, PH2, PH51 PH93, PH132, PMW, PM13, PM57, PM61, PM62, PM63, PM69, PM105, PMl 13 PM681, PM682, PO4, PPl, PP4, PP5, PP64, PP65, PP66, PP71, PP86, PP88, PP92, PP401, PP711, PP891 Pssy41, Pssy42, Pssy403, Pssy404, Pssy420, Pssy923, PS4, PS-IO, Pz, SDl, SLl, SL3, SL5, SM, φC5, φCl l, φCl l-1, φC13, φC15, φMO, φX, φO4, φl l φ240, 2, 2F, 5, 7m, 11, 13, 13 / 441, 14, 20, 24, 40, 45, 49, 61, 73, 148, 160, 198, 218, 222, 236, 242, 246, 249, 258, 269, 295, 297, 309, 318, 342, 350, 351, 357-1, 400-1, HN- GlOl, M6, M6a, Ll, PB2, Pssyl5, Pssy4210, Pssy4220, PYO12, PYO34, PYO4 9. PYO50, PYO51, PYO52, PYO53, PYO57, PYO59, PYO200, PX2, PX5, SL4, φO3, φO6, and 31214.

[0063] The diagram of the rhinoceros is in the form of NN-Rickettsia.

[0064] Bacteria of the genus Salmonella are infected by the following phages: b, Beccles, CT, d, Dundee, f, FeIs 2, GI, GUI, GVI, GVIII, k, K, i, j, L, 01, (syn = 0 - 1), (syn = O1), (syn = O - I), (syn = 7), 02, 03, P3, P9a, PlO, Sab3, Sab5, SanlS, Sanl7, SI, Taunton, ViI, (syn = ViI), 9, im Salmonella(1), N - I, N - 5, N - IO, N - 17, N - 22, 11, 12, 16 - 19, 20.2, 36, 449C / C178, 966A / C259, a, B.A.O.R., e, G4, GUI, L, LP7, M, MG40, N - 18, PSA68, P4, P9c, P22, (syn = P22), (syn = PLT22), (syn = PLT22), P22al, P22 - 4, P22 - 7, P22 - 11, SNT - I, SNT - 2, SP6, Villi, ViIV, ViV, ViVI, ViVII, Worksop, Sj5, ε34, 1,37, 1(40), (syn = φl

[40] ), 1,422, 2, 2.5, 3b, 4, 5, 6,14(18), 8, 14(6,7), 10, 27, 28B, 30, 31, 32, 33, 34, 36, 37, 39, 1412, SNT - 3, 7 - 11, 40.3, c, C236, C557, C625, C966N, g, GV, G5, Gl 73, h, IRA, Jersey, MB78, P22 - 1, P22 - 3, P22 - 12, Sabl, Sab2, Sab2, Sab4, Sanl, San2, San3, San4, San6, San7, San8, San9, Sanl3, Sanl4, Sanl6, Sanl8, Sanl9, San20, San21, San22, San23, San24, San25, San26, SasLl, SasL2, SasL3, SasL4, SasL5, SlBL, SII, ViII, φl, 1, 2, 3a, 3al, 1010, Ym - Salmonella(1), N - 4, SasL6 and 27.

[0065] Serratia bacteria are infected by the following phages: A2P, PS20, SMB3, SMP, SMP5, SM2, V40, V56, ic, ΦCP-3, ΦCP-6, 3M, 10 / la, 20A, 34CC, 34H, 38T, 345G, 345P, 501B, SMB2, SMP2, BC, BT, CW2, CW3, CW4, CW5, Lt232, L2232, L34, L.228, SLP, SMPA, V .43, σ, φCWl, ΦCP6-1, ΦCP6-2, ΦCP6-5, 3T, 5, 8, 9F, 10 / 1, 2OE, 32 / 6, 34B, 34CT, 34P, 37, 41, 56, 56D, 56P, 6OP, 6 1 / 6, 74 / 6, 76 / 4, 101 / 8900, 226, 227, 228, 229F, 286, 289, 290F, 512, 764a, 2847 / 10, 2847 / 1Oa, L.359 and SMBl.

[0066] You can also use the following expression: Fsa(syn= a)(syn=d=(FSD=2d)2d). W2d). HVIII)、SKγ66、(syn= gamma 66)、(syn= yββ)、(syn= γ66b)、SKm、(syn= SIIIb)5 (syn= UI)、SKw、(syn=SIII)、(S=(syn=IC)、、(Syn=IC IV). SIVA.) (syn= IVA)、(syn= KVI)、(syn= Svi)、(syn= VI)、SKvm、(syn= Svm)、(syn= VIII)、SKVΠIA、(syn= SvmA=)、(syn= SvmA=)、 VIIIA). K29)、FlO、(syn= FSlO)、(syn= K31)、I1、(syn= alpha)、(syn= FSa)、(syn= Kl 8)、(syn= α)、I2、(syn= a)、、、= K19)、、= K19 G35). K2)、(syn= KII)、(syn= Sn)、(syn= II)、(syn= Sm)、(syn= SsIV)、(syn= IV)、SK1Va、(syn= Swab)(sIVa=syn=) IVa)、SKV、(syn= K4)、(syn= KV)、(syn= SV)、(syn= SsV)、(syn= V)、SKx、(syn= K9)、(syn= KX)、(syn= SX)、(X=)、= SsX T35)、(syn= 35-50-R)、STvm、(syn=T8345)、(syn= 8345-SO-SR)、W1、(syn= D8)、(syn= FSD8)、W2a、(syn= D2A)、(syn= FS2a)、DD-2、Sf6、FSi=6、(syn=F F6). 1881-SO-R)、γ66、(syn= gamma 66a)、(syn= Ssγ66)、φ2、BIl、DDVII、(syn= DD7)、FSD2b、(syn= W2B)、=Fsyn2= F2)、FS4、(syn= F4)、(syn= F4)、FS5、(syn= F5)、(syn= F5)、FS9、(syn= F9)、(syn= F9)、FI l、P2-S0-S、(syn=G6)-SO、(syn=G36)、 G36). HXn)、SKI、KI、(syn= S1)、(syn= SsI)、(syn= Svπ)、(syn= SsVII)、(syn= KIX)、(syn= S1x) SsIX)、SKXII、(syn= KXII)、(syn= Sxn)、(syn= SsXII)、STi、STffl、STrv、STVi、STvπ、S70、S206、U2-S0-S、3 210-SO-S、3859-SO-S、4020-SO-S、φ3、φ5、φ7、φ8、ϕ9、φlO、ϕl l、ϕl3、ϕl4、ϕl8、SHm、(syn= Hπi)、SHχi、(syn= HXt) or SKxI、(syn= KXI)、(syn= Sχi)、=X= Ss(XπI)

[0067] A、EW、K、Ph5、Ph9:A、EW、K、Ph5、Ph9 (syn= Sb-I)、S3K、Twort、ΦSK311、φ812、06、40、58、119、130、131、200、1623、STCl、(syn= stcl)、STC2、(syn= stc2)、44AHJD、68、ACl、AC2、A6"C"、A9"C"、b581、CA-I、CA-2、CA-3、CA-4、CA-5、DI l, L39x35, L54a, M42, Nl, N2, N3, N4, N5, N7, N8, NlO, Ni l, N12, N13, N14, N16, Ph6, Phl2, Phl4, UC-18, U4, U15, Sl, S2, S3, S4, S5, X2, Z1, φB5-2, φD, ω, 11, (syn= φl l), (syn= P11-M15)、15、28、28A、29、31、31B、37、42D、(syn= P42D)、44A、48、51、52、52A、(syn= P52A)、52B、53、55、69、71、(syn= P71)、71A、72、75、76、77、79、80、80α、82、82A、83 A、84、85、86、88、88A、89、90、92、95、96、102、107、1 08、111、129-26、130、130A、155、157、157A、165、187 、275、275A、275B、356、456、459、471、471A、489、58 1、676、898、1139、1154A、1259、1314、1380、1405、15 63、2148、2638A、2638B、2638C、2731、2792A、2792B、2818、2835、2848A、3619、5841、12100、AC3、A8、AlO A13, b594n, D, HK2, N9, N15, P52, P87, Sl, S6, Z4, φRE, 3A, 3B, 3C, 6, 7, 16, 21, 42B, 42C, 42E, 44, 47, 47A5 47C、51、54、54x1、70、73、75、78、81、82、88、93、94、1 01、105、110、115、129 / 16、174、594n、1363 / 14、2460 and mS-スタフィロコッカス(1)。

[0068] Bacteria of the genus Streptococcus are infected by the following phages: EJ-I, NN-Streptococcus (1), a, Cl, FL0Ths, H39, Cp-I, Cp-5, Cp-7, Cp-9, Cp-IO, AT298, A5, alO / Jl, alO / J2, alO / J5, alO / J9, A25, BTIl, b6, CAl, c20-l, c20-2, DP-I, Dp-4, DTl, ET42, elO, FA101, FEThs, Fκ, FKKIOI, FKLIO, FKP74, FKH, FLOThs, FyIOl, fl, F10, F20140 / 76, g, GT-234, HB3, (syn= HB-3), HB-623, HB-746, M102, O1205, φO1205, PST, PO, Pl, P2, P3, P5, P6, P8, P9, P9, P12, P13, P14, P49, P50, P5 1, P52, P53, P54, P55, P56, P57, P58, P59, P64, P67, P69, P71, P73, P75, P76, P77, P82, P83, P88, sc, sch, sf, SfIl 1, (syn= SFiI l), (syn= φSFill), (syn=ΦSfil l), (syn= φSfil l), sfil9, (syn= SFil9), (syn= φSFil9), (syn= φSfil9), Sfi21, (syn= SFi21), (syn= φSFi21), (syn= φSfi21), ST0, STX, st2, ST2, ST4, S3, (syn= φS3), s265, φ17, φ42, φ57, φ80, φ81, φ82, φ83, φ84, φ85, φ86, φ87, φ88, φ89, φ90, φ91, φ92, φ93, φ94, φ95, φ96, φ97, φ98, φ99, φlOO, φlOl, φlO2, φ227, φ7201, ωl, ω2, ω3, ω4, ω5, ω6, ω8, ωlO, 1, 6, 9, 1OF, 12 / 12, 14, 1 7SR, 19S, 24, 50 / 33, 50 / 34, 55 / 14, 55 / 15, 70 / 35, 70 / 36, 71 / ST15, 71 / 45, 71 / 46, 74F, 79 / 37, 79 / 38, 80 / J 4, 80 / J9, 80 / ST16, 80 / 15, 80 / 47, 80 / 48, 101, 103 / 39, 103 / 40, 121 / 41, 121 / 42, 123 / 43, 123 / 44, 124 / 44, 337 / ST17 and mStreptococcus ( 34 ).

[0069] Bacteria of the genus Treponema are infected by the following phages: NN-Treponema (1).

[0070] The structure of the solvent is as follows: CTXΦ and fs (syn= si) fs2, Ivpf5, Vfl2, Vf33, VPIΦ, VSK, v6, 493, CP-Tl, ET25, kappa, K13 9. Labol) XN-69P, OXN-86, O6N-21P, PB-I, P147, rp-1, SE3, VA-I (syn= VcA-I), VcA-2, VPl, VP2, VP4, VP7, VP8, VP9, ​​VPlO, VP17, VP18, VP19, X29 (syn= 29). d'Herelle)、t、ΦHAWI-1、ΦHAWI-2、ΦHAWI-3、ΦHAWI-4、ΦHAWI-5、ΦHAWI-6、ΦHAWI-7、 SHOW-8, ΦSHOW-9, ΦSHOW-10, ΦHCl-1, ΦHC1-2, ΦHC1-3, ΦHC1-4, ΦHC2-1,>HC2-2, ΦH C2-3, ΦHC2-4, ΦHC3-1, ΦHC3-2, ΦHC3-3, ΦHD1S-1, ΦHD1S-2, ΦHD2S-1, ΦHD2S-2, ΦHD2 S-3, ΦHD2S-4, ΦHD2S-5, ΦHDO-1, ΦHDO-2, ΦHDO-3, ΦHDO-4, ΦHDO-5, ΦHDO-6, ΦKL-33, Φ KL-34, ΦKL-35, ΦKL-36, ΦKWH-2, ΦKWH-3, ΦKWH-4, ΦMARQ-1, ΦMARQ-2, ΦMARQ-3, ΦMOA T-1, ΦO139, ΦPEL1A-1, ΦPEL1A-2, ΦPEL8A-1, ΦPEL8A-2, ΦPEL8A-3, ΦPEL8C-1, ΦPEL8C -2, ΦPEL13A-1, ΦPEL13B-1, ΦPEL13B-2, ΦPEL13B-3, ΦPEL13B-4, ΦPEL13B-5, ΦPEL1 B-6, ΦPEL13B-7, ΦPEL13B-8, ΦPEL13B-9, ΦPEL13B-10, φVP143, φVP253, Φ16, φl38, 1- II, 5, 13, 14, 16, 24, 32, 493, 6214, 7050, 7227 II (syn= group II) (syn= φ2), V, VIII, ~m-ビリオ(13), KVP20, KVP40, nt-1, O6N-22P, P68, el, e2, e3, e4, e5, FK, G, I K, Nt-6, Nl, N2, N3, N4, N5, O6N-34P, OXN-72P, OXN-85P, OXN-100P, P, Ph-I, PL163 / 10, Q. S, T, φ92, 1-9, 37, 51, 57, 70A-8, 72A-4, 72A-10, 110A-4, 333, 4996, I (syn= group I), III (syn= group III), VI, (syn= A-Saratov), ​​VII, IX, lOA-1, 110A-5, 110A-7, hv-1, OXN-52P, P13, P38, P53, P65, P108, Pill, TPl3 VP3, VP6, VP12, VP13, 70A-3, 70A-4, 70A-10, 72A-1, 108A-3, 109-B1, 110A-2, 149, (syn = φl49), IV, (syn = group IV), NN-Vibrio (22), VP5, VP11, VP15, VP16, αl, α2, α3a, α3b, 353B, and HN-Vibrio (7).

[0071] Bacteria of the genus Yersinia are infected by the following phages: H, HI, H-2, H-3, H-4, Lucas 110, Lucas 303, Lucas 404, YerA3, YerA7, YerA20, YerA41, 3 / M64-76, 5 / G394-76, 6 / C753-76, 8 / C239-76, 9 / F18167, 1701, 1710, PST, 1 / F2852-76, D'Herelle, EV, H, Kotljarova, PTB, R, Y, YerA41, φYerO3-12, 3, 4 / C1324-76, 7 / F783-76, 903, 1 / M6176, and Yer2AT.

[0072] More preferably, the bacteriophage is Salmonella virus SKML39, Shigella virus AG3, Dickeya virus Limestone, Dickeya virus RC2014, Escherichia virus CBA120, Escherichia virus PhaxI, Salmonella virus 38, Salmonella virus Det7, Salmonella virus GG32, Salmonella virus PM10, Salmonella virus SFP10, Salmonella virus SH19, Salmonella virus SJ3, Escherichia virus ECML4, Salmonella virus Marshall, Salmonella virus Irsu Maynard, Salmonella virus SJ2, Salmonella virus STML131, Salmonella virus ViI, Erwinia virus Ea2809, Klebsiella virus 0507KN21, Serratia virus IME250, Serratia virus MAM1, Campylobacter virus CP21, Campylobacter virus CP220, Campylobacter virus CPt10, Campylobacter virus IBB35, Campylobacter virus CP81, Campylobacter virus CP30A, Campylobacter virus CPX, Campylobacter virus Rus NCTC12673, Erwinia virus Ea214, Erwinia virus M7, Escherichia virus AYO145A, Escherichia virus EC6, Escherichia virus HY02, Escherichia virus JH2, Escherichia virus TP1, Escherichia virus VpaE1, Escherichia virus wV8, Salmonella virus FelixO1, Salmonella virus HB2014, Salmonella virus Mushroom, Salmonella virus UAB87, Citrobacter virus Moogle, Citrobacter virus Mordin, Escherichia virus Escherichia virus SUSP1, Escherichia virus SUSP2, Aeromonas virus phiO18P, Haemophilus virus HP1, Haemophilus virus HP2, Pasteurella virus F108, Vibrio virus K139, Vibrio virus Kappa, Burkholderia virus phi52237, Burkholderia virus phiE122, Burkholderia virus phiE202, Escherichia virus 186, Escherichia virus P4, Escherichia virus P2, Escherichia virus Wphi, Mannheimia virus PHL101,Pseudomonas virus phiCTX, Ralstonia virus RSA1, Salmonella virus Fels2, Salmonella virus PsP3, Salmonella virus SopEphi, Yersinia virus L413C, Staphylococcus virus G1, Staphylococcus virus G15, Staphylococcus virus JD7, Staphylococcus virus K, Staphylococcus virus MCE2014, Staphylococcus virus P108, Staphylococcus virus Rodi, Staphylococcus virus S253, Staphylococcus virus S25 -4, Staphylococcal virus SA12, Listeria virus A511, Listeria virus P100, Staphylococcal virus Remus, Staphylococcal virus SA11, Staphylococcal virus Stau2, Bacillus virus Camphawk, Bacillus virus SPO1, Bacillus virus BCP78, Bacillus virus TsarBomba, Staphylococcal virus Twort, Enterococcal virus phiEC24C, Lactobacillus virus Lb338-1, Lactobacillus virus LP65, Enterobacter Virus PG7, Escherichia virus CC31, Klebsiella virus JD18, Klebsiella virus PKO111, Escherichia virus Bp7, Escherichia virus IME08, Escherichia virus JS10, Escherichia virus JS98, Escherichia virus QL01, Escherichia virus VR5, Enterobacter virus Eap3, Klebsiella virus KP15, Klebsiella virus KP27, Klebsiella virus Matisse, Klebsiella virus Miro, Citrobacter virus Merlin, Citrobacter virus Irus Moon, Escherichia virus JSE, Escherichia virus phi1, Escherichia virus RB49, Escherichia virus HX01, Escherichia virus JS09, Escherichia virus RB69, Shigella virus UTAM, Salmonella virus S16, Salmonella virus STML198, Vibrio virus KVP40, Vibrio virus nt1, Vibrio virus ValKK3, Escherichia virus VR7, Escherichia virus VR20, Escherichia virus VR25, Escherichia virus VR26, Shigella virus SP18,Escherichia virus AR1, Escherichia virus C40, Escherichia virus E112, Escherichia virus ECML134, Escherichia virus HY01, Escherichia virus Ime09, Escherichia virus RB3, Escherichia virus RB14, Escherichia virus T4, Shigella virus Pss1, Shigella virus Shfl2, Yersinia virus D1, Yersinia virus PST, Acinetobacter virus 133, Aeromonas virus 65, Aeromonas virus Aeh1, Escherichia virus RB16, Escherichia Virus RB32, Escherichia virus RB43, Pseudomonas virus 42, Cronobacter virus CR3, Cronobacter virus CR8, Cronobacter virus CR9, Cronobacter virus PBES02, Pectobacterium virus phiTE, Cronobacter virus GAP31, Escherichia virus 4MG, Salmonella virus SE1, Salmonella virus SSE121, Escherichia virus FFH2, Escherichia virus FV3, Escherichia virus JES2013, Escherichia virus V5, Brevibacillus virus Abouo, Brevibacillus virus Davies, Bacillus virus Agate, Bacillus virus Bobb, Bacillus virus Bp8pC, Erwinia virus Deimos, Erwinia virus Ea35-70, Erwinia virus RAY, Erwinia virus Simmy50, Erwinia virus Special G, Acinetobacter virus AB1, Acinetobacter virus AB2, Acinetobacter virus AbC62, Acinetobacter virus AP22, Arthrobacter virus ArV1, Arthrobacter virus T rina, Bacillus virus AvesoBmore, Bacillus virus B4, Bacillus virus Bigbertha, Bacillus virus Riley, Bacillus virus Spock, Bacillus virus Troll, Bacillus virus Bastille, Bacillus virus CAM003, Bacillus virus Bc431, Bacillus virus Bcp1, Bacillus virus BCP82, Bacillus virus BM15, Bacillus virus Deepblue, Bacillus virus JBP901, Burkholderia virus Bcep1, Burkholderia virus Bcep43,Burkholderia virus Bcep781, Burkholderia virus BcepNY3, Xanthomonas virus OP2, Burkholderia virus BcepMu, Burkholderia virus phiE255, Aeromonas virus 44RR2, Mycobacterium virus Alice, Mycobacterium virus Bxz1, Mycobacterium virus Dandelion, Mycobacterium virus HyRo, Mycobacterium virus I3, Mycobacterium virus Nappy, Mycobacterium virus Sebata, Clostridium Clostridial virus phiC2, Clostridial virus phiCD27, Clostridial virus phiCD119, Bacillus virus CP51, Bacillus virus JL, Bacillus virus Shanette, Escherichia virus CVM10, Escherichia virus ep3, Erwinia virus Asesino, Erwinia virus EaH2, Pseudomonas virus EL, Halomonas virus HAP1, Vibrio virus VP882, Brevibacillus virus Jimmer, Brevibacillus virus Osiris, Pseudomonas virus Ab 03, Pseudomonas virus KPP10, Pseudomonas virus PAKP3, Sinorhizobium virus M7, Sinorhizobium virus M12, Sinorhizobium virus N3, Erwinia virus Machina, Arthrobacter virus Brent, Arthrobacter virus Jawnski, Arthrobacter virus Martha, Arthrobacter virus Sonny, Edwardsiella virus MSW3, Edwardsiella virus PEi21, Escherichia virus Mu, Shigella virus SfMu, Halobacterium virus phiH, Bacillus virus Grass, Bacillus virus NIT1, Bacillus virus SPG24, Aeromonas virus 43, Escherichia virus P1, Pseudomonas virus CAb1, Pseudomonas virus CAb02, Pseudomonas virus JG004, Pseudomonas virus PAKP1, Pseudomonas virus PAKP4, Pseudomonas virus PaP1, Burkholderia virus BcepF1, Pseudomonas virus 141, Pseudomonas virus Ab28, Pseudomonas virus DL60, Pseudomonas virus DL68,Pseudomonas virus F8, Pseudomonas virus JG024, Pseudomonas virus KPP12, Pseudomonas virus LBL3, Pseudomonas virus LMA2, Pseudomonas virus PB1, Pseudomonas virus SN, Pseudomonas virus PA7, Pseudomonas virus phiKZ, Rhizobium virus RHEph4, Ralstonia virus RSF1, Ralstonia virus RSL2, Ralstonia virus RSL1, Aeromonas virus 25, Aeromonas virus 31, Aeromonas virus Aes12, Aeromonas virus Solanum virus Aes508, Aeromonas virus AS4, Stenotrophomonas virus IME13, Staphylococcus virus IPLAC1C, Staphylococcus virus SEP1, Salmonella virus SPN3US, Bacillus virus 1, Geobacillus virus GBSV1, Yersinia virus R1RT, Yersinia virus TG1, Bacillus virus G, Bacillus virus PBS1, Microcystis virus Ma-LMM01, Vibrio virus MAR, Vibrio virus VHML, Vibrio virus VP585, Bacillus virus BPS 13, Bacillus virus Hakuna, Bacillus virus Megatron, Bacillus virus WPh, Acinetobacter virus AB3, Acinetobacter virus Abp1, Acinetobacter virus Fri1, Acinetobacter virus IME200, Acinetobacter virus PD6A3, Acinetobacter virus PDAB9, Acinetobacter virus phiAB1, Escherichia virus K30, Klebsiella virus K5, Klebsiella virus K11, Klebsiella virus Kp1, Klebsiella virus KP32, Klebsiella virus K30 Klebsiella virus KpV289, Klebsiella virus F19, Klebsiella virus K244, Klebsiella virus Kp2, Klebsiella virus KP34, Klebsiella virus KpV41, Klebsiella virus KpV71, Klebsiella virus KpV475, Klebsiella virus SU503, Klebsiella virus SU552A, Pantoea virus Limelight, Pantoea virus Limezero, Pseudomonas virus LKA1, Pseudomonas virus phiKMV, Xanthomonas virus f20, Xanthomonas virus f30,Xylella virus Prado, Erwinia virus Era103, Escherichia virus K5, Escherichia virus K1-5, Escherichia virus K1E, Salmonella virus SP6, Escherichia virus T7, Kluyvera virus Kvp1, Pseudomonas virus gh1, Prochlorococcus virus PSSP7, Synechococcus virus P60, Synechococcus virus Syn5, Streptococcus virus Cp1, Streptococcus Coccal virus Cp7, Staphylococcal virus 44AHJD, Streptococcal virus C1, Bacillus virus B103, Bacillus virus GA1, Bacillus virus phi29, Kurthia virus 6, Actinomyces virus Av1, Mycoplasma virus P1, Escherichia virus 24B, Escherichia virus 933W, Escherichia virus Min27, Escherichia virus PA28, Escherichia virus Stx2 II, Shigellavirus 7502Stx, Shigellavirus POCJ13, Escherichiavirus 191, Escherichiavirus PA2, Escherichiavirus TL2011, Shigellavirus VASD, Burkholderiavirus Bcep22, Burkholderiavirus Bcepil02, Burkholderiavirus Bcepmigl, Burkholderiavirus DC1, Bordetellavirus BPP1, Burkholderiavirus BcepC6B, Cernophagavirus Cba41, Cernophagavirus Cba172, Dinorceobactervirus DFL12, Erwiniavirus Ea9-2, Erwiniavirus Frozen, Escherichiavirus ph iV10, Salmonella virus Epsilon15, Salmonella virus SPN1S, Pseudomonas virus F116, Pseudomonas virus H66, Escherichia virus APEC5, Escherichia virus APEC7, Escherichia virus Bp4, Escherichia virus EC1UPM, Escherichia virus ECBP1, Escherichia virus G7C, Escherichia virus IME11, Shigella virus Sb1, Achromobacter virus Axp3, Achromobacter virus JWAlpha, Edwardsiella virus KF1, Pseudomonas virus KPP25, Pseudomonas virus R18, Pseudomonas virus Ab09, Pseudomonas virus, Lus LIT1, Pseudomonas virus PA26, Pseudomonas virus Ab22, Pseudomonas virus CHU, Pseudomonas virus LUZ24, Pseudomonas virus PAA2, Pseudomonas virus PaP3, Pseudomonas virus PaP4, Pseudomonas virus TL, Pseudomonas virus KPP21, Pseudomonas virus LUZ7, Escherichia virus N4, Salmonella virus 9NA, Salmonella virus SP069, Salmonella virus BTP1, Salmonella virus HK620, Salmonella virus P22, Salmonella virus ST64T, Shigella virus Sf6, Bacillus virus Page, Bacillus virus Palmer, Bacillus virus Pascal, Bacillus virus Pony, Bacillus virus Pookie, Escherichia virus 172-1, Escherichia virus ECB2, Escherichia virus NJ01, Escherichia virus phiEco32, Escherichia virus Septima11, Escherichia virus SU10, Brucella virus Pr, Brucella virus Tb, Escherichia virus Pollock, Salmonella virus FSL SP-058, Salmonella virus FSL SP-076, Helicobacter virus 1961P, Helicobacter virus KHP30, Helicobacter virus KHP40, Hamiltonella virus APSE1, Lactococcus virus KSY1, Phormidium virus WMP3, Phormidium virus WMP4, Pseudomonas virus 119X, Roseobacter virus SIO1, Vibrio virus VpV262, Vibrio virus VC8, Vibrio virus VP2, Vibrio virus VP5, Streptomyces virus Amela, Streptomyces Streptomyces virus phiCAM, Streptomyces virus Aaronocolus, Streptomyces virus Caliburn, Streptomyces virus Danzina, Streptomyces virus Hydra, Streptomyces virus Izzy, Streptomyces virus Lannister, Streptomyces virus Lika, Streptomyces virus Sujidade, Streptomyces virus Zemlya, Streptomyces virus ELB20, Streptomyces virus R4,Streptomyces virus phiHau3, Mycobacterium virus Acadian, Mycobacterium virus Baee, Mycobacterium virus Reprobate, Mycobacterium virus Adawi, Mycobacterium virus Bane1, Mycobacterium virus BrownCNA, Mycobacterium virus Chrisnmich, Mycobacterium virus Cooper, Mycobacterium virus JAMaL, Mycobacterium virus Nigel, Mycobacterium virus Stinger, My Mycobacterium virus Vincenzo, Mycobacterium virus Zemanar, Mycobacterium virus Apizium, Mycobacterium virus Manad, Mycobacterium virus Oline, Mycobacterium virus Osmaximus, Mycobacterium virus Pg1, Mycobacterium virus Soto, Mycobacterium virus Suffolk, Mycobacterium virus Athena, Mycobacterium virus Bernardo, Mycobacterium virus Gadjet, Mycobacterium Mycobacterium virus Pipefish, Mycobacterium virus Godines, Mycobacterium virus Rosebush, Mycobacterium virus Babsiella, Mycobacterium virus Brujita, Mycobacterium virus Che9c, Mycobacterium virus Sbash, Mycobacterium virus Hawkeye, Mycobacterium virus Plot, Salmonella virus AG11, Salmonella virus Ent1, Salmonella virus f18SE, Salmonella virus Jersey, Salmonella virus L13 , Salmonella virus LSPA1, Salmonella virus SE2, Salmonella virus SETP3, Salmonella virus SETP7, Salmonella virus SETP13, Salmonella virus SP101, Salmonella virus SS3e, Salmonella virus wksl3, Escherichia virus K1G, Escherichia virus K1H, Escherichia virus K1ind1, Escherichia virus K1ind2, Salmonella virus SP31, Leuconostoc virus Lmd1, Leuconostoc virus LN03, Leuconostoc virus LN04,Leuconostoc virus LN12, Leuconostoc virus LN6B, Leuconostoc virus P793, Leuconostoc virus 1A4, Leuconostoc virus Ln8, Leuconostoc virus Ln9, Leuconostoc virus LN25, Leuconostoc virus LN34, Leuconostoc virus LNTR3, Mycobacterium virus Bongo, Mycobacterium virus Rey, Mycobacterium virus Butters, Mycobacterium virus Michelle, Mycobacterium virus Virus Charlie, Mycobacterium virus Pipsqueaks, Mycobacterium virus Xeno, Mycobacterium virus Panchino, Mycobacterium virus Phrann, Mycobacterium virus Redi, Mycobacterium virus Skinnyp, Gordonia virus BaxterFox, Gordonia virus Yeezy, Gordonia virus Kita, Gordonia virus Zirinka, Gordonia virus Nymphadora, Mycobacterium virus Bignus, Mycobacterium virus Brusacoram, Mycobacterium virus Donovan, Mycobacterium virus Fishburne, Mycobacterium virus Jebeks, Mycobacterium virus Malithi, Mycobacterium virus Phayonce, Enterobacter virus F20, Klebsiella virus 1513, Klebsiella virus KLPN1, Klebsiella virus KP36, Klebsiella virus PKP126, Klebsiella virus Sushi, Escherichia virus AHP42, Escherichia virus Rus AHS24, Escherichia virus AKS96, Escherichia virus C119, Escherichia virus E41c, Escherichia virus Eb49, Escherichia virus Jk06, Escherichia virus KP26, Escherichia virus Rogue1, Escherichia virus ACGM12, Escherichia virus Rtp, Escherichia virus ADB2, Escherichia virus JMPW1, Escherichia virus JMPW2, Escherichia virus T1, Shigella virus PSf2, Shigella virus Shfl1, Citrobacter virus Stevie,Escherichia virus TLS, Salmonella virus SP126, Cronobacter virus Esp2949-1, Pseudomonas virus Ab18, Pseudomonas virus Ab19, Pseudomonas virus PaMx11, Arthrobacter virus Amigo, Propionibacterium virus Anatole, Propionibacterium virus B3, Bacillus virus Andromeda, Bacillus virus Blastoid, Bacillus virus Curly, Bacillus virus Eoghan, Bacillus virus Finn, Bacillus virus Glit tering, Bacillus virus Riggi, Bacillus virus Taylor, Gordonia virus Attis, Mycobacterium virus Barnyard, Mycobacterium virus Konstantine, Mycobacterium virus Predator, Mycobacterium virus Bernal13, Staphylococcus virus 13, Staphylococcus virus 77, Staphylococcus virus 108PVL, Mycobacterium virus Bron, Mycobacterium virus Faith1, Mycobacterium virus Joe dirt, Mycobacterium virus Rumpelstiltskin, Lactococcus virus bIL67, Lactococcus virus c2, Lactobacillus virus c5, Lactobacillus virus Ld3, Lactobacillus virus Ld17, Lactobacillus virus Ld25A, Lactobacillus virus LLKu, Lactobacillus virus phiLdb, Cellulophaga virus Cba121, Cellulophaga virus Cba171, Cellulophaga virus Cba181, Cellulophaga virus ST, Bacillus virus 250, Bacillus virus IEBH, Mycobacterium virus Ardmore, Mycobacterium virus Avani, Mycobacterium virus Boomer, Mycobacterium virus Che8, Mycobacterium virus Che9d, Mycobacterium virus Deadp, Mycobacterium virus Dlane, Mycobacterium virus Dorothy, Mycobacterium virus Dotproduct, Mycobacterium virus Drago, Mycobacterium virus Fruitloop, Mycobacterium virus Gumbie,Mycobacterium virus Ibhubesi, Mycobacterium virus Llij, Mycobacterium virus Mozy, Mycobacterium virus Mutaforma13, Mycobacterium virus Pacc40, Mycobacterium virus PMC, Mycobacterium virus Ramsey, Mycobacterium virus Rockyhorror, Mycobacterium virus SG4, Mycobacterium virus Shauna1, Mycobacterium virus Shilan, Mycobacterium virus Spartacus, Mycobacterium Mycobacterium virus Taj, Mycobacterium virus Tweety, Mycobacterium virus Wee, Mycobacterium virus Yoshi, Salmonella virus Chi, Salmonella virus FSLSP030, Salmonella virus FSLSP088, Salmonella virus iEPS5, Salmonella virus SPN19, Mycobacterium virus 244, Mycobacterium virus Bask21, Mycobacterium virus CJW1, Mycobacterium virus Eureka, Mycobacterium virus Kostya, Mycobacterium Umvirus Porky, Mycobacterium virus Pumpkin, Mycobacterium virus Sirduracell, Mycobacterium virus Toto, Mycobacterium virus Corndog, Mycobacterium virus Firecracker, Rhodobacter virus RcCronus, Pseudomonas virus D3112, Pseudomonas virus DMS3, Pseudomonas virus FHA0480, Pseudomonas virus LPB1, Pseudomonas virus MP22, Pseudomonas virus MP29, Pseudomonas virus Rus MP38, Pseudomonas virus PA1KOR, Pseudomonas virus D3, Pseudomonas virus PMG1, Arthrobacter virus Decurro, Gordonia virus Demosthenes, Gordonia virus Katyusha, Gordonia virus Kvothe, Propionibacterium virus B22, Propionibacterium virus Doucette, Propionibacterium virus E6, Propionibacterium virus G4, Burkholderia virus phi6442, Burkholderia virus phi1026b,Burkholderia virus phiE125, Edwardsiella virus eiAU, Mycobacterium virus Ff47, Mycobacterium virus Muddy, Mycobacterium virus Gaia, Mycobacterium virus Giles, Arthrobacter virus Captnmurica, Arthrobacter virus Gordon, Gordonia virus GordTnk2, Paenibacillus virus Harrison, Escherichia virus EK99P1, Escherichia virus HK578, Escherichia virus JL1, Escherichia virus SSL2009a, Escherichia virus YD2008s, Shigella virus EP23, Sodalis virusSO1, Escherichia virus HK022, Escherichia virus HK75, Escherichia virus HK97, Escherichia virus HK106, Escherichia virus HK446, Escherichia virus HK542, Escherichia virus HK544, Escherichia virus HK633, Escherichia virus mEp234, Escherichia virus mEp235, Escherichia virus mEpX1, Escherichia virus mEpX2, Escherichia virus mEp043, Escherichia virus mEp213, Escherichia virus mEp237, Escherichia virus mEp390, Escherichia virus mEp460, Escherichia virus mEp505, Escherichia virus mEp506, Brevibacillus virus Jenst, Achromobacter virus S83-24, Achromobactervirus JWX, Arthrobactervirus Kellezzio, Arthrobactervirus Kitkat, Arthrobactervirus Bennie, Arthrobactervirus DrRobert, Arthrobactervirus Glenn, Arthrobactervirus HunterDalle, Arthrobactervirus Joann, Arthrobactervirus Korra, Arthrobactervirus Preamble, Arthrobactervirus Pumancara, Arthrobactervirus Wayne, Mycobacteriumvirus Alma, Mycobacteriumvirus Arturo, Mycobacteriumvirus Astro, Mycobacteriumvirus Backyardigan, Mycobacteriumvirus BBPiebs31,Mycobacterium virus Benedict, Mycobacterium virus Bethlehem, Mycobacterium virus Billknuckles, Mycobacterium virus Bruns, Mycobacterium virus Bxb1, Myco, Mycobacterium virus Bxz2, Mycobacterium virus Che12, Mycobacterium virus Cuco, Mycobacterium virus D29, Mycobacterium virus Doom, Mycobacterium virus Ericb, Mycobacterium virus Euphoria, Mycobacterium virus George, Mycobacterium virus Gladiator, Mycobacterium virus Goose, Mycobacterium virus Hammer, Mycobacterium virus Heldan, Mycobacterium virus Jasper , Mycobacterium virus JC27, Mycobacterium virus Jeffabunny, Mycobacterium virus JHC117, Mycobacterium virus KBG, Mycobacterium virus Kssjeb, Mycobacterium virus Kugel, Mycobacterium virus L5, Mycobacterium virus Lesedi, Mycobacterium virus LHTSCC, Mycobacterium virus lockley, Mycobacterium virus Marcell, Mycobacterium virus Microwolf, Mycobacterium Virus Mrgordo, Mycobacterium Virus Museum, Mycobacterium Virus Nepal, Mycobacterium Virus Packman, Mycobacterium Virus Peaches, Mycobacterium Virus Perseus, Mycobacterium Virus Pukovnik, Mycobacterium Virus Rebeuca, Mycobacterium Virus Redrock, Mycobacterium Virus Ridgecb, Mycobacterium Virus Rockstar, Mycobacterium Virus Saintus, Mycobacterium Mycobacterium virus Skipole, Mycobacterium virus Solon, Mycobacterium virus Switzer, Mycobacterium virus SWU1, Mycobacterium virus Ta17a, Mycobacterium virus Tiger, Mycobacterium virus Timshel, Mycobacterium virus Trixie, Mycobacterium virus Turbido, Mycobacterium virus Twister, Mycobacterium virus U2, Mycobacterium virus Violet, Mycobacterium virus Wonder,Escherichia virus DE3, Escherichia virus HK629, Escherichia virus HK630, Escherichia virus Lambda, Arthrobacter virus Laroye, Mycobacterium virus Halo, Mycobacterium virus Liefie, Mycobacterium virus Marvin, Mycobacterium virus Mosmoris, Arthrobacter virus Circum, Arthrobacter virus Mudcat, Escherichia virus N15, Escherichia virus 9g, Escherichia virus JenK1, Escherichia virus Virus JenP1, Escherichia virus JenP2, Pseudomonas virus NP1, Pseudomonas virus PaMx25, Mycobacterium virus Baka, Mycobacterium virus Courthouse, Mycobacterium virus Littlee, Mycobacterium virus Omega, Mycobacterium virus Optimus, Mycobacterium virus Thibault, Polaribacter virus P12002L, Polaribacter virus P12002S, Non-Lavens virus P12024L, Non-Lavens virus P 12024S, Thermus virus P23-45, Thermus virus P74-26, Listeria virus LP26, Listeria virus LP37, Listeria virus LP110, Listeria virus LP114, Listeria virus P70, Propionibacterium virus ATCC29399BC, Propionibacterium virus ATCC29399BT, Propionibacterium virus Attacne, Propionibacterium virus Keiki, Propionibacterium virus Kubed, Propionibacterium virus Lauchell y, Propionibacterium virus MrAK, Propionibacterium virus Ouroboros, Propionibacterium virus P91, Propionibacterium virus P105, Propionibacterium virus P144, Propionibacterium virus P1001, Propionibacterium virus P1.1, Propionibacterium virus P100A, Propionibacterium virus P100D, Propionibacterium virus P101A, Propionibacterium virus P104A, Propionibacterium virus PA6,Propionibacterium virus Pacnes201215, Propionibacterium virus PAD20, Propionibacterium virus PAS50, Propionibacterium virus PHL009M11, Propionibacterium virus PHL025M00, Propionibacterium virus PHL037M02, Propionibacterium virus PHL041M10, Propionibacterium virus PHL060L00, Propionibacterium virus PHL067M01, Propionibacterium virus PHL070N0 ...0L00, Propionibacterium virus PHL067M01, Propionibacterium virus PHL070N00, Propionibacterium virus PHL060L00, Propionibacterium virus PHL060L00, Propionibacterium virus PHL067M01, Propionibacterium virus PHL070N00, Propionibacterium virus PHL060L00, Propionibacterium virus PHL060L00, Propionibacterium virus PHL067M01, Propionibacterium virus Pionibacterium virus PHL071N05, Propionibacterium virus PHL082M03, Propionibacterium virus PHL092M00, Propionibacterium virus PHL095N00, Propionibacterium virus PHL111M01, Propionibacterium virus PHL112N00, Propionibacterium virus PHL113M01, Propionibacterium virus PHL114L00, Propionibacterium virus PHL116M00, Propionibacterium virus PHL117M00, Propionibacterium virus PHL117M01, Propionibacterium virus PHL132N00, Propionibacterium virus PHL141N00, Propionibacterium virus PHL151M00, Propionibacterium virus PHL151N00, Propionibacterium virus PHL152M00, Propionibacterium virus PHL163M00, Propionibacterium virus PHL171M01, Propionibacterium virus PHL179M0, Propionibacterium virus PHL194M 00, Propionibacterium virus PHL199M00, Propionibacterium virus PHL301M00, Propionibacterium virus PHL308M00, Propionibacterium virus Pirate, Propionibacterium virus Procrass1, Propionibacterium virus SKKY, Propionibacterium virus Solid, Propionibacterium virus Stormborn, Propionibacterium virus Wizzo, Pseudomonas virus PaMx28, Pseudomonas virus PaMx74,Mycobacterium virus Patience, Mycobacterium virus PBI1, Rhodococcus virus Pepy6, Rhodococcus virus Poco6, Propionibacterium virus PFR1, Streptomyces virus phiBT1, Streptomyces virus phiC31, Streptomyces virus TG1, Caulobacter virus Karma, Caulobacter virus Magneto, Caulobacter virus phiCbK, Caulobacter virus Rogue, Caulobacter virus Swift, Staphylococcus virus Staphylococcal virus 11, Staphylococcal virus 29, Staphylococcal virus 37, Staphylococcal virus 53, Staphylococcal virus 55, Staphylococcal virus 69, Staphylococcal virus 71, Staphylococcal virus 80, Staphylococcal virus 85, Staphylococcal virus 88, Staphylococcal virus 92, Staphylococcal virus 96, Staphylococcal virus 187, Staphylococcal virus 52a, Staphylococcal virus 80alpha, Staphylococcal virus CNPH82, Staphylococcal virus EW, Staphylococcal virus IPLA5, Staphylococcal virus IPLA7, Staphylococcal virus IPLA88, Staphylococcal virus PH15, Staphylococcal virus phiETA, Staphylococcal virus phiETA2, Staphylococcal virus phiETA3, Staphylococcal virus phiMR11, Staphylococcal virus phiMR25, Staphylococcal virus phiNM1, Staphylococcal virus phiNM2, Staphylococcal Lactobacillus virus phiNM4, Staphylococcus virus SAP26, Staphylococcus virus X2, Enterococcus virus FL1, Enterococcus virus FL2, Enterococcus virus FL3, Lactobacillus virus ATCC8014, Lactobacillus virus phiJL1, Pediococcus virus cIP1, Aeromonas virus pIS4A, Listeria virus LP302, Listeria virus PSA, Methanobacterium virus psiM1, Roseobacter virus RDJL1, Roseobacter virus RDJL2,Rhodococcus virus RER2, Enterococcus virus BC611, Enterococcus virus IMEEF1, Enterococcus virus SAP6, Enterococcus virus VD13, Streptococcus virus SPQS1, Mycobacterium virus Papyrus, Mycobacterium virus Send513, Burkholderia virus KL1, Pseudomonas virus 73, Pseudomonas virus Ab26, Pseudomonas virus Kakheti25, Escherichia virus Cajan, Escherichia virus Seurat, Staphylococcus aureus Staphylococcus virus SEP9, Staphylococcus virus Sextaec, Streptococcus virus 858, Streptococcus virus 2972, Streptococcus virus ALQ132, Streptococcus virus O1205, Streptococcus virus Sfi11, Streptococcus virus 7201, Streptococcus virus DT1, Streptococcus virus phiAbc2, Streptococcus virus Sfi19, Streptococcus virus Sfi21, Paenibacillus virus Diva, Paenibacillus virus Hb10c2, Paenibacillus virus Rani, Paenibacillus virus Shelly, Paenibacillus virus Sitara, Paenibacillus virus Willow, Lactococcus virus 712, Lactococcus virus ASCC191, Lactococcus virus ASCC273, Lactococcus virus ASCC281, Lactococcus virus ASCC465, Lactococcus virus ASCC532, Lactococcus virus Bibb29, Lactococcus virus bIL170, Lactococcus virus CB13, Lactococcus virus CB14, Lactococcus virus CB19, Lactococcus virus CB20, Lactococcus virus jj50, Lactococcus virus P2, Lactococcus virus P008, Lactococcus virus sk1, Lactococcus virus Sl4, Bacillus virus Slash, Bacillus virus Stahl, Bacillus virus Staley, Bacillus virus Stills, Gordonia virus Bachita, Gordonia virus ClubL, Gordonia virus OneUp, Gordonia virus Smoothie, Gordonia virus Soups,Bacillus virus SPbeta, Vibriovirus MAR10, Vibriovirus SSP002, Escherichia virus AKFV33, Escherichia virus BF23, Escherichia virus DT57C, Escherichia virus EPS7, Escherichia virus FFH1, Escherichia virus H8, Escherichia virus slur09, Escherichia virus T5, Salmonella virus 118970sal2, Salmonella virus Shivani, Salmonella virus SPC35, Salmonella virus Stitch, Arthrobacter virus Tan k, Tsukamurella virus TIN2, Tsukamurella virus TIN3, Tsukamurella virus TIN4, Rhodobacter virus RcSpartan, Rhodobacter virus RcTitan, Mycobacterium virus Anaya, Mycobacterium virus Angelica, Mycobacterium virus Crimd, Mycobacterium virus Fionnbarth, Mycobacterium virus Jaws, Mycobacterium virus Larva, Mycobacterium virus Macncheese, Mycobacterium virus Pixie, My Cobacterium virus TM4, Bacillus virus BMBtp2, Bacillus virus TP21, Geobacillus virus Tp84, Staphylococcus virus 47, Staphylococcus virus 3a, Staphylococcus virus 42e, Staphylococcus virus IPLA35, Staphylococcus virus phi12, Staphylococcus virus phiSLT, Mycobacterium virus 32HC, Rhodococcus virus RGL3, Paenibacillus virus Vegas, Gordonia virus Vendetta, Bacillus virus Wbeta, Mycobacterial virus Wildcat, Gordonia virus Twister6, Gordonia virus Wizard, Gordonia virus Hotorobo, Gordonia virus Monty, Gordonia virus Woes, Xanthomonas virus CP1, Xanthomonas virus OP1, Xanthomonas virus phil7, Xanthomonas virus Xop411, Xanthomonas virus Xp10, Streptomyces virus TP1604, Streptomyces virus YDN12, Alphaproteobacterial virus phiJl001,Pseudomonas virus LKO4, Pseudomonas virus M6, Pseudomonas virus MP14, 12, Pseudomonas virus PAE1, Pseudomonas virus Yua, Pseudoalteromonas virus PM2, Pseudomonas virus phi6, Pseudomonas virus phi8, Pseudomonas virus phi12, Pseudomonas virus phi13, Pseudomonas virus phi2954, Pseudomonas virus phiNN, Pseudomonas virus phiYY, Vibrio virus fs1, Vibrio virus VGJ, Ralstonia virus RS603, Ralstonia virus RSM1, Ralstonia virus RSM3, Escherichia coli Avirus M13, Escherichia virus I22, Salmonella virus IKe, Acholeplasma virus L51, Vibrio virus fs2, Vibrio virus VFJ, Escherichia virus If1, Propionibacterium virus B5, Pseudomonas virus Pf1, Pseudomonas virus Pf3, Ralstonia virus PE226, Ralstonia virus RSS1, Spiroplasma virus SVTS2, Stenotrophomonas virus PSH1, Stenotrophomonas virus SMA6, Stenotrophomonas virus SMA7, Stenotrophomonas virus Vibriovirus SMA9, Vibriovirus CTXphi, Vibriovirus KSF1, Vibriovirus VCY, Vibriovirus Vf33, Vibriovirus VfO3K6, Xanthomonas virus Cf1c, Spiroplasma virus C74, Spiroplasma virus R8A2B, Spiroplasma virus SkV1CR23x, Escherichia virus FI, Escherichia virus Qbeta, Escherichia virus BZ13, Escherichia virus MS2, Escherichia virus alpha3, Escherichia virus ID21, Escherichia virus ID32, Escherichia virus ID62, Escherichia virus NC28, Escherichia virus NC29, Escherichia virus NC35, Escherichia virus phiK, Escherichia virus St1, Escherichia virus WA45, Escherichia virus G4, Escherichia virus ID52, Escherichia virus Talmos, Escherichia virus phiX174, Bdellovibriovirus MAC1, Bdellovibriovirus MH2K, Chlamydia virus Chp1, Chlamydia virus Chp2, Chlamydia virus CPAR39,Chlamydia virus CPG1, Spiroplasma virus SpV4, Acholeplasma virus L2, Pseudomonas virus PR4, Pseudomonas virus PRD1, Bacillus virus AP50, Bacillus virus Bam35, Bacillus virus GIL16, Bacillus virus Wip1, Escherichia virus phi80, Escherichia virus RB42, Escherichia virus T2, Escherichia virus T3, Escherichia virus T6, Escherichia virus VT2-Sa, Escherichia virus VT1-Sakai, Escherichia virus VT2-Sakai, Escherichia virus CP-933V, Escherichia virus P27, Escherichia virus Stx2phi-I, Escherichia virus Stx1phi, Selected from the group consisting of Escherichia virus Stx2phi-II, Escherichia virus CP-1639, and based on Escherichia virus BP-4795, Escherichia virus 86, Escherichia virus Min27, Escherichia virus 2851, Escherichia virus 1717, Escherichia virus YYZ-2008, Escherichia virus EC026_P06, Escherichia virus ECO103_P15, Escherichia virus ECO103_P12, Escherichia virus ECO111_P16, Escherichia virus ECO111_P11, Escherichia virus VT2phi_272, Escherichia virus TL-2011c, Escherichia virus P13374, Escherichia virus Sp5,

[0073] Additionally, it is a manufacturer's license plate for the BW73.B 278, D6, D108, E, El, E24, E41, FI-2, FI-4, FI- 5, HI8A, Ffl8B, i, MM, Mu, 025, PhI-5, Pk, PSP3 Pl, PlD, P2, P4, Sl, Wφ, φK13, φl, φ2, φ7, φ92, A, 8φ, 9φ, 18, 28-1, 186, 299, HH-Carlich (2), AB48, CM, C4, C16, DD-VI E4, E7, E28, FIl, FI3, H, Hl, H3, H8, K3, M, N, ND-2, ND-3, ND4, ND-5, ND 6, ND-7, Ox-I, Ox-2, Ox-3, Ox-4, Ox-5, Ox-6, PhI-I, RB42, RB43, RB49 RB69, S, SaI-I, Sal-2, Sal-3, Sal-4, Sal-5, Sal-6, TC23, TC45, TuII * -6, TuIP-24, TuII*46, TuIP-60, T2, T4, T6, T35, αl, 1, IA 3A, 3T+, 5φ, 9266Q, CFO103, HK620, J, K, KlF, m59, no. A、no. Yes, no. 3、no. 9, N4, sd, T3, T7, WPK, W31, ΔH, φC3888, φK3, φK7, φK12, φV-1, Φ04-CF, Φ 05, Φ06, Φ07, φl, φl.2, φ20, φ95, φ263, φlO92, φl, φll, Ω8, 1, 3, 7, 8, 27. 28-2. 29. 30. 31. 32. 38. 39. 42. 933W C30, CVX-5, Cl, DDUP, ECl, EC2, E21, E29, Fl, F26S, F27S, Hi, HK022, HK 97, HK139, HK253, HK256, K7, ND-I, PA-2, q, S2, Tl,) T3C, T5, UC-I, w. β4, γ2, λ, ΦD326, φγ, Φ06, Φ7, Φ10, φ80, χ, 2, 4, 4A, 6, 8A, 102, 150, 168; 174, 3000, AC6, AC7, AC28, AC43, AC50, AC57, AC81, AC95, HK243, KlO, Z G / 3A, 5, 5A, 21EL, H19-J and 933H It is a great way to get your hands on.

[0074] origin of replication Replication origins known in the art have been identified from species-specific plasmid DNA (e.g., CoIE1, R1, pT181, pSC101, pMB1, R6K, RK2, p15a, and the like), bacterial viruses (e.g., φX174, M13, F1, and P4), and bacterial chromosomal replication origins (e.g., oriC).

[0075] In one embodiment, the nucleic acid payload used in the present invention, preferably a plasmid, comprises a bacterial origin of replication that is functional in the target bacterium.

[0076] Alternatively, the nucleic acid payload used in the present invention, preferably a plasmid, does not contain any functional bacterial origin of replication or contains an origin of replication that is inactive in the target bacterium, and therefore is unable to replicate itself after being introduced into the bacterium by a bacterial viral particle.

[0077] In one embodiment, the origin of replication in the packaged nucleic acid payload, preferably a plasmid, is inactive in the target bacterium, meaning that this origin of replication is not functional in the target bacterium by the bacterial viral particle, thus preventing replication of the unwanted payload.

[0078] In one embodiment, the nucleic acid payload, preferably a plasmid, comprises a bacterial origin of replication that is functional in the bacteria used for production of bacterial viral particles.

[0079] Bacteria-specific origin of replication

[0080] Plasmid replication depends on host enzymes and plasmid-controlled cis- and trans-determinants. For example, some plasmids contain determinants that are recognized by almost all Gram-negative bacteria and function correctly in each host during replication initiation and regulation. Other plasmids possess this ability only in some types of bacteria (Kues, U and Stahl, U, 1989, Microbiol Rev 53:491-516).

[0081] Plasmids replicate by three general mechanisms: theta-type, strand displacement, and rolling circle (reviewed by Del Solar et al., 1998, Microhio and Molec Biol. Rev 62:434-464), which initiate at the origin of replication, which contains sites necessary for the interaction of plasmid- and / or host-encoded proteins.

[0082] The origin of replication used in the nucleic acid payloads, preferably plasmids, used in the present invention can be of medium copy number, such as the ColE1 ori from pBR322 (15-20 copies per cell) or the R6K plasmid, or can be of high copy number, such as the pUC ori (500-700 copies per cell), pGEM ori (300-400 copies per cell), pTZ ori (>1000 copies per cell), or pBluescript ori (300-500 copies per cell).

[0083] In one embodiment, the bacterial origin of replication is ColE1, pMB1, and variants (such as pBR322, pET, pUC), p15a, ColA, ColE2, pOSAK, pSC101, R6K, IncW (such as pSa), IncFII, pT181, P1, FIncP, IncC, IncJ, IncN, IncP1, IncP4, IncQ, IncH11, RSF1010, CloDF13, NTP16, R1, f5, pPS10, pC194, pE194, BBR1, pBC1, pEP2, pWVO1, pLF1311, pAP1 , pWKS1, pLS1, pLS11, pUB6060, pJD4, pIJ101, pSN22, pAMbeta1, pIP501, pIP407, ZM6100(Sa), pCU1, RA3, pMOL98, RK2 / RP4 / RP1 / R68, pB10, R300B, pRO1614, pRO1600, pECB2, pCM1, pFA3, RepFIA, RepFIB, RepFIC, pYVE439-80, R387, phasyl, RA1, TF-FC2, pMV158, and pUB113.

[0084] More preferably, the bacterial origin of replication is an E. coli origin of replication selected from the group consisting of ColE1, pMB1, and variants (pBR322, pET, pUC, etc.), p15a, ColA, ColE2, pOSAK, pSC101, R6K, IncW (pSa, etc.), IncFII, pT181, P1, FIncP, IncC, IncJ, IncN, IncP1, IncP4, IncQ, IncH11, RSF1010, CloDF13, NTP16, R1, f5, and pPS10.

[0085] More preferably, the bacterial origin of replication is selected in the group consisting of pC194, pE194, BBR1, pBC1, pEP2, pWVO1, pLF1311, pAP1, pWKS1, pLS1, pLS11, pUB6060, pJD4, pIJ101, pSN22, pAMbeta1, pIP501, pIP407, ZM6100(Sa), pCU1, RA3, pMOL98, RK2 / RP4 / RP1 / R68, pB10, R300B, pRO1614, pRO1600, pECB2, pCM1, pFA3, RepFIA, RepFIB, RepFIC, pYVE439-80, R387, phasyl, RA1, TF-FC2, pMV158, and pUB113.

[0086] Even more preferably, the bacterial origins of replication are ColE1 and p15a.

[0087] In one embodiment, the bacterial origin of replication is functional in Propionibacterium and Cutibacterium, more particularly in Propionibacterium freudenreichii and Cutibacterium acnes, and is selected from the group consisting of pLME108, pLME106, p545, pRGO1, pZGX01, pPG01, pYS1, FRJS12-3, FRJS25-1, pIMPLE-HL096PA1, A_15_1_R1.

[0088] Phage replication origin A payload according to the present invention may comprise a phage origin of replication capable of initiating replication of the payload by complementation of the complete phage genome for subsequent inclusion into a different capsid.

[0089] Phage origins of replication can also be engineered to function as bacterial origins of replication without the need to package any phage particles.

[0090] The phage origin of replication included in the payload of the present invention can be any origin of replication found in a phage.

[0091] Preferably, the phage origin of replication can be the wild-type or non-wild-type sequence of M13, f1, φX174, P4, lambda, P2, 186, lambda-like, HK022, mEP237, HK97, HK629, HK630, mEP043, mEP213, mEP234, mEP390, mEP460, mEPx1, mEPx2, phi80, mEP234, T2, T4, T5, T7, RB49, phiX174, R17, PRD1P1-like, P2-like, P22, P22-like, N15, and N15-like bacteriophages.

[0092] More preferably, the phage origin of replication is selected in the group consisting of M13, f1, φX174, P4, and lambda phage origins of replication.

[0093] In a particular embodiment, the phage origin of replication is a P4 origin of replication.

[0094] In certain embodiments, the phage origin of replication is derived from a Propionibacterium phage: a BW-like phage, e.g., Dowsett, B22, E6, G4; a BV-like phage, e.g., Anatol, E1, B3; a BX-like phage, e.g., PFR1 and PFR2; a filamentous B5 phage; a BU-like phage (Cutibacterium acnes phage).

[0095] The bacteria targeted by the bacteriophage-derived particles can be any bacteria present in a mammalian organism, in a plant, or in the environment, and can be any commensal, symbiotic, or pathogenic bacterium of the microflora or microbiome.

[0096] The microbiome can include a variety of endogenous cell types, any of which can be targeted by the present disclosure. In some embodiments, the genus and / or species of the targeted endogenous bacterial cells can depend on the type of bacteriophage used to prepare the bacterial viral particles. For example, some bacteriophages exhibit tropism or preferentially target specific host species of bacteria. Other bacteriophages do not exhibit such tropism and can be used to target several different genera and / or species of endogenous bacterial cells.

[0097] Examples of bacterial cells include, but are not limited to, cells from the following genera: Yersinia spp., Escherichia spp., Klebsiella spp., Acinetobacter spp., Bordetella spp., Neisseria spp., Aeromonas spp., Francisella spp., Corynebacterium spp., Citrobacter spp., Chlamydia spp., Haemophilus spp., Brucella spp., and Mycobacterium spp. spp.), Legionella spp., Rhodococcus spp., Pseudomonas spp., Helicobacter spp., Vibrio spp., Bacillus spp., Erysipelothrix spp., Salmonella spp., Streptomyces spp., Streptococcus spp., Staphylococcus spp., Bacteroides spp., Prevotella spp., Clostridium spp. spp.), Bifidobacterium spp., Clostridium spp., Brevibacterium spp., Lactococcus spp., Leuconostoc spp., Actinobacillus spp., Selnomonas spp., Shigella spp., Zymomonas spp.), Mycoplasma spp., Treponema spp., Leuconostoc spp., Corynebacterium spp., Enterococcus spp., Enterobacter spp., Pyrococcus spp., Serratia spp., Morganella spp., Parvimonas spp., Fusobacterium spp., Actinomyces spp., Porphyromonas spp., Micrococcus spp. spp.), Bartonella spp., Borrelia spp., Brucelia spp., Campylobacter spp., Chlamydophilia spp., Cutibacterium spp., Propionibacterium spp., Gardnerella spp., Ehrlichia spp., Haemophilus spp., Leptospira spp., Listeria spp., Mycoplasma spp., Nocardia spp. Examples of suitable bacteria include cells derived from bacteria of the genera Rickettsia, Ureaplasma, Lactobacillus, Faecalibacterium, Ruminococcus, and mixtures thereof.

[0098] Thus, bacteriophage-derived particles may target (e.g., specifically target) bacterial cells from any one or more of the aforementioned genera of bacteria for specific delivery of a payload according to the present invention.

[0099] Preferably, the target bacteria can be selected from the group consisting of Yersinia, Escherichia, Klebsiella, Acinetobacter, Pseudomonas, Helicobacter, Vibrio, Salmonella, Streptococcus, Staphylococcus, Bacteroides, Clostridium, Shigella, Enterococcus, Enterobacter, Listeria, Cutibacterium, Propionibacterium, Fusobacterium, Porphyromonas, and Gardnerella.

[0100] In some embodiments, the bacterial cells of the present invention are anaerobic bacterial cells (e.g., cells that do not require oxygen for growth). Anaerobic bacterial cells include facultative anaerobic cells, such as, but not limited to, Escherichia coli, Shewanella oneidensis, Gardnerella vaginalis, and Listeria. Anaerobic bacterial cells also include obligate anaerobic cells, such as Bacteroides, Clostridium, Cutibacterium, Propionibacterium, Fusobacterium, and Porphyromonas species. In humans, anaerobic bacteria are most commonly found in the gastrointestinal tract. Thus, in some specific embodiments, the target bacteria are bacteria most commonly found in the gastrointestinal tract. The bacteriophages used to prepare the bacterial viral particles, and the bacterial viral particles, can target (e.g., specifically target) anaerobic bacterial cells according to their specific spectrum known to those skilled in the art for specific delivery of plasmids.

[0101] In some embodiments, the target bacterial cells are selected from the group consisting of Bacteroides thetaiotaomicron, Bacteroides fragilis, Bacteroides distasonis, Bacteroides vulgatus, Clostridium leptum, Clostridium coccoides, Staphylococcus aureus, Bacillus subtilis, Clostridium butyricum, Brevibacterium lactofermentum, Streptococcus agalactiae, Lactococcus lactis, and the like. lactis, Leuconostoc lactis, Actinobacillus actinobycetemcomitans, Cyanobacteria, Escherichia coli, Helicobacter pylori, Selenomonas ruminatium, Shigella sonnei, Zymomonas mobilis, Mycoplasma mycoides, Treponema denticola, Bacillus thuringiensis, Staphylococcus lugdunensis lugdunensis, Leuconostoc oenos, Corynebacterium xerosis, Lactobacillus plantarumplantarum, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus acidophilus, Enterococcus faecalis, Bacillus coagulans, Bacillus cereus, Bacillus popillae, Synechocystis strain PCC6803, Bacillus liquefaciens, Pyrococcus abyssi, Selenomonas nominantium nominantium, Lactobacillus hilgardii, Streptococcus ferus, Lactobacillus pentosus, Bacteroides fragilis, Staphylococcus epidermidis, Streptomyces phaechromogenes, Streptomyces ghanaenis, Klebsiella pneumoniae, Enterobacter cloacae, Enterobacter aerogenes, Serratia marcescens marcescens, Morganella morganii, Citrobacter freundii, Propionibacterium freudenreichii, Pseudomonas aerigunosa, Parvimonas micramicra, Prevotella intermedia, Fusobacterium nucleatum, Prevotella nigrescens, Actinomyces israelii, Porphyromonas endodontalis, Porphyromonas gingivalis, Micrococcus luteus, Bacillus megaterium, Aeromonas hydrophila, Aeromonas caviae, Bacillus anthracis, Bartonella henselae henselae, Bartonella Quintana, Bordetella pertussis, Borrelia burgdorferi, Borrelia garinii, Borrelia afzelii, Borrelia recurrentis, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Campylobacter jejuni, Campylobacter coli, Campylobacter fetus, Chlamydia pneumoniae pneumoniae, Chlamydia trachomatis, Chlamydophila psittaci, Clostridium botulinumbotulinum, Clostridium difficile, Clostridium perfringens, Clostridium tetani, Corynebacterium diphtheria, Cutibacterium acnes (formerly Propionibacterium acnes), Ehrlichia canis, Ehrlichia chaffeensis, Enterococcus faecium, Francisella tularensis, Haemophilus influenzae, Legionella pneumophila pneumophila, Leptospira interrogans, Leptospira santarosai, Leptospira weilii, Leptospira noguchii, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium ulcerans, Mycoplasma pneumonia, Neisseria gonorrhoeae, Neisseria meningitides, Nocardia asteroids), Rickettsia rickettsia, Salmonella enteritidis, Salmonella typhi, Salmonella paratyphiparatyphi, Salmonella typhimurium, Shigella flexnerii, Shigella dysenteriae, Staphylococcus saprophyticus, Streptococcus pneumoniae, Streptococcus pyogenes, Gardnerella vaginalis, Streptococcus viridans, Treponema pallidum, Ureaplasma urealyticum, Vibrio cholera, Vibrio parahaemolyticus, Yersinia pestis pestis, Yersinia enterocolitic, Yersinia pseudotuberculosis, Actinobacter baumannii, Pseudomonas aerigunosa, and mixtures thereof, and preferably, the bacterium of interest is selected from the group consisting of Escherichia coli, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter cloacae, and Enterobacter aerogenes, and mixtures thereof.

[0102] In some embodiments, the target bacterial cells are from the genera Anaerotruncus, Acetanaerobacterium, Acetitomaculum, Acetivibrio, Anaerococcus, Anaerofilum, Anaerosinus, Anaerostipes, Anaerovorax, Butyrivibrio, and the like. Butyrivibrio, Clostridium, Capracoccus, Dehalobacter, Dialister, Dorea, Enterococcus, Ethanoligenens, Faecalibacterium, Fusobacterium, Gracilibacter, Gugenhei Guggenheimella, Hespelia, Lachnobacteriium, Lachnospira, Lactobacillus, Leuconostoc, Megamonas, Moriella, Mitsuokella, Oribacterium, Oxobacter, Papillobacter ibacter, Proprionispira, Pseudobutyrivibrio, Pseudoramibacter, Roseburia, Ruminococcus, Sarcina, Seinonella, Shuttleworthia, Sporobacter, Sporobacterium,Streptococcus, Subdoligranulum, Syntrophococcus, Thermobacillus, Turibacter, Weisella, Clostridium, Bacteroides, Ruminococcus, Faecalibacterium erium, Treponema, Phascolarctobacterium, Megasphaera, Faecalibacterium, Bifidobacterium, Lactobacillus, Sutterella, and Prevotella.

[0103] In other embodiments, the target bacterial cells are selected from the group consisting of Achromobacter xylosoxidans, Acidaminococcus fermentans, Acidaminococcus intestini, Acidaminococcus spp., Acinetobacter baumannii, Acinetobacter junii, Acinetobacter lwoffii, Actinobacillus capsulatus, Actinomyces naeslundii, Actinomyces nouii, and the like. neuii, Actinomyces odontolyticus, Actinomyces radingae, Adlercreutzia equolifaciens, Aeromicrobium massiliense, Aggregatibacter actinomycetemcomitans, Akkermansia muciniphila, Aliagarivorans marinus, Alistipes finegoldii, Alistipes indistinctus, Alistipes inops, Alistipes onderdonchii onderdonkii, Alistipes putredinis, Alistipes senegalensis, Alistipes shahii, Alistipes timonensistimonensis, Alloscardovia omnicolens, Anaerobacter polyendosporus, Anaerobaculum hydrogeniformans, Anaerococcus hydrogenalis, Anaerococcus prevotii, Anaerococcus senegalensis, Anaerofustis stercorihominis, Anaerostipes caccae, Anaerostipes hadrus, Anaeroturuncus corihominis colihominis, Aneurinibacillus aneurinilyticus, Bacillus licheniformis, Bacillus massilioanorexius, Bacillus massiliosenegalensis, Bacillus simplex, Bacillus smithii, Bacillus subtilis, Bacillus thuringiensis, Bacillus timonensis, Bacteroides xylanisolvens, Bacteroides acidifaciens acidifaciens, Bacteroides caccae, Bacteroides capillosus, Bacteroides cellulosiliticuscellulosilyticus, Bacteroides clarus, Bacteroides coprocola, Bacteroides coprophilus, Bacteroides dorei, Bacteroides eggerthii, Bacteroides faecis, Bacteroides finegoldii, Bacteroides fluxus, Bacteroides fragilis, Bacteroides gallinarum, Bacteroides intestinalis, Bacteroides nordii nordii, Bacteroides oleiciplenus, Bacteroides ovatus, Bacteroides pectinophilus, Bacteroides plebeius, Bacteroides salanitronis, Bacteroides salyersiae, Bacteroides genus, Bacteroides, Bacteroides stercoris, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides vulgatus, Bacteroides xylanisorbens xylanisolvens, Bacteroides pectinophilus ATCC, Barnesiella intestinihominisintestinihominis, Bavariicoccus seileri, Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium dentium, Bifidobacterium gallicum, Bifidobacterium longum longum, Bifidobacterium pseudocatenulatum, Bifidobacterium stercoris, Bilophila wadsworthia, Blautia faecis, Blautia hansenii, Blautia hydrogenotrophica, Blautia luti, Blautia obeum, Blautia producta, Blautia wexlerae, Brachymonas chironomi, Brevibacterium senegalensis senegalense, Bryantella formatexigens, Clostridium butyricum (butyrate-producingbacterium, Butyricicoccus pullicaecorum, Butyricimonas virosa, Butyrivibrio crossotus, Butyrivibrio fibrisolvens, Caldicoprobacter faecalis, Campylobacter concisus, Campylobacter jejuni, Campylobacter upsaliensis, Catenibacterium mitsuokai, Cedecea davisae davisae, Cellulomonas massiliensis, Cetobacterium somerae, Citrobacter braakii, Citrobacter freundii, Citrobacter pasteurii, Citrobacter spp., Citrobacter youngae, Cloacibacillus evryensis, Clostridiales bacterium, Clostridioides difficile, Clostridium asparagiforme, Clostridium bartlettii bartlettii, Clostridium boliviensis, Clostridium bolteae, Clostridium hasewaiihathewayi, Clostridium hiranoni, Clostridium hylemonae, Clostridium leptum, Clostridium methylpentosum, Clostridium nexile, Clostridium orbiscindens, Clostridium ramosum, Clostridium scindens, Clostridium spp., Clostridium spiroforme, Clostridium sporogenes, Clostridium symbiosum symbiosum, Collinsella aerofaciens, Collinsella intestinalis, Collinsella stercoris, Collinsella tanakaei, Coprobacillus cateniformis, Coprobacter fastidiosus, Coprococcus catus, Coprococcus comes, Coprococcus eutactus, Corynebacterium ammoniagenes, Corynebacterium amycolatum amycolatum, Corynebacterium pseudodiphtheriticum, Cutibacterium acnesacnes, Dermabacter hominis, Desulfitobacterium hafniense, Desulfovibrio fairfieldensis, Desulfovibrio piger, Dialister succinatiphilus, Dielma fastidiosa, Dorea formicigenerans, Dorea longicatena, Dysgonomonas capnocytophagoides, Dysgonomonas gadei, Dysgonomonas mossii mossii, Edwardsiella tarda, Eggerthella lenta, Eisenbergiella tayi, Enorma massiliensis, Enterobacter aerogenes, Enterobacter asburiae, Enterobacter cancerogenus, Enterobacter cloacae, Enterobacter massiliensis, Enterococcus casseliflavus, Enterococcus durans, Enterococcus faecalis faecalis, Enterococcus faecium, Enterococcus flavescensflavescens, Enterococcus gallinarum, Enterococcus spp., Enterovibrio nigricans, Erysipelatoclostridium ramosum, Escherichia coli, Escherichia spp., Eubacterium biforme, Eubacterium dolichum, Eubacterium hallii, Eubacterium limosum, Eubacterium ramulus, Eubacterium rectale, Eubacterium siraeum), Eubacterium ventriosum (Euba Exiguobacterium ventriosum, Exiguobacterium marinum, Exiguobacterium undae, Faecalibacterium cf, Faecalibacterium prausnitzii, Faecalitalea cylindroides, Ferrimonas balearica, Finegoldia magna, Flavobacterium daejeonense, Flavonifractor plautii, Fusicatenibacter saccharivorans saccharivorans, Fusobacterium gonidiaformans, Fusobacterium mortiferum, Fusobacterium necrophorum, Fusobacterium nucleatum, Fusobacterium periodonticum, Fusobacterium spp., Fusobacterium ulcerans, Fusobacterium varium, Gallibacterium anatis, Gemmiger formicilis, Gordonibacter pamelaeae), Hafnia alvei, Helicobacter bilis, Helicobacter bilis, Helicobacter canadensis, Helicobacter caniscanis, Helicobacter cinaedi, Helicobacter macacae, Helicobacter pametensis, Helicobacter pullorum, Helicobacter pylori, Helicobacter rodentium, Helicobacter winghamensis, Herbaspirillum massiliense, Holdemanella biformis, Holdemania fdiformis, Holdemania filiformis, Holdemania masiliensis massiliensis, Holdemania filiformis, Hungatella hathewayi, Intestinibacter bartlettii, Intestinimonas butyriciproducens, Klebsiella oxytoca, Klebsiella pneumoniae, Kurthia massiliensis, Lachnospira pectinoschiza, Lactobacillus acidophilus, Lactobacillus amylolyticus, Lactobacillus animalis animalis), Lactobacillus antri, Lactobacillus brevisbrevis, Lactobacillus buchneri, Lactobacillus casei, Lactobacillus curvatus, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus hilgardii, Lactobacillus iners, Lactobacillus intestinalis, Lactobacillus johnsonii johnsonii, Lactobacillus murinus, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus ruminis, Lactobacillus sakei, Lactobacillus salivarius, Lactobacillus ultunensis, Lactobacillus vaginalis, Lactobacillus plantarum subsp. subsp.), Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Listeria greyigrayi, Listeria nocua, Mannheimia granulomatis, Marvinbryantia formatexigens, Megamonas funiformis, Megamonas hypermegale, Methanobrevibacter smithii, Methanobrevibacter smithiiFl, Micrococcus luteus, Microvirgula aerodenitrificans, Mitsuokella jalaludinii, Mitsuokella marutoashida multacida, Mollicutes bacterium, Murimonas intestini, Neisseria macacae, Nitriliruptor alkaliphilus, Oceanobacillus massiliensis, Odoribacter laneus, Odoribacter splanchnicus, Ornithobacterium rhinotracheale, Oxalobacter formigenes, Paenibacillus barengoldii barengoltzii, Paenibacillus chitinolyticus, Paenibacillus lautus, Paenibacillus motobuensismotobuensis, Paenibacillus senegalensis, Paenisporosarcina quisquiliarum, Parabacteroides distasonis, Parabacteroides goldsteinii, Parabacteroides gordonii, Parabacteroides johnsonii, Parabacteroides merdae, Paraprevotella xylaniphila, Parasutterella excrementihominis, Parvimonas micra micra, Pediococcus acidilactici, Peptoclostridium difficile, Peptoniphilus harei, Peptoniphilus obesi, Peptoniphilus senegalensis, Peptoniphilus timonensis, Phascolarctobacterium succinatutens, Porphyromonas asaccharolytica, Porphyromonas uenonis, Prevotella veroniae baroniae, Prevotella bivia, Prevotella copri, Prevotella dentalisPrevotella dentalis, Prevotella micans, Prevotella multisaccharivorax, Prevotella oralis, Prevotella salivae, Prevotella stercorea, Prevotella veroralis, Propionibacterium acnes, Propionibacterium avidum, Propionibacterium freudenreichii, Propionimicrobium lymphophilum, Proteus mirabilis mirabilis, Proteus penneri ATCC, Providencia alcalifaciens, Providencia rettgeri, Providencia rustigianii, Providencia stuartii, Pseudoflavonifractor capillosus, Pseudomonas aeruginosa, Pseudomonas luteola, Ralstonia pickettii, Rheinheimera perlucida, Rheinheimera texasensis, Riemerella columbina columbina), Romboutsia lituseburensis, Roseburia faecis, Roseburia intestinalisintestinalis, Roseburia inulinivorans, Ruminococcus bicirculans, Ruminococcus bromii, Ruminococcus callidus, Ruminococcus champanellensis, Ruminococcus faecis, Ruminococcus gnavus, Ruminococcus lactaris, Ruminococcus obeum, Ruminococcus spp., Ruminococcus turkis, Ruminococcus torques, Sarcina ventriculi, Sellimonas intestinalis, Senegalimassilia anaerobia, Shigella sonnei, Slackia piriformis, Staphylococcus epidermidis, Staphylococcus lentus, Staphylococcus nepalensis, Staphylococcus pseudintermedius, Staphylococcus xylosus, Stenotrophomonas maltophilia maltophilia, Streptococcus agalactiae, Streptococcus anginosus, Streptococcus australisaustralis, Streptococcus caballi, Streptococcus castoreus, Streptococcus didelphis, Streptococcus equinus, Streptococcus gordonii, Streptococcus henryi, Streptococcus hyovaginalis, Streptococcus infantarius, Streptococcus infantis, Streptococcus lutetiensis lutetiensis, Streptococcus merionis, Streptococcus Streptococcus mitis, Streptococcus mutans, Streptococcus oralis, Streptococcus ovis, Streptococcus parasanguinis, Streptococcus plurextorum, Streptococcus porci, Streptococcus pyogenes, Streptococcus salivarius, Streptococcus sobrinus, Streptococcus thermophilus thermophilus, Streptococcus thoraltensis, Streptomyces albus, Subdoligranulum variabile, Succinatimonas hippei, Sutterella parvirubra, Sutterella wadsworthensis, Terrisporobacter glycolicus, Terrisporobacter mayombei, Thalassobacillus devorans, Timonella senegalensis, Turicibacter sanguinis sanguinis, unknown genus, unknown genus, Varibaculum cambriense, Veillonella atypica, Veillonella dispardispar, Veillonella parvula, Vibrio cincinnatiensis, Virgibacillus salexigens, Weissella paramesenteroides, and Weissella paramesenteroides ATCC.

[0104] In other embodiments, the target bacterial cells are those commonly found in the skin microbiome and are preferably selected from the group consisting of Acetobacter farinalis, Acetobacter malorum, Acetobacter orleanensis, Acetobacter sicerae, Achromobacter anxifer, Achromobacter denitrificans, Achromobacter marplatensis, Achromobacter spanius, Achromobacter xylosoxidans subsp. xylosoxidans, Acidovorax conjaci, and the like. konjaci, Acidovorax radicis, Acinetobacter johnsonii, Actinomadura citrea, Actinomadura coerulea, Actinomadura fibrosa, Actinomadura fulvescens, Actinomadura jiaoheensis, Actinomadura luteofluorescens, Actinomadura mexicana, Actinomadura nitritigenes, Actinomadura verrucosospora, Actinomadura yumaensis, Actinomyces odontolyticus, Actinomycetospora atypicaatypica, Actinomycetospora corticicola, Actinomycetospora rhizophila, Actinomycetospora rishiriensis, Aeromonas australiensis, Aeromonas bestiarum, Aeromonas bivalvium, Aeromonas encheleia, Aeromonas eucrenophila, Aeromonas hydrophila subsp. hydrophila, Aeromonas piscicola piscicola, Aeromonas popoffii, Aeromonas rivuli, Aeromonas salmonicida subsp. pectinolytica, Aeromonas salmonicida subsp. smithia, Amaricoccus kaplicensis, Amaricoccus veronensis, Aminobacter aganoensis, Aminobacter ciceronei, Aminobacter lissarensis, Aminobacter niigataensis niigataensis, Ancylobacter polymorphus, Anoxybacillus flavithermus subsp.yunnanensis, Aquamicrobium aerolatum, Archangium gephyra, Archangium gephyra, Archangium minus, Archangium violaceum, Arthrobacter viscosus, Bacillus anthracis, Bacillus australimaris, Bacillus drentensis, Bacillus mycoides, Bacillus pseudomycoides, Bacillus pumilus, Bacillus safensis safensis, Bacillus vallismortis, Bosea thiooxidans, Bradyrhizobium huanghuaihaiense, Bradyrhizobium japonicum, Brevundimonas aurantiaca, Brevundimonas intermedia, Burkholderia aspalathi, Burkholderia choica, Burkholderia cordobensis, Burkholderia diffusa, Burkholderia insulsa, Burkholderia rhynchosiae, Burkholderia terrestris, Burkholderiaudeis, Buttiauxella gaviniae, Caenimonas terrestristerrae, Capnocytophaga gingivalis, Chitinophaga dinghuensis, Chryseobacterium gleum, Chryseobacterium greenlandense, Chryseobacterium jejuense, Chryseobacterium piscium, Chryseobacterium sediminis, Chryseobacterium tructae, Chryseobacterium ureilyticum, Chryseobacterium vietnamese vietnamense, Corynebacterium accolens, Corynebacterium afermentans subsp. lipophilum, Corynebacterium minutissimum, Corynebacterium sundsvallense, Cupriavidus metallidurans, Cupriavidus nantongensis, Cupriavidus necator, Cupriavidus pampae, Cupriavidus yeoncheonensis, Corynebacterium flaccumfaciens, Devosia epidermidihirudinis, Devosia riboflavina, Devosia riboflavina, Diaphorobacter oryzaeoryzae, Dietzia psychralcaliphila, Ensifer adhaerens, Ensifer americanus, Enterococcus malodoratus, Enterococcus pseudoavium, Enterococcus viikkiensis, Enterococcus xiangfangensis, Erwinia rhapontici, Falsirhodobacter halotolerans, Flavobacterium araucananum, Flavobacterium frigidimalis frigidimaris, Gluconobacter frateurii, Gluconobacter thailandicus, Gordonia alkanivorans, Halomonas aquamarina, Halomonas axialensis, Halomonas meridiana, Halomonas olivaria, Halomonas songnenensis, Halomonas variabilis, Herbaspirillum chlorophenolicum, Herbaspirillum frisingense, Herbaspirillum hiltneri hiltneri), Herbaspirillum huttiense subsp.putei, Herbaspirillum lusitanum, Herminiimonas fonticola, Hydrogenophaga intermedia, Hydrogenophaga pseudoflava, Klebsiella oxytoca, Kosakonia sacchari, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus modestisalitolerans, Lactobacillus plantarum subsp. argentoratensis, Lactobacillus xiangfangensis, Lechevalieria roselyniae, Lentzea albida, Lentzea californiensis, Leuconostoc carnosum, Leuconostoc citreum, Leuconostoc gelidum subsp. gasicomitatum, Leuconostoc mesenteroides subsp. suionicum, Luteimonas aestuarii, Lysobacter antibioticus, Lysobacter coeliac koreensis), Lysobacter oryzae (Lysobacter oryzae), Magnetospirillum moscoviense, Marinomonasalcarazii, Marinomonas primoryensis, Massilia aurea, Massilia jejuensis, Massilia kyonggiensis, Massilia timonae, Mesorhizobium acaciae, Mesorhizobium qingshengii, Mesorhizobium shonense, Methylobacterium haplocladii, Methylobacterium platani, Methylobacterium pseudosasicola, Methylobacterium zatmanii zatmanii, Microbacterium oxydans, Micromonospora chaiyaphumensis, Micromonospora chalcea, Micromonospora citrea, Micromonospora coxensis, Micromonospora echinofusca, Micromonospora halophytica, Micromonospora kangleipakensis, Micromonospora maritima, Micromonospora nigra, Micromonospora purpureochromogenes, Micromonospora rhizosphaerae rhizosphaerae, Micromonospora saelicesensis, Microvirga subterraneasubterranea, Microvirga zambiensis, Mycobacterium alvei, Mycobacterium avium subsp. silvaticum, Mycobacterium colombiense, Mycobacterium conceptionense, Mycobacterium conceptionense, Mycobacterium farcinogenes, Mycobacterium fortuitum subsp. fortuitum, Mycobacterium goodii, Mycobacterium insubricum insubricum, Mycobacterium llatzerense, Mycobacterium neoaurum, Mycobacterium neworleansense, Mycobacterium obuense, Mycobacterium peregrinum, Mycobacterium saopaulense, Mycobacterium Mycobacterium septicum, Mycobacterium setense, Mycobacterium smegmatis, Neisseria subflava, Nocardia lijiangensis, Nocardia thailandica, Novosphingobium barchaimii, Novosphingobium lindaniclasticum, Novosphingobium lindaniclasticum, Novosphingobium mathurense, Ochrobactrum pseudogrignonense, Oxalicibacterium solurbis, Paraburkholderia glaceri glathei, Paraburkholderia humi, Paraburkholderia phenazinium, Paraburkholderia phytofirmans, Paraburkholderia sordidicola, Paraburkholderia terricola, Paraburkholderia xenovorans, Paracoccus laeviglucosivorans, Patuli-libacter ginsengiterrae, Polymorphospora rubra, Porphyrobacter colymbi, Prevotella jejuni jejuni), Prevotella melaninogenica (Prevotellamelaninogenica, Propionibacterium acnes subsp. elongatum, Proteus vulgaris, Providencia rustigianii, Pseudoalteromonas agarivorans, Pseudoalteromonas atlantica, Pseudoalteromonas paragorgicola, Pseudomonas asplenii, Pseudomonas asuensis, Pseudomonas benzenivorans, Pseudomonas cannabina cannabina, Pseudomonas cissicola, Pseudomonas congelans, Pseudomonas costantinii, Pseudomonas ficuserectae, Pseudomonas frederiksbergensis, Pseudomonas graminis, Pseudomonas jessenii, Pseudomonas koreensis, Pseudomonas koreensis, Pseudomonas kunmingensis, Pseudomonas marginalis marginalis, Pseudomonas mucidolens, Pseudomonas panacis, Pseudomonas plecoglossidaplecoglossicida, Pseudomonas poae, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas reinekei, Pseudomonas rhizosphaerae, Pseudomonas seleniipraecipitans, Pseudomonas umsongensis, Pseudomonas zhaodongensis, Pseudonocardia alaniniphila, Pseudonocardia ammonioxydans ammonioxydans, Pseudonocardia autotrophica, Pseudonocardia kongjuensis, Pseudonocardia yunnanensis, Pseudorhodoferax soli, Pseudoxanthomonas daejeonensis, Pseudoxanthomonas indica, Pseudoxanthomonas kaohsiungensis, Psychrobacter aquaticus, Psychrobacter arcticus, Psychrobacter sere celer), Psychrobacter marincola, Psychrobacter nivimaris, Psychrobacter okhotsukensisokhotskensis), Psychrobacter okhotskensis, Psychrobacter piscatorii, Psychrobacter pulmonis, Ramlibacter ginsenosidimutans, Rheinheimera japonica, Rheinheimera muenzenbergensis, Rheinheimera soli, Rheinheimera tangshanensis, Rheinheimera texasensis, Rheinheimera tilapiae, Rhizobium alamii, Rhizobium azibense, Rhizobium vinae binae, Rhizobium daejeonense, Rhizobium endophyticum, Rhizobium etli, Rhizobium fabae, Rhizobium freirei, Rhizobium gallicum, Rhizobium loessense, Rhizobium sophoriradicis, Rhizobium taibaishanense, Rhizobium vallis, Rhizobium vignae, Rhizobium yanglingense, Rhodococcus baiconulensis baikonurensis), Rhodococcus enclensis, Rhodoferax saidenbachensis, Rickettsia canadensis, Rickettsia heilongjiangensis, Rickettsia honeihonei, Rickettsia raoultii, Roseateles aquatilis, Roseateles aquatilis, Salmonella enterica subsp. salamae, Serratia ficaria, Serratia myotis, Serratia vespertilionis, Shewanella aestuarii, Shewanella decolorationis, Sphingobium amiense, Sphingobium baderi, Sphingobium barthaii, Sphingobium chlorophenolicum, Sphingobium cupriresistens, Sphingobium czechense, Sphingobium fuliginis, Sphingobium indicum, Sphingobium indicum, Sphingobium japonicum, Sphingobium lactosutens, Sphingomonas dokdonensis, Sphingomonas pseudosanguinis, Sphingopyxis chilensis, Sphingopyxis fribergensis, Sphingopyxis granuli, Sphingopyxis indica indica), Sphingopyxis witflariensis, Staphylococcus agnetis, Staphylococcus aureus subsp. aureusaureus subsp. aureus, Staphylococcus epidermidis, Staphylococcus hominis subsp. novobiosepticus, Staphylococcus nepalensis, Staphylococcus saprophyticus subsp. bovis, Staphylococcus sciuri subsp. carnaticus, Streptomyces caeruleus, Streptomyces canarius, Streptomyces capoamus capoamus, Streptomyces ciscaucasicus, Streptomyces griseorubiginosus, Streptomyces olivaceoviridis, Streptomyces panaciradicis, Streptomyces phaeopurpureus, Streptomyces pseudovenezuelae, Streptomyces resistomycificus, Tianweitania sediminis, Tsukamurella paurometabola, Variovorax guangxiensis, Vogesella alkaliphila), Xanthomonas arboricola, Xanthomonas axonopodisaxonopodis, Xanthomonas cassavae, Xanthomonas cucurbitae, Xanthomonas cynarae, Xanthomonas euvesicatoria, Xanthomonas fragariae, Xanthomonas gardneri, Xanthomonas perforans, Xanthomonas pisi, Xanthomonas populi, Xanthomonas vasicola, Xenophilus aerolactosus aerolatus, Yersinia nurmii, Abiotrophia defectiva, Acidocella aminolytica, Acinetobacter guangdongensis, Acinetobacter parvus, Acinetobacter radioresistens, Acinetobacter soli, Acinetobacter variabilis, Actinomyces cardiffensis, Actinomyces dentalis, Actinomyces europaeus, Actinomyces gerenseriae gerencseriae, Actinomyces graevenitzii, Actinomyces haliotis, Actinomyces johnsoniiActinomyces johnsonii, Actinomyces massiliensis, Actinomyces meyeri, Actinomyces mayeri, Actinomyces naeslundii, Actinomyces neuii subsp. anitratus, Actinomyces odontolyticus, Actinomyces oris, Actinomyces turicensis, Actinomycetospora corticicola, Actinotignum schaalii, Aerococcus christensenii christensenii, Aerococcus urinae, Aeromicrobium flavum, Aeromicrobium massiliense, Aeromicrobium tamlense, Aeromonas sharmana, Aggregatibacter aphrophilus s), Aggregatibacter segnis, Agrococcus baldri, Albibacter methylovorans, Alcaligenes faecalis subsp. faecalis, Algoriphagus ratkowskyi, Alkalibacterium olivapovliticus, Alkalibacterium pelagium, Alloprevotella rava, Alsobacter metallidurans, Amaryllidaceus capricensis kaplicensis, Amaricoccus veronensis, Anaerococcus hydrogenalis, Anaerococcus lactolyticus, Anaerococcus murdochii, Anaerococcus octavius, Anaerococcus prevotii, Anaerococcus vaginalis, Aquabacterium citratiphilum, Aquabacterium olei, Aquabacterium olei, Aquabacterium parvum parvum, Aquincola tertiaricarbonis, Arcobacter venerupis, Arsenicicoccus bolidensis, Arthrobacter lusicusrussicus, Asticcacaulis excentricus, Atopobium deltae, Atopobium parvulum, Atopobium rimae, Atopobium vaginae, Aureimonas altamirensis, Aureimonas rubiginis, Azospira oryzae, Azospirillum oryzae, Bacillus circulans, Bacillus drentensis, Bacillus fastidiosus, Bacillus lehensis, Bacillus oceanisediminis, Bacillus rhizosphaerae, Bacteriovorax stolpii, Bacteroides coagulan, Bacteroides dorei, Bacteroides fragilis, Bacteroides ovatus, Bacteroides stercoris, Bacteroides uniformis, Bacteroides vulgatus, Bdellovibrio bacteriovorus, Bdellovibrio exovorus, Belnapia moabensis, Belnapia soli, Blautia hansenii, Blautia obeum, Blautia wechslerwexlerae, Bosea lathyri, Brachybacterium fresconis, Brachybacterium muris, Brevibacterium ammoniilyticum, Brevibacterium casei, Brevibacterium epidermidis, Brevibacterium iodinum, Brevibacterium luteolum, Brevibacterium paucivorans, Brevibacterium pityocampae, Brevibacterium sanguinis sanguinis, Brevundimonas albigilva, Brevundimonas diminuta, Brevundimonas vancanneytii, Caenimonas terrae, Calidifontibacter indicus, Campylobacter concisus, Campylobacter gracilis, Campylobacter hominis, Campylobacter rectus, Campylobacter showae, Campylobacter ureolyticus, Capnocytophaga gingivalis gingivalis), Capnocytophaga leadbetteri, Capnocytophaga ochracea, Capnocytophaga sp.sputigena, Cardiobacterium hominis, Cardiobacterium valvarum, Carnobacterium divergens, Catonella morbi, Caulobacter henricii, Cavicella subterranea, Cellulomonas xylanilytica, Cellvibrio vulgaris, Chitinimonas taiwanensis, Chryseobacterium arachidis, Chryseobacterium deceongens daecheongense, Chryseobacterium formosense, Chryseobacterium formosense, Chryseobacterium greenlandense, Chryseobacterium indologenes, Chryseobacterium piscium, Chryseobacterium rigui, Chryseobacterium solani, Chryseobacterium taklimakanense, Chryseobacterium ureilyticum, Chryseobacterium ureilyticum, Chryseobacterium jiae zeae), Chryseomicrobium aureum, Cloacibacterium haliotis, Cloacibacterium normensisnormanense, Cloacibacterium normanens, Collinsella aerofaciens, Comamonas denitrificans, Comamonas terrigena, Corynebacterium accolens, Corynebacterium afermentans subsp. lipophilum, Corynebacterium ammoniagenes, Corynebacterium amycolatum, Corynebacterium auricosum aurimucosum, Corynebacterium aurimucosum, Corynebacterium coyleae, Corynebacterium durum, Corynebacterium freiburgense, Corynebacterium glaucum, Corynebacterium glyciniphilum, Corynebacterium imitans, Corynebacterium jeikeium, Corynebacterium kroppenstedtii, Corynebacterium lipophylloflavum lipophiloflavum, Corynebacterium massiliense, Corynebacterium mastitidis, Corynebacterium mattorcotiimatruchotii, Corynebacterium minutissimum, Corynebacterium mucifaciens, Corynebacterium mustelae, Corynebacterium mycetoides, Corynebacterium pyruviciproducens, Corynebacterium simulans, Corynebacterium singulare, Corynebacterium sputi, Corynebacterium suicordis suicordis, Corynebacterium tuberculostearicum, Corynebacterium tuberculostearicum, Corynebacterium ureicelerivorans, Corynebacterium variabile, Couchioplanes caeruleus subsp. caeruleus, Cupriavidus metallidurans, Curtobacterium herbarum, Dechloromonas agitata, Deinococcus actinosclerus actinosclerus, Deinococcus antarcticus, Deinococcus caeni, Deinococcus ficusficus, Deinococcus geothermalis, Deinococcus radiodurans, Deinococcus wulumuqiensis, Deinococcus xinjiangensis, Dermabacter hominis, Dermabacter vaginalis, Dermacoccus nishinomiyaensis, Desemzia incerta, Desertibacter roseus, Dialister invisus, Dialister micraerophilus, Dialister propionicifaciens propionicifaciens, Dietzia aurantiaca, Dietzia cercidiphylli, Dietzia timorensis, Dietzia timorensis, Dolosigranulum pigrum, Eikenella corrodens, Elizabethkingia miricola, Elstera litoralis, Empedobacter brevis, Enhydrobacter aerosaccus, Enterobacter shanfangensis xiangfangensis), Enterococcus aquimarinus, Enterococcus faecalisfaecalis, Enterococcus olivae, Erwinia rhapontici, Eubacterium eligens, Eubacterium infirmum, Eubacterium rectale, Eubacterium saphenum, Eubacterium sulci, Exiguobacterium mexicanum, Facklamia tabacinasalis, Falsirhodobacter halotolerans halotolerans, Finegoldia magna, Flavobacterium cutihirudinis, Flavobacterium lindanitolerans, Flavobacterium resistens, Friedmanniella capsulata ulata, Fusobacterium nucleatum subsp. polymorphum, Gemella haemolysans, Gemella morbillorum, Gemella palaticanis, Gemella sanguinis, Gemmobacter aquaticus, Gemmobacter caeni, Gordonia jinhuaensis, Gordonia kroppenstedtii, Gordonia polyisoprenivorans, Gordonia polyisoprenivorans, Granulicatella adiasens adiacens, Granulicatella elegans, Haemophilus parainfluenzae, Haemophilus sputorum, Halomonas sulfidaeris, Herpetosiphon aurantiacus, Hydrocarboniphaga effusa, Idiomarina maris, Janibacter anophelis, Janibacter hoylei, Janibacter indicus, Janibacter rimosus limosus, Janibacter melonis, Jeotgalicoccus halophilus, Jonquetella anthropi, Kaistiageumhonensis, Kingella denitrificans, Kingella oralis, Klebsiella oxytoca, Knoellia aerolata, Knoellia locipacati, Kocuria atrinae, Kocuria carniphila, Kocuria kristinae, Kocuria palustris, Kocuria turfanensis, Lachnoanaerobaculum saburreum, Lachnoanaerobaculum sabreum, Lactobacillus crispatus Lactococcus crispatus, Lactobacillus iners, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis, Lactococcus piscium, Lapillicoccus jejuensis, Lautropia mirabilis, Legionella beliardensis, Leptotrichia buccalis, Leptotrichia goodfellowii, Leptotrichia hofstadii, Leptotrichia hongkongensis, Leptotrichia sha'ii shahii), Leptotrichia trevisanii, Leptotrichia wadei, Luteimonasterricola, Lysinibacillus fusiformis, Lysobacter spongiicola, Lysobacter xinjiangensis, Macrococcus caseolyticus, Marmoricola pocheonensis, Marmoricola scoriae, Massilia alkalitolerans, Massilia alkalitolerans, Massilia aurea, Massilia plicata, Massilia timonae, Megamonas rupellensis, Meiothermus silvanus, Methylobacterium tangquense dankookense, Methylobacterium goesingense, Methylobacterium gesingens, Methylobacterium isbiliense, Methylobacterium jeotgali, Methylobacterium oxalidis, Methylobacterium platani, Methylobacterium pseudosasicola, Methyloversatilis universalis, Microbacterium foliorum, Microbacterium hydrosarmales hydrothermale), Microbacterium hydrothermale, Microbacterium lacticum, Microbacterium lacticum, Microbacterium lavenniformanslaevaniformans, Microbacterium paludicola, Microbacterium petrolearium, Microbacterium phyllosphaerae, Microbacterium resistens, Micrococcus antarcticus, Micrococcus cohnii, Micrococcus luteus, Micrococcus lylae, Micrococcus terreus, Microlunatus aurantiacus, Micropruina glycogenica, Microvirga erilata aerilata, Microvirga erilata, Microvirga subterranea, Microvirga vignae, Microvirga zambiensis, Microvirgula aerodenitrificans, Mogibacterium timidum, Moraxella atlantae, Moraxella catarrhalis, Morganella morganii subsp. morganii, Morganella psychrotolerans, Murdochiella asaccharolytica, Mycobacterium asiaticum asiaticum), Mycobacterium chubuense, Mycobacteriumcrocinum, Mycobacterium gadium, Mycobacterium holsaticum, Mycobacterium iranicum, Mycobacterium longobardum, Mycobacterium neoaurum, Mycobacterium neoaurum, Mycobacterium obuense, Negativicoccus succinicivorans, Neisseria bacilliformis, Neisseria oralis, Neisseria sicca, Neisseria subflava subflava), Nesterenkonia lacusekhoensis, Nesterenkonia rhizosphaerae, Nevskia persephonica, Nevskia ramosa, Niabella yanshanensis, Niveibacterium umoris, Nocardia niwae, Nocardia thailandica, Nocardioides agariphilus, Ocardioides dilutus, Nocardioides ganghwensis, Nocardioides hwasunensis, Nocardioides nanhaiensis, Nocardioides sediminis, Nosocomiicoccus ampullae, Noviherbaspirillum malthae, Novosphingobium lindaniclasticum (Novosphingobiumlindaniclasticum, Novosphingobium rosa, Ochrobactrum rhizosphaerae, Olsenella uli, Ornithinimicrobium murale, Ornithinimicrobium tianjinense, Oryzobacter terrae, Ottowia beijingensis, Paenalcaligenes suwonensis, Paenibacillus agaridevorans, Paenibacillus phoenicis, Paenibacillus xylanexedens, Paludibacterium yongneupense, Pantoea cypripedii, Parabacteroides distasonis, Paraburkholderia andropogonis, Paracoccus alcaliphilus, Paracoccus angustae, Paracoccus kocurii, Paracoccus laeviglucosivorans, Paracoccus sediminis diminis, Paracoccus sphaerophysae, Paracoccus yeei, Parvimonas micra, Parviterribacter multiflagellatus, Patulibacter ginsengiterrae, Pedobacter aquatilis, Pedobacter ginsengisoli, Pedobacter xixiisoli, Peptococcus niger, Peptoniphilus coxii, Peptoniphilus gorbachii, Peptoniphilus harei, Peptoniphilus coenoenenniae koenoeneniae, Peptoniphilus lacrimalis, Peptostreptococcus anaerobius, Peptostreptococcus stomatis, Phascolarctobacterium faecium, Phenylobacterium haematophilum, Phenylobacterium kunshanense, Pluralibacter gergoviae, Polymorphobacter multimanifer, Porphyromonas bennonis, Porphyromonas endodontalis endodontalis, Porphyromonas gingivalis, Porphyromonas gingivicanis, Porphyromonaspasteri, Porphyromonas pogonae, Porphyromonas somerae, Povalibacter uvarum, Prevotella aurantiaca, Prevotella baroniae, Prevotella bivia, Prevotella buccae, Prevotella buccalis, Prevotella copri, Prevotella corporis, Prevotella denticola, Prevotella enoeca, Prevotella histicola histicola, Prevotella intermedia, Prevotella jejuni, Prevotella jejuni, Prevotella maculosa, Prevotella melaninogenica, Prevotella melaninogenica, Prevotella micans, Prevotella multiformis, Prevotella nanceiensis, Prevotella nigrescens, Prevotella oris, Prevotella oulorum, Prevotella parens pallens), Prevotella pleuritidis, Prevotella saccharolytica, Prevotella salivariussalivae, Prevotella shahii, Prevotella timonensis, Prevotella veroralis, Propionibacterium acidifaciens, Propionibacterium acnes subsp. acnes, Propionibacterium acnes subsp. acnes, Propionibacterium acnes subsp. elongatum, Propionibacterium granulosum, Propionimicrobium lymphophilum lymphophilum, Propionispira arcuata, Pseudokineococcus lusitanus, Pseudomonas aeruginosa, Pseudomonas chengduensis, Pseudonocardia benzenivorans, Pseudorhodoplanes sinuspersici, Psychrobacter sanguinis, Ramlibacter ginsenosidimutans, Rheinheimera aquimaris, Rhizobium alvei, Rhizobium daejonens daejeonense, Rhizobium larrymoorei, Rhizobium rhizoryzae, Rhizobium soli, Rhizobium taibaishanense, Rhizobium vignae, Rhodanobacterglycinis, Rhodobacter veldkampii, Rhodococcus enclensis, Rhodococcus fascians, Rhodococcus fascians, Rhodovarius lipocyclicus, Rivicola pingtungensis, Roseburia inulinivorans, Rosenbergiella nectarea, Roseomonas aerilata, Roseomonas aquatica, Roseomonas mucosa, Roseomonas rosea rosea, Roseomonas vinacea, Rothia aeria, Rothia amarae, Rothia dentocariosa, Rothia endophytica, Rothia mucilaginosa, Rothia nasimurium, Rubellimicrobium mesophilum, Rubellimicrobium roseum, Rubrobacter bracarensis, Rudaea cellulosilytica, Ruminococcus gnavus, Runella zeae, Saccharopolyspora rectivirgra rectivirgula, Salinicoccus qingdaonensis, Scardovia wiggsiae, Sediminibacterium ginsengisoli, Selenomonas artemidisartemidis, Selenomonas infelix, Selenomonas noxia, Selenomonas sputigena, Shewanella aestuarii, Shuttleworthia satelles, Simonsiella muelleri, Skermanella aerolata, Skermanella stibiiresistens, Slackia exigua, Smaragdicoccus niigatensis, Sneathia sanguinegens, Solirubrobacter soli soli, Sphingobacterium caeni, Sphingobacterium daejeonense, Sphingobacterium hotanense, Sphingobacterium kyonggiense, Sphingobacterium multivorum, Sphingobacterium nematocida, Sphingobacterium spiritivorum, Sphingobium amiense, Sphingobium indicum, Sphingobium lactosutens, Sphingobium subterranean subterraneum, Sphingomonas abaci, Sphingomonas aestuarii, Sphingomonas canadensiscanadensis, Sphingomonas daechungensis, Sphingomonas dokdonensis, Sphingomonas echinoides, Sphingomonas fonticola, Sphingomonas fonticola, Sphingomonas formosensis, Sphingomonas gei, Sphingomonas hankookensis, Sphingomonas hankookensis, Sphingomonas koreensis, Sphingomonas kyeonggiensis, Sphingomonas laterariae laterariae, Sphingomonas mucosissima, Sphingomonas oligophenolica, Sphingomonas pseudosanguinis, Sphingomonas sediminicola, Sphingomonas yantingensis, Sphingomonas yunnanensis, Sphingopyxis indica, Spirosoma rigui, Sporacetigenium mesophilum, Sporocytophaga myxococcoides, Staphylococcus auricularis auricularis, Staphylococcus epidermidis, Staphylococcus epidermidisepidermidis, Staphylococcus hominis subsp. novobiosepticus, Staphylococcus lugdunensis, Staphylococcus pettenkoferi, Stenotrophomonas koreensis, Stenotrophomonas rhizophila, Stenotrophomonas rhizophila, Streptococcus agalactiae, Streptococcus canis, Streptococcus cristatus, Streptococcus gordonii gordonii, Streptococcus infantis, Streptococcus intermedius, Streptococcus mutans, Streptococcus oligofermentans, Streptococcus oralis, Streptococcus sanguinis, Streptomyces iconiensis, Streptomyces yanglinensis, Tabrizicola aquatica, Tahibacter caeni, Tannerella forsythia, Tepidicella xavieri, Tepidimonas fonticaldi, Terracoccus luteus, Tessaracoccus flavescens, Thermus thermophilusthermophilus, Tianweitania sediminis, Tianweitania sediminis, Treponema amylovorum, Treponema denticola, Treponema lecithinolyticum, Treponema medium, Turicella otitidis, Turicibacter sanguinis, Undibacterium oligocarboniphilum, Undibacterium squillarum, Vagococcus salmoninarum, Varibaculum cambriense, Vibrio metschnikoffii metschnikovii, Xanthobacter tagetidis, Xenophilus aerolatus, Xenophilus arseniciresistens, Yimella lutea, Zimmermannella alba, Zimmermannella bifida da), and Zoogloea caeni.

[0105] In other embodiments, the target bacterial cells are those commonly found in the vaginal microflora, preferably Acinetobacter antiviralis, Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter johnsonii, Actinobaculum massiliense, Actinobaculum schaalii, Actinomyces europaeus, Actinomyces graevenitzii, Actinomyces israelii, Actinomyces mayeri, Actinomyces naeslundii, Actinomyces Actinomyces naeslundii, Actinomyces neuii, Actinomyces odontolyticus, Actinomyces turicensis, Actinomyces urogenitalis, Actinomyces viscosus, Aerococcus christensenii, Aerococcus urinae, Aerococcus viridans, Aeromonas encheleia, Aeromonas salmonicida, Afipia masciliensis massiliensis, Agrobacterium tumefaciens, Algoriphagus aquatilis, Aliivibrio wodaniswodanis, Alistipes finegoldii, Alloiococcus otitis, Alloprevotella tannerae, Alloscardovia omnicolens, Altererythrobacter epoxidivorans, Ammoniphilus oxalaticus, Amnibacterium kyonggiense, Anaerococcus hydrogenalis, Anaerococcus lactolyticus, Anaerococcus murdochii, Anaerococcus obesiensis obesiensis, Anaerococcus prevotii, Anaerococcus tetradius, Anaerococcus vaginalis, Anaeroglobus geminatus, Anoxybacillus pushchinoensis, Aquabacterium parvum, Arcanobacterium phocae, Arthrobacter aurescens, Asticcacaulis excentricus, Atopobium minutum, Atopobium parvum parvulum, Atopobium rimae, Atopobium vaginae, Avibacterium gallinarum, Bacillus acidicolaacidicola, Bacillus atrophaeus, Bacillus cereus, Bacillus cibi, Bacillus coahuilensis, Bacillus gaemokensis, Bacillus methanolicus, Bacillus oleronius, Bacillus pumilus, Bacillus shackletonii, Bacillus sporothermodurans, Bacillus subtilis, Bacillus wakoensis wakoensis, Bacillus weihenstephanensis, Bacteroides barnesiae, Bacteroides coagulan, Bacteroides dorei, Bacteroides faecis, Bacteroides forsythus, Bacteroides fragilis, Bacteroides nordii, Bacteroides ovatus, Bacteroides salyersiae, Bacteroides stercoris, Bacteroides uniformis uniformis, Bacteroides vulgatus, Bacteroides xylanisolvens, Bacteroides zoogleoformanszoogleoformans, Barnesiella viscericola, Bhargavaea cecembens, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium dentium, Bifidobacterium logum subsp. infantis, Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Bifidobacterium skaldovii scardovii, Bilophila wadsworthia, Blautia hydrogenotrophica, Blautia obeum, Blautia producta, Brachybacterium faecium, Bradyrhizobium japonicum, Brevibacterium mcbrellneri, Brevibacterium otitidis, Brevibacterium paucivorans, Bulleidia extructa, Burkholderia fungorum fungorum), Burkholderia phenoliruptix, Caldicellulosiruptor saccharolyticus (Caldicellulosiruptorsaccharolyticus, Caldimonas taiwanensis, Campylobacter gracilis, Campylobacter hominis, Campylobacter sputorum, Campylobacter ureolyticus, Capnocytophaga ochracea, Cardiobacterium hominis, Catonella morbi, Chlamydia trachomatis, Chlamydophila abortus, Chondromyces robustus robustus, Chryseobacterium aquaticum, Citrobacter youngae, Cloacibacterium normensis, Clostridium cavendishii, Clostridium colicanis, Clostridium jejuense, Clostridium perfringens, Clostridium ramosum, Clostridium sordellii, Clostridium viride, Comamonas terrigena, Corynebacterium aquaticum accolens, Corynebacterium appendicis, Corynebacterium koileaecoyleae, Corynebacterium glucuronolyticum, Corynebacterium glutamicum, Corynebacterium jeikeium, Corynebacterium kroppenstedtii, Corynebacterium lipophiloflavum, Corynebacterium minutissimum, Corynebacterium mucifaciens, Corynebacterium nuruki, Corynebacterium pseudogenitalium Corynebacterium pseudogenitalium, Corynebacterium pyruviciproducens, Corynebacterium singulare, Corynebacterium striatum, Corynebacterium tuberculostearicum, Corynebacterium xerosis, Cryobacterium psychrophilum, Corynebacterium flaccumfaciens, Cutibacterium acnes, Cutibacterium avidum avidum, Cytophaga xylanolytica, Deinococcus radiophilus, Delftiatsuruhatensis, Desulfovibrio desulfuricans, Dialister invisus, Dialister micraerophilus, Dialister pneumosintes, Dialister propionicifaciens, Dickeya chrysanthemi, Dorea longicatena, Eggerthella lenta, Eggerthia catenaformis, Eikenella corrodens, Enhydrobacter aerosax aerosaccus, Enterobacter asburiae, Enterobacter cloacae, Enterococcus avium, Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Enterococcus hirae, Erwinia persicina, Erwinia rhapontici, Erwinia toletana, Escherichia coli, Escherichia fergusonii, Eubacterium brachyura brachy, Eubacterium eligens, Eubacterium nodatum, Eubacterium rectale, Eubacterium safenumsaphenum, Eubacterium siraeum, Eubacterium sulci, Eubacterium yurii, Exiguobacterium acetylicum, Facklamia ignava, Faecalibacterium prausnitzii, Filifactor alocis, Finegoldia magna, Fusobacterium gonidiaformans, Fusobacterium nucleatum nucleatum, Fusobacterium periodonticum, Gardnerella vaginalis, Gemella asaccharolytica , Gemella bergeri, Gemella haemolysans, Gemella sanguinis, Geobacillus stearothermophilus, Geobacillus thermocatenulatus, Geobacillus thermoglucosidasius, Geobacter grbiciae, Granulicatella elegans, Haemophilus ducreyi, Haemophilus haemolyticus, Haemophilus parahaemolyticus parahaemolyticus, Haemophilus parainfluenzae, Hafnia alvei, Halomonas meridiana, Halomonas phoceae, Halomonas venusta, Herbaspirillum seropedicae, Janthinobacterium lividum, Jonquetella anthropi, Klebsiella granulomatis, Klebsiella oxytoca, Klebsiella pneumoniae, Lactobacillus acidophilus acidophilus, Lactobacillus amylovorus, Lactobacillus brevis, Lactobacillus koreohominiscoleohominis, Lactobacillus crispatus, Lactobacillus curvatus, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus kalixensis, Lactobacillus kefiranofaciens Lactobacillus kefiranofaciens, Lactobacillus kimchicus, Lactobacillus kitasatonis, Lactobacillus mucosae, Lactobacillus panis, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus pontis, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus altunensis ultunensis, Lactobacillus vaginalis, Lactococcus lactis, Leptotrichiabuccalis, Leuconostoc carnosum, Leuconostoc citreum, Leuconostoc garlicum, Leuconostoc lactis, Leuconostoc mesenteroides, Lysinimonas kribbensis, Mageeibacillus indolicus, Maribacter orientalis, Marinomonas protea, Marinospirillum insulare, Massilia timonae, Megasphaera elsdenii, Megasphaera micronuciformis micronuciformis, Mesorhizobium amorphae, Methylobacterium radiotolerans, Methylotenera versatilis, Microbacterium halophilum, Micrococcus luteus, Microterricola viridarii, Mobiluncus curtisii, Mobiluncus mulieris, Mogibacterium timidum, Moorella glycerini, Moraxella osloensis, Morganella morganii, Moriella indrigenes indoligenes, Murdochiella asaccharolytica, Mycoplasma albialvi, Mycoplasma genitalium, Mycoplasma hominis, Mycoplasma muris, Mycoplasma salivarium, Negativicoccus succinicivorans, Neisseria flava, Neisseria gonorrhoeae, Neisseria mucosa, Neisseria subflava, Nevskia ramosa, Nevskia soli, Nitriliruptor alkaliphilus, Odoribacter plankunnicus splanchnicus, Oligella urethralis, Olsenella uli, Paenibacillus amylolyticus, Paenibacillus humicus, Paenibacillus pabuli, Paenibacillus pasadenensis, Paenibacillus pini, Paenibacillus validus, Pantoea agglomerans, Parabacteroides merdae, Paraburkholderia caryophyllii caryophylli, Paracoccus yeei, Parastreptomyces abscessus, Parvimonas micra, Pectobacterium betabusculorumbetavasculorum, Pectobacterium carotovorum, Pediococcus acidilactici, Pediococcus ethanolidurans, Pedobacter alluvionis, Pedobacter wanjuense, Pelomonas aquatica, Peptococcus niger, Peptoniphilus asaccharolyticus, Peptoniphilus gorbachii, Peptoniphilus harei, Peptoniphilus indricus indolicus, Peptoniphilus lacrimalis, Peptoniphilus massiliensis, Peptostreptococcus anaerobius, Peptostreptococcus massiliae, Peptostreptococcus stomatis, Photobacterium angustum, Photobacterium frigidiphilum, Photobacterium phosphoreum, Porphyromonas asaccharolytica, Porphyromonas venonis bennonis, Porphyromonas catoniae, Porphyromonas endodontalisendodontalis, Porphyromonas gingivalis, Porphyromonas somerae, Porphyromonas uenonis, Prevotella amnii, Prevotella baroniae, Prevotella bergensis, Prevotella bivia, Prevotella buccae, Prevotella buccalis, Prevotella colorans, Prevotella copri, Prevotella corporis, Prevotella dentalis dentalis, Prevotella denticola, Prevotella disiens, Prevotella intermedia, Prevotella loescheii, Prevotella marshii, Prevotella melaninogenica, Prevotella micans, Prevotella nigrescens, Prevotella oris, Prevotella pleuritidis, Prevotella ruminicola, Prevotella shahii, Prevotella starcorea stercorea), Prevotella timonensis, Prevotella veroralisveroralis, Propionimicrobium lymphophilum, Proteus mirabilis, Pseudomonas abietaniphila, Pseudomonas aeruginosa, Pseudomonas amygdali, Pseudomonas azotoformans, Pseudomonas chlororaphis, Pseudomonas cuatrocienegasensis, Pseudomonas fluorescens, Pseudomonas fulva, Pseudomonas lutea lutea, Pseudomonas mucidolens, Pseudomonas oleovorans, Pseudomonas orientalis, Pseudomonas pseudoalcaligenes, Pseudomonas psychrophila, Pseudomonas putida, Pseudomonas synxantha, Pseudomonas syringae, Pseudomonas tolaasii, Pseudopropionibacterium propionicum, Rahnella aquatilis aquatilis, Ralstonia pickettii, Ralstonia solanacearum, Raoultella planticolaplanticola, Rhizobacter dauci, Rhizobium etli, Rhodococcus fascians, Rhodopseudomonas palustris, Roseburia intestinalis, Roseburia inulinivorans, Rothia mucilaginosa, Ruminococcus bromii, Ruminococcus gnavus, Ruminococcus torques, Sanguibacter keddieii, Sediminibacterium salmoneum salmoneum, Selenomonas bovis, Serratia fonticola, Serratia liquefaciens, Serratia marcescens, Shewanella algae, Shewanella amazonensis, Shigella boydii, Shigella sonnei, Slackia exigua, Sneathia amnii, Sneathia sanguinegens, Solobacterium moorei, Sorangium cellulosum, Sphingobium amiense, Sphingobium japonicum, Sphingobium yanoikuyae, Sphingomonas wittichii wittichii, Sporosarcina aquimarina, Staphylococcus aureus, Staphylococcus auricularis, Staphylococcus capitis, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus lugdunensis, Staphylococcus saprophyticus, Staphylococcus schleiferi Staphylococcus schleiferi, Staphylococcus simiae, Staphylococcus simulans, Staphylococcus warneri, Stenotrophomonas maltophiliamaltophilia, Stenoxybacter acetivorans, Streptococcus agalactiae, Streptococcus anginosus, Streptococcus australis, Streptococcus equinus, Streptococcus gallolyticus, Streptococcus infantis, Streptococcus intermedius, Streptococcus lutetiensis, Streptococcus marimammalium, Streptococcus mitis mitis, Streptococcus mutans, Streptococcus oralis, Streptococcus parasanguinis, Streptococcus phocae, Streptococcus pseudopneumoniae, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus thermophilus, Sutterella wadsworthensis, Tannerella forsythia, Terrahaemophilus aromaticivorans, Treponema denticola, Treponema maltophyllummaltophilum, Treponema parvum, Treponema vincentii, Trueperella bernardiae, Turicella otitidis, Ureaplasma parvum, Ureaplasma urealyticum, Varibaculum cambriense, Variovorax paradoxus, Veillonella atypica, Veillonella dispar, Veillonella montpellierensis, Veillonella parvula parvula, Virgibacillus proomii, Viridibacillus arenosi, Viridibacillus arvi, Weissella cibaria, Weissella soli, Xanthomonas campestris, Xanthomonas vesicatoria, Zobellia laminariae, and Zoogloea ramigera.

[0106] In one embodiment, the target bacterium is Escherichia coli.

[0107] In one embodiment, the target bacterium is Cutibacterium acnes, more particularly acne-associated Cutibacterium acnes from phylogroup IA1, or RT4, RT5, RT8, RT9, RT10, or clonal complexes (CC) CC1, CC3, and CC4, more particularly ST1, ST3, and ST4.

[0108] Thus, bacteriophages used to prepare bacterial viral particles, as well as bacterial viral particles, may target (e.g., specifically target) bacterial cells from any one or more of the aforementioned genera and / or species of bacteria to specifically deliver a payload.

[0109] In one embodiment, the target bacteria is a pathogenic bacterium. The targeted bacteria can be a virulent bacterium.

[0110] The target bacterium may preferably be an antibacterial-resistant bacterium selected from the group consisting of extended-spectrum beta-lactamase-producing (ESBL) Escherichia coli, ESBL Klebsiella pneumoniae, vancomycin-resistant Enterococcus (VRE), methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant (MDR) Acinetobacter baumannii, MDR Enterobacter, and combinations thereof; preferably, the target bacterium may be selected from the group consisting of extended-spectrum beta-lactamase-producing (ESBL) Escherichia coli strains.

[0111] Alternatively, the target bacteria may be bacteria of the microbiome of a given species, preferably bacteria of the human microbiota.

[0112] In one aspect, synthetic bacterial delivery vehicles with a desired target host range are provided for use in transferring a desired nucleic acid payload into target bacterial cells. The synthetic bacterial delivery vehicles may be characterized by a chimeric receptor binding protein (RBP), where the chimeric RBP comprises a fusion between the N-terminal domain of an RBP from a lambdoid bacteriophage or lambda bacteriophage and the C-terminal domain of a different RBP. It has been demonstrated that important portions of lambdoid RBPs, such as the stf protein, can be exchanged for portions of different RBPs.

[0113] As used herein, a receptor binding protein or RBP is a polypeptide that recognizes and optionally binds to and / or modifies or degrades a substrate located on a cell shell, such as, but not limited to, a bacterial outer membrane, LPS, capsule, protein receptor, channel, structure, such as a flagellum, pili, or secretion system. The substrate can be, but is not limited to, any carbohydrate or modified carbohydrate, any lipid or modified lipid, any protein or modified protein, any amino acid sequence, and any combination thereof.

[0114] The present disclosure also provides synthetic cell delivery vehicles characterized by the presence of an engineered branched receptor-binding multisubunit protein complex ("branched RBP"). Such delivery vehicles can be used to import a nucleic acid payload of interest into target bacterial cells. An engineered branched RBP comprises two or more associated receptor-binding proteins from a bacteriophage that associate with each other based on the presence of an interaction domain (ID). The association of one subunit with another can be non-covalent or covalent. Each polypeptide subunit contains an ID that serves as an "anchor" for the association of one subunit RBP with another. In certain embodiments, a branched RBP can comprise multiple RBP subunits, e.g., two, three, or four subunits.

[0115] For disclosures of recombinant bacterial delivery vehicles, see U.S. Provisional Patent Applications Nos. 62 / 849,108, 62 / 849,112, 62 / 802,777, 62 / 771,761, and 62 / 783,258, each of which is incorporated herein in its entirety.

[0116] As used herein, the term "nucleic acid payload" refers to any nucleic acid sequence or amino acid sequence, or a combination of both (e.g., but not limited to, peptide nucleic acid or peptide-oligonucleotide conjugate) transferred into a bacterium by a delivery vehicle. Preferably, the term "nucleic acid payload" refers to any nucleic acid sequence (e.g., but not limited to, peptide nucleic acid or peptide-oligonucleotide conjugate), optionally in combination with an amino acid sequence, transferred into a cell by a delivery vehicle. The nucleic acid payload is designed to encode a protein or nucleic acid of interest for transfer into a desired target bacterial host cell. The term "nucleic acid payload" can refer to a plasmid, vector, or cargo. The nucleic acid payload can be a phagemid or plasmid derived from a natural, evolved, or engineered bacteriophage genome. The nucleic acid payload can also consist of only a portion of a phagemid or plasmid derived from a natural, evolved, or engineered bacteriophage genome. The nucleic acid payload is preferably a nucleic acid sequence packaged in a bacterial delivery vehicle, preferably a bacteriophage-derived capsid.

[0117] Described herein are engineered bacterial delivery vehicles with desired target bacterial cell specificity and / or host range, each containing a nucleic acid payload with an embedded unique tracer nucleotide sequence tag. As used herein, a "multivalent mixture of delivery vehicles" refers to a mixture of one or more different or dissimilar bacterial delivery vehicles. Such bacterial delivery vehicles may differ because they have at least one structural feature that distinguishes them from other bacterial delivery vehicles in the mixture. Such structural features may include, for example, differences in bacterial cell binding ability and / or host range, which may be caused by differences in the RBPs expressed within the delivery vehicles. Thus, a multivalent mixture contains at least two distinct populations of bacterial delivery vehicles.

[0118] The pharmaceutical or veterinary compositions of the present invention are defined herein as polyvalent mixtures of bacterial delivery vehicles, i.e., containing at least two different bacterial delivery vehicles, each containing a nucleic acid payload with a unique tracer nucleic acid sequence. Pharmaceutical or veterinary compositions can be prepared as sterile solid compositions that can be suspended at the time of administration using sterile water, saline, or other appropriate sterile injectable media. Pharmaceutical or veterinary compositions of the present invention can be orally administered in the form of a sterile solution or suspension containing other solutes or suspending agents (e.g., sufficient saline or glucose to make the solution isotonic), bile salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleic acid ester of sorbitol and its anhydride copolymerized with ethylene oxide), etc. Particles according to the present invention can also be orally administered in either liquid or solid composition form. Compositions suitable for oral administration include solid forms such as pills, capsules, granules, tablets, and powders, and liquid forms such as solutions, syrups, elixirs, and suspensions. Forms useful for enteral administration include sterile solutions, emulsions, and suspensions.

[0119] The bacteriophage-derived particles of the present invention can be dissolved or suspended in a pharmaceutically acceptable liquid vehicle, such as water, an organic solvent, a mixture of both, or a pharmaceutically acceptable oil or lipid. The liquid vehicle may contain other suitable pharmaceutical additives, such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavorings, suspending agents, thickeners, colorants, viscosity adjusters, stabilizers, or osmolality adjusters. Suitable examples of liquid vehicles for oral and enteral administration include water (partially containing additives such as those described above, such as cellulose derivatives, preferably carboxymethylcellulose sodium solution), alcohols (including monohydric and polyhydric alcohols, such as glycols) and their derivatives, and oils (e.g., coconut oil and peanut oil). For parenteral administration, the vehicle can also be an oily ester, such as ethyl oleate and isopropyl myristate. Sterile liquid vehicles are useful in sterile liquid form compositions for enteral administration. The liquid vehicle for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.

[0120] For transdermal administration, the pharmaceutical or veterinary compositions may be formulated in the form of an ointment, cream, or gel, and penetration may be enhanced using suitable penetrants or surfactants, such as, for example, dimethyl sulfoxide, dimethylacetamide, and dimethylformamide.

[0121] For transmucosal administration, nasal sprays, rectal or vaginal suppositories can be used.The active compound can be incorporated into any known suppository base by methods known in the art.Examples of such bases include cocoa butter, polyethylene glycol (carbowax), polyethylene monostearate sorbitan, and mixtures thereof with other compatible materials that modify the melting point or dissolution rate.

[0122] The present invention also relates to a pharmaceutical or veterinary composition of the present invention for use in treating a disease or disorder caused by a bacterium. It also relates to a method for treating a disease or disorder caused by a bacterium, comprising administering a pharmaceutical or veterinary composition of the present invention to a subject having said disease or disorder in need of treatment. It further relates to the use of a pharmaceutical or veterinary composition of the present invention for the manufacture of a medicament for the treatment of a disease or disorder caused by a bacterium.

[0123] In one embodiment, the disease or disorder caused by bacteria is a chronic inflammation of the skin, such as acne vulgaris, abdominal cramps, acute epiglottitis, arthritis, bacteremia, bloody diarrhea, botulism, brucellosis, brain abscess, myocardial damage, chancroid, chlamydia infection, Crohn's disease, conjunctivitis, cholecystitis, colorectal cancer, polyposis, dysbiosis, Lyme disease, diarrhea, diphtheria, duodenal ulcer, endocarditis, erysipelothricosis, typhoid fever, fever, glomerulonephritis, gastroenteritis, gastric ulcer, Guillain-Barré syndrome, tetanus, gonorrhea, gingivitis, inflammatory bowel disease, irritable bowel syndrome, leptospirosis, leprosy, listeriosis, tuberculosis, Lady Windermere syndrome, syndrome, Legionnaire's disease, meningitis, mucopurulent conjunctivitis, multidrug-resistant bacterial infection, multidrug-resistant bacterial carriage, myocarditis, myonecrosis-gas gangrene, Mycobacterium avium complex, neonatal necrotizing enterocolitis, nocardiosis, hospital-acquired infection, otitis, periodontal disease, pharyngitis, pneumonia, peritonitis, purpuric fever, Rocky Mountain spotted fever, dysentery, syphilis, sinusitis, sigmoiditis, sepsis, subcutaneous abscess, tularemia, tracheobronchitis, tonsillitis, typhoid fever, ulcerative colitis, urinary infection, whooping cough, and non-alcoholic fatty liver disease (NASH).

[0124] In another embodiment, the disease or disorder caused by bacteria is preferably an infection caused by bacteria selected from the group consisting of intestinal infections, such as esophagitis, gastritis, enteritis, colitis, sigmoiditis, proctitis, and peritonitis, urinary tract infections, vaginal infections, female upper reproductive tract infections, such as salpingitis, endometritis, oophoritis, metritis, parametritis, and pelvic peritoneal infections, respiratory tract infections, such as pneumonia, intra-amniotic infections, dental infections, endodontic infections, fibrosis, meningitis, bloodstream infections, nosocomial infections, such as catheter-related infections, nosocomial pneumonia, postpartum infections, nosocomial gastroenteritis, nosocomial urinary tract infections, and combinations thereof. Preferably, the infection is caused by bacteria exhibiting antibiotic resistance. In certain embodiments, the infection is caused by a bacterium listed above as a target bacterium.

[0125] In another embodiment, the disease or disorder caused by bacteria is a metabolic disorder such as obesity and diabetes.

[0126] In certain embodiments, the present invention relates to pharmaceutical or veterinary compositions for use in treating conditions involving bacteria of the human microbiome, such as inflammatory and autoimmune diseases, cancer, infectious diseases, or brain disorders. The present invention relates to methods for treating conditions involving bacteria of the human microbiome, comprising administering a pharmaceutical or veterinary composition of the present invention to a subject having such a condition in need of treatment. The present invention also relates to the use of a pharmaceutical or veterinary composition of the present invention for the manufacture of a medicament for the treatment of conditions involving bacteria of the human microbiome. Indeed, some bacteria of the microbiome may secrete molecules that may induce and / or enhance inflammatory or autoimmune diseases or cancer development without triggering any infection. More specifically, the present invention relates to modulating the composition of the microbiome to improve the efficacy of immunotherapies based on, for example, CAR-T (chimeric antigen receptor T) cells, TILs (tumor-infiltrating lymphocytes), and Tregs (regulatory T cells), also known as suppressor T cells. Modulation of the composition of the microbiome to improve the efficacy of immunotherapy may also include the use of immune checkpoint inhibitors known in the art, such as, but not limited to, PD-1 (programmed cell death protein 1) inhibitors, PD-L1 (programmed death-ligand 1) inhibitors, and CTLA-4 (cytotoxic T-lymphocyte-associated protein 4).

[0127] Some bacteria in the microbiome can also secrete molecules that affect the brain.

[0128] Therefore, a further object of the present invention is a method for controlling the microbiome of a subject, comprising administering in said subject an effective amount of the pharmaceutical composition disclosed herein.

[0129] In certain embodiments, the present invention also relates to a personalized treatment method for an individual in need of treatment for a bacterial infection, comprising the steps of: i) obtaining a biological sample from the individual and identifying a group of bacterial DNA sequences from the sample; ii) identifying one or more pathogenic bacterial strains or species in the sample based on the sequence identification; and iii) administering to the individual a pharmaceutical composition according to the present invention capable of recognizing and delivering a packaged payload to each pathogenic bacterial strain or species identified in the sample. The present invention further relates to a) a pharmaceutical or veterinary composition of the invention for use in treating a bacterial infection in an individual, or b) use of a pharmaceutical or veterinary composition of the invention for the manufacture of a medicament in treating a bacterial infection in an individual, wherein in a) or b), preparing the pharmaceutical or veterinary composition comprises: (i) providing a biological sample from the individual and identifying a group of bacterial DNA sequences from the sample; ii) identifying one or more pathogenic bacterial strains or species in the sample based on the sequence identification; and iii) preparing a pharmaceutical or veterinary composition according to the invention that is capable of recognizing the pathogenic bacterial strains or species identified in the sample and delivering a packaged payload.

[0130] Preferably, the biological sample comprises pathogenic and non-pathogenic species, and after administration of a pharmaceutical or veterinary composition according to the invention to an individual, the amount of pathogenic bacteria on or in the individual is reduced, but the amount of non-pathogenic bacteria is not reduced.

[0131] In another particular embodiment, the present invention relates to a pharmaceutical or veterinary composition according to the present invention for use in improving the efficacy of a drug. Indeed, some bacteria of the microbiome, without being pathogenic in themselves, are known to be able to metabolize drugs and modify them into ineffective or harmful molecules.

[0132] In another specific embodiment, the present invention relates to the in situ bacterial production of any compound of interest, including therapeutic compounds, e.g., mammalian prophylactic and therapeutic vaccines. The compound of interest can be produced within the target bacterium, secreted from the target bacterium, or expressed on the surface of the target bacterium. In a more specific embodiment, an antigen is expressed on the surface of the target bacterium for prophylactic and / or therapeutic vaccination.

[0133] The present invention also relates to non-therapeutic uses of the bacterial delivery particles. For example, the non-therapeutic use may be cosmetic or for improving the health of a subject, particularly in a subject not suffering from a disease. Thus, the present invention also relates to cosmetic or non-therapeutic compositions comprising the bacteriophage-derived particles of the present invention.

[0134] Subjects, Dosage Regimen, and Administration The subject to be treated by the composition of the present invention is an animal, preferably a mammal, and even more preferably a human. However, the term "subject" can also refer to a non-human animal, particularly a mammal, such as, among others, a dog, a cat, a horse, a cow, a pig, a sheep, a donkey, a rabbit, a ferret, a gerbil, a hamster, a chinchilla, a rat, a mouse, a guinea pig, and a non-human primate, or a non-mammal, such as a poultry, in need of treatment.

[0135] The human subject may be a human in the fetal stage, a newborn, a child, an infant, an adolescent, or an adult of any age.

[0136] In a preferred embodiment, the subject has been diagnosed with or is at risk of developing an infection, disorder, and / or disease, preferably caused by a bacterium. Methods for diagnosing such infections, disorders, and / or diseases are well known to those skilled in the art.

[0137] In certain embodiments, the infection, disorder, and / or disease is caused by a bacterium that is resistant to treatment, preferably the infection, disorder, and / or disease is caused by a bacterium that is resistant to antibiotics.

[0138] In certain embodiments, the subject has not received any treatment prior to administration of a delivery vehicle according to the invention, preferably a payload according to the invention, in particular a payload packaged in a delivery vehicle according to the invention, preferably a plasmid or phagemid packaged in a bacterial viral particle according to the invention, or a pharmaceutical or veterinary composition according to the invention.

[0139] In certain embodiments, the subject has already undergone at least one series of treatments, preferably several series of treatments, prior to administration of a delivery vehicle according to the invention, preferably a payload according to the invention, in particular a payload packaged in a delivery vehicle according to the invention, preferably a plasmid or phagemid packaged in a bacterial viral particle according to the invention, or a pharmaceutical or veterinary composition according to the invention.

[0140] Preferably, the treatment is administered periodically, preferably daily to monthly, more preferably daily to every two weeks, more preferably daily to weekly, and even more preferably, the treatment is administered daily. In certain embodiments, the treatment is administered several times a day, preferably two or three times a day, and even more preferably three times a day.

[0141] The duration of treatment with a delivery vehicle according to the invention, preferably a payload according to the invention, in particular a payload packaged in a delivery vehicle according to the invention, preferably a plasmid or phagemid packaged in a bacterial viral particle according to the invention, or a pharmaceutical or veterinary composition according to the invention, is preferably comprised between 1 day and 20 weeks, more preferably between 1 day and 10 weeks, more preferably between 1 day and 4 weeks, and even more preferably between 1 day and 2 weeks. In certain embodiments, the duration of treatment is about 1 week. Alternatively, treatment may be continued for as long as the infection, disorder, and / or disease persists.

[0142] The form, route of administration and dosage of the delivery vehicle according to the invention, preferably the payload according to the invention, in particular the payload packaged in the delivery vehicle according to the invention, preferably the plasmid or phagemid packaged in the bacterial virus particle according to the invention, or the pharmaceutical or veterinary composition according to the invention can be adjusted by one skilled in the art according to the type and severity of the infection (e.g. depending on the disease, disorder, and / or bacterial species involved in the infection and its localization in the body of the patient or subject), and in particular the age, weight, sex and general physical condition of the patient or subject.

[0143] In particular a delivery vehicle according to the invention, preferably a payload according to the invention, in particular a payload packaged in a delivery vehicle according to the invention, preferably a plasmid or phagemid packaged in a bacterial virus particle according to the invention, or a pharmaceutical or veterinary composition according to the invention, to be administered. 、 The amount of should be determined by standard procedures well known to those skilled in the art. The physiological data of the patient or subject (e.g., age, size, and weight) and the route of administration should be taken into consideration to determine the appropriate dosage so that a therapeutically effective amount is administered to the patient or subject.

[0144] For example, the total amount of payload packaged in a delivery vehicle, particularly a delivery vehicle according to the invention, preferably a plasmid or phagemid packaged in a bacterial viral particle according to the invention, for each administration is 10 4 From 10 15 is included among the delivery vehicles.

[0145] As used herein, "tracer" refers to a nucleic acid sequence associated with, i.e., embedded within, a nucleic acid payload. The tracer functions as a bacterial delivery vehicle "identification tag" that can be used to assess the presence and / or quantity of the bacterial delivery vehicle in a polyvalent mixture of bacterial vehicles. As disclosed herein, the tracer can be embedded within a non-coding region, a coding region, and / or a 5' or 3' untranslated region of the nucleic acid payload.

[0146] A tracer can be any nucleic acid sequence whose detection correlates with the presence of a specific cellular delivery vehicle. Thus, in the mixture of the present invention, each population of bacterial delivery vehicles contains a different, unique tracer. Detection of the tracer sequence can be achieved using a variety of different detection methods well known to those skilled in the art. In certain embodiments, the tracer sequence can serve as a binding site for one or more primers used in conjunction with an amplification technique, such as a PCR reaction. A tracer can be designed to contain a constant region to which a primer can bind for amplification of the tracer in a mixture of bacterial delivery vehicles. Preferably, the constant region is identical in all tracers in a multivalent mixture. Such constant region-binding primers can be used to collectively amplify all tracers in a mixture of bacterial delivery vehicles. A tracer can be designed to contain a variable region to which a primer can bind for amplification of the tracer in a mixture of bacterial delivery vehicles. The variable region-binding primer can be used to identify specific bacterial delivery vehicles within a mixture. In one aspect, the variable region is located within a barcode.

[0147] To identify and / or quantify bacterial delivery vehicles in a mixture, one or more rounds of amplification can be performed. In certain embodiments, a mixture of different types of primers can be used in each amplification reaction. For example, primers that bind only to constant regions, primers that bind only to variable regions, and / or primers that overlap constant and variable regions can be used. Alternatively, in the case of an amplification reaction, primers that bind to constant regions and primers that bind to variable regions can be used. Depending on the primers used, the amplification reaction can result in the amplification of a mixture of tracers found in the mixture of bacterial delivery vehicles altogether. Alternatively, primers can be selected to amplify a specific bacterial delivery vehicle in the mixture.

[0148] In certain embodiments, primers that bind to the constant region of the tracer may be used for a first round of amplification, followed by a subsequent second round of amplification reaction, for example, using primers that bind to the variable region of the tracer, to further assess the presence of a particular delivery vehicle in a mixture of vehicles.

[0149] As described in detail below, one or more characteristics should be considered in the design and characterization of a tracer sequence for use in identifying bacterial delivery vehicles. Such characteristics include, for example, the length of the primer binding site, the melting temperature, and the percent G / C content, each of which can affect, for example, the efficiency of an amplification reaction. Furthermore, the tracer should be designed to minimize interference with the growth characteristics of the cell line packaging the bacterial delivery vehicle or the target bacterial cells. The tracer should have minimal impact on the production of the delivery vehicle in the packaging cell line. Undesirable restriction enzyme cleavage sites should be avoided. Finally, regions of homology with packaging cellular DNA, target host DNA, or payload DNA should be avoided in the tracer because such regions of homology can serve as sites for undesired homologous recombination.

[0150] In certain embodiments, a multivalent mixture of bacterial delivery vehicles (e.g., phagemid particles) containing identical nucleotide nucleic acid payloads except for a unique "tracer" nucleic acid sequence is provided, in such a way that all nucleic acid payloads are packaged in the same capsid with the same associated tracer, and nucleic acid payloads are packaged in different capsids with different associated tracers. The use of such tracers allows for the detection of the presence and / or relative abundance of each bacterial delivery vehicle in the multivalent mixture of delivery vehicles by cycles of amplification and, optionally, sequencing. Such amplification methods include, for example, PCR, qPCR, ddPCR, LCR, FISH, or NGS, using primers specific to the "tracer" sequence.

[0151] In different embodiments, the tracer nucleic acid sequence is embedded in a coding or non-coding DNA sequence, such as, but not limited to, a reporter gene (e.g., antibiotic resistance, fluorescent protein), a tag, or a barcode.

[0152] First proposed in 1982 and first put into practice in 2003, DNA barcoding suggested the existence of a single DNA locus in every species that could be used to uniquely identify each species and, through comparison and alignment, could depict a more faithful natural taxonomy than other methods (Taylor and Harris, 2012, Mol Ecol Resour. 2012 May;12(3):377-88). An emergent science on the brink of irrelevance: a review of the past 8 years of DNA barcoding. Although no single "magic bullet" barcode has been found that works for all domains of life, DNA barcodes that have been found to work exceptionally well within specific clades, such as mitochondrial cytochrome c oxidase subunit 1 (CO1) in animals or 16S rRNA in bacteria, have been widely adopted to empirically create biological taxonomy or "trees of life" and describe ecosystems (Kress et al., 2015, Trends Ecol Evol, Jan; 30(1):25-35; Taylor and Harris, 2012, Mol Ecol Resour. 2012 May; 12(3):377-88).

[0153] The advent of high-throughput DNA sequencing in 2005, and its cost since then has decreased, leading to the generalization of DNA barcoding beyond species identification. DNA barcoding is now used in a variety of contexts, including: 1) genome and chromatin mapping, 2) transcriptomics / RNA-seq, 3) ribosome profiling for analyzing protein translation activity, 4) protein engineering (via phage display, mRNA / ribosome display, and yeast two-hybrid screening), 5) DNA-encoded protein libraries, 6) cell surface DNA labeling, 7) antibody-DNA conjugates, 8) DNA-encoded small molecule and cyclic peptide libraries, and 9) nanoparticle-DNA conjugates (Liszczak and Muir, 2019, Angew Chem Int Ed Engl, Mar 22;58(13):4144–4162).

[0154] In certain non-limiting embodiments, the tracer can be designed for use in barcoding. A "barcode" represents a nucleic acid sequence whose presence correlates with the presence of its associated bacterial delivery vehicle. In such cases, the "barcode" is embedded within the tracer and is flanked on both sides by sequences (constant regions, FIG. 3) that can be utilized to amplify the barcode sequence (variable regions, FIG. 3) for identification and / or quantification of the barcode, thereby indicating the presence and quantity of a specific bacterial delivery vehicle in a polyvalent mixture of bacterial delivery vehicles. The use of such barcoding provides an easy and convenient way to use multiple unique tracers whose presence correlates with the presence of a specific bacterial delivery vehicle in a mixture of bacterial delivery vehicles.

[0155] In certain embodiments, the tracer is embedded in the nucleic acid payload within a non-coding region of DNA. As used herein, a "non-coding region of DNA" refers to a region containing a sequence that does not encode a protein. Such a tracer is introduced into the nucleic acid payload in such a manner that nucleic acid payloads with the same tracer are packaged within the same bacterial delivery vehicle. By using such a tracer nucleic acid sequence, e.g., one containing a barcode, the presence and relative abundance of each different bacterial delivery vehicle in a polyvalent bacterial delivery mixture can be confirmed by PCR, qPCR, ddPCR, LCR, FISH, or NGS using primers specific to sequences adjacent to and within the barcode.

[0156] When designing a tracer for use in the non-coding region of a nucleic acid payload, one or more factors can be considered to design an efficient and effective tracer. In one embodiment, the tracer should have minimal effect on the growth of the packaging cell line for use in producing the bacterial delivery vehicle or on the production of the bacterial delivery vehicle. Furthermore, the tracer should have minimal effect on the growth of the target bacterial cell.

[0157] Tracers can be designed in a manner that allows for the detection and quantification of theoretically any number of bacterial delivery vehicles in a mixture of said vehicles. Such detection can be achieved initially using a common site, i.e., a constant region, such as a constant region adjacent to a variable region (e.g., C1 and C2 in FIG. 2), that serves as a target for primer binding and subsequent amplification (see FIG. 2). The use of such a common site thus allows for the simultaneous co-amplification of several different tracer nucleic acid sequences. In certain embodiments, the tracer sequence may also allow for the detection of each specific tracer sequence and thus the presence of its associated bacterial delivery vehicle by use of a common primer site and / or variable primer sites per construct: V1, V2, ..., Vx (see FIG. 2).

[0158] In a preferred embodiment, the tracer can be designed to be used in conjunction with a variety of different detection methods. Such detection methods include the use of multiple amplification cycles for the detection and / or quantification of the tracer, resulting in the differentiation and / or quantification of two or more bacterial delivery vehicles in a polyvalent mixture of bacterial delivery vehicles. Such detection methods include, for example, PCR, qPCR (dye-based / probe-based), ddPCR (dye-based / probe-based), LCR, FISH, and / or NGS.

[0159] Additionally, the tracers utilized in the disclosed methods and compositions can be designed to have restricted secondary structure and optimal melting temperatures to allow binding of primers / probes to the tracer nucleic acid sequence when using PCR methods such as qPCR / ddPCR / LCR / FISH / NGS detection methods.

[0160] When considering the positioning of a tracer within a nucleic acid payload, the location should be selected to minimize transcriptional or translational interference with other functional elements of the nucleic acid payload. Ideally, the tracer should be free of undesired restriction sites, and the tracer should be designed to have limited sequence homology (>20 nt) with other regions of the nucleic acid payload, the packaging cellular DNA, or the target bacterial host cell DNA. Such homology can serve as a substrate for undesired homologous recombination events.

[0161] In certain non-limiting embodiments, a DNA barcoding system can be used for the identification of bacterial delivery vehicles (see Figure 2). A location in the DNA nucleic acid payload selected to minimize interference with its function is the ELF5 insertion site, which is the site for the linker region of standardized plasmid constructs. In such embodiments, N (N = number) random nucleic acid tracers with certain structural features are generated. In preferred embodiments, the lengths of the barcode and constant regions can typically range from 25 to 50 nucleotides each. The barcode and constant regions should be long and unique enough to allow binding of specific primers to enable PCR amplification (a necessary step for qPCR, ddPCR, and NGS methods, respectively) without generating unwanted amplicons. The selected structural features, in a non-limiting example, include a 30-nucleotide-long tracer, where the number of barcodes generated = N, where N is 1 or greater and the GC content is limited to between about 60% and 70%. The lengths of the constant region and barcode are generally determined by the melting temperature (Tm) of the desired primers used and depend on the detection method used. Each generated tracer nucleic acid sequence should be examined for the detection of undesired restriction sites. In addition, the secondary structure, melting temperature, and sequence homology can be determined to minimize secondary structure, ensure a sufficiently high melting temperature for annealing of primers targeting the tracer sequence, and at the same time minimize the possibility of homologous recombination. Methods for testing specific tracers for desirable structural features are well known to those skilled in the art. After the sequence is selected, tracer nucleic acids, which may include barcodes, are recombinantly or chemically synthesized and cloned into their respective positions within the non-coding region of the nucleic acid payload.

[0162] [Table 1]

[0163] In certain embodiments, the tracer nucleic acid sequence comprises a barcode selected from the group consisting of the b1 to b12 barcodes defined in Table 1.

[0164] In another embodiment, the tracer nucleic acid sequence can be embedded in the coding region of the nucleic acid payload, taking into account codon usage. In the case of 64 codons, three codons code for stop codons, and 61 codons code for the 20 standard amino acids. In the case of the 20 amino acids, only two (Met, Trp) are coded by one unique codon, nine (Asn, Asp, Cys, Gln, Glu, His, Lys, Phe, Tyr) are coded by two unique codons, one (Ile) is coded by three unique codons (also three unique stop codons), five (Ala, Gly, Pro, Thr, Val) are coded by four unique codons, and three (Arg, Leu, Ser) are coded by six unique codons. This degeneracy in the 18 standard amino acids (in addition to the stop codon) allows for the introduction of codon changes into a DNA sequence such that the differences introduced into the coding sequence will nevertheless encode proteins with the same amino acid sequence.

[0165] In a specific embodiment, the coding sequence with altered codon usage is introduced into the nucleic acid payload in such a way that all nucleic acid payloads packaged in the same capsid have the same tracer, and consequently, nucleic acid payloads packaged in different capsids have different tracers. In such cases, the presence and relative abundance of each cellular delivery vehicle in the multivalent mixture can be confirmed by PCR, qPCR, or ddPCR using primers specific to the tracer nucleic acid sequence.

[0166] For the design of a tracer embedded in a protein coding region, one or more structural features may be considered. Such features may include those considered in the design of a tracer inserted into a non-coding region of a nucleic acid payload. A tracer sequence inserted into a coding region should be designed to introduce codon changes into the protein coding region so that the introduced differences encode a protein with the same amino acid sequence. The tracer should have minimal impact on the efficiency of protein expression or the function of the protein.

[0167] In one aspect, the tracer should have minimal effect on the growth of the packaging bacterial cells used to produce the bacterial delivery vehicle, as well as on the growth of the target bacterial cells. Furthermore, the tracer should have minimal effect on the production of the bacterial delivery vehicle within the packaging bacterial cells.

[0168] Tracers can be designed in a manner that allows for the detection and quantification of theoretically any number of bacterial delivery vehicles in a mixture of said vehicles due to insertion into the coding region. Such detection can be achieved initially using a common site, i.e., a constant region, such as C1 and C2 in Figure 3, that serves as a target for primer binding and subsequent amplification (see Figure 3). The use of such a common site thus allows for the co-amplification of several different tracer nucleic acid sequences. In certain embodiments, the nucleic acid tracer sequence can also allow for the detection of each specific tracer nucleic acid sequence and, consequently, the presence of its homologous bacterial delivery vehicle by using one common site and variable sites per construct: V1, V2, ..., Vx (see Figure 3). Such variable sites can be utilized in an initial single amplification cycle or can be used in a second round of amplification cycles after using the common site for amplification.

[0169] Each generated tracer for insertion into the coding region can be examined for the detection of undesired restriction sites. In addition, secondary structure, melting temperature, and sequence homology are examined to minimize secondary structure and ensure a sufficiently high melting temperature for annealing of primers targeting the tracer sequence, while simultaneously minimizing the possibility of homologous recombination. 。 After the tracer sequences are selected, tracer nucleic acids, which may include barcodes, can be synthesized recombinantly or chemically and cloned into their respective locations within the non-coding regions of the DNA nucleic acid payload.

[0170] In a specific, non-limiting embodiment, a tracer system is developed as follows: A tracer sequence is selected for insertion within the coding DNA sequence (CDS) for the TEM-1 β-lactamase (bla) protein, which is commonly used to confer ampicillin resistance when expressed in cells. For the design of the tracer, the coding region is a selected (X) 10 codon in length from nucleotides 118 to 147 in the bla CDS. The tracer sequence comprises gca-cga-gtg-ggt-tac-atc-gaa-ctg-gat-ctc (SEQ ID NO: 18), which encodes the 10 amino acids ARVGYIELDL (SEQ ID NO: 19) at positions 40 to 49. Within the selected coding region (X) codons encoding the (X) amino acids (or stop codons), the methionine (M, Met) and tryptophan (W, Trp) amino acids remain unchanged because these amino acids have unique codons. In the bla example, none of the amino acids in the selected 10 amino acid segment are M or W, meaning that all have multiple codons that can code for each amino acid. For the remaining codons / amino acids in region X, a register is created for each amino acid such that the entire codon encoding that amino acid is numbered and listed (for example, Ala40: 1) GCT, 2) GCC, 3) GCA, 4) GCG; see Figure 3). Then, all combinations within each codon possibility at each of the amino acid positions in region X are identified and listed as unique strings within the complete list of all possible codon combinations.As an example, for the 10 amino acid region AA40-49 in TEM-1 β-lactamase, one possibility, and a specific number representing each of the specific possible codons in numerical form corresponding to amino acid positions 40-49, is {1, 1, 1, 1, 1, 1, 1, 1, 1, 1}; another is {2, 1, 1, 1, 1, 1, 1, 1, 1, 1}; another is {3, 1, 1, 1, 1, 1, 1, 1, 1}; etc., representing each possible codon at each amino acid position, to exhaustion of the possible unique combinations. The three exemplary representations above would correspond to three 30-nucleotide sequences in which only the first codon (GCT / GCC / GCA, 1 / 2 / 3, corresponding to the Als at position aa40) differs, whereas the subsequent 27 nucleotides would be the first possibility for each remaining amino acid (hence CGT / GTT / GGT / TAT / ATT / GAA / CTT / GAT / CTT (SEQ ID NO:20)) using the ordering of Figure 4 to represent the remaining RVGYIELDL (SEQ ID NO:21). An exhaustive comprehensive list of all possible codon combinations (or possibly only a subset to reduce computational time) is then taken, and the Hamming distance is calculated between each. In the example of TEM-1 β-lactamase, each of the 10 amino acids has multiple possible codons (A=4, R=6, V=4, G=4, Y=2, I=3, E=2, L=6, D=2, L=6), and taking these into account, the total number of codons that can be generated is the product 4*6*4*4*2*3*2*6*2*6, or 331,776.

[0171] After designing, 50 bp upstream and downstream of the 30 bp barcode, as well as the barcode itself, are examined for the presence of undesired restriction sites. Only those tracers without such restriction sites are selected for use. Furthermore, the presence of secondary structure, melting temperature, and sequence homology are examined to minimize secondary structure, ensure a sufficiently high melting temperature for the barcode-targeting primer, and minimize the possibility of homologous recombination, respectively.

[0172] The number of unique codon combinations is selected to correspond to the number of unique members of the bacterial delivery vehicle mixture, thereby selecting or favoring the maximum Hamming distance between each (Mohammadi-Kambs M. et al., 2017, ACS Omega. 2017 Apr 30; 2(4): 1302-1308) (both to enable unique identification of each member and to attempt to minimize the possibility of cross-reaction / detection among / between the unique members and unique probes / primers for each). The selected DNA barcodes are then recombinantly or chemically synthesized and cloned into their respective positions within the CDS of the DNA nucleic acid payload.

[0173] In yet another embodiment, the tracer nucleic acid sequence can be embedded in a 5' or 3' untranslated region (UTR) DNA sequence. 5' and 3' UTRs are regions at each end (5' and 3' ends, respectively) of a protein-coding sequence that are transcribed into mRNA but not translated into protein. In prokaryotic cells, 5' UTRs are usually much shorter (3-10 nucleotides in length) than 3' UTRs and have a stronger effect on protein expression than 3' UTRs. The design specifications for tracers embedded in 5' or 3' untranslated regions are the same as those used for tracers embedded in non-coding regions of nucleic acid payloads.

[0174] A method for detecting and quantifying bacterial delivery vehicles in a multivalent bacterial delivery vehicle mixture is provided, wherein each bacterial delivery vehicle in the mixture contains a nucleic acid payload with a unique tracer, and the method includes detecting and quantifying all or each of the bacterial delivery vehicles in the mixture by performing cycles of amplification using primers that bind to sequences within the tracer sequence. As described above, depending on the use of primers targeting the constant and / or variable regions of the tracer, all or only a portion of the bacterial delivery vehicles can be detected and / or quantified. This method therefore allows for the detection and / or quantification of each population of bacterial delivery vehicles contained in the multivalent mixture and / or all bacterial delivery vehicles contained in the mixture. Such a mixture of bacterial delivery vehicles containing a tracer sequence embedded in their nucleic acid payload allows for the characterization of a final pharmaceutical product by detecting and quantifying each bacterial delivery vehicle in the multivalent bacterial delivery vehicle mixture. Such methods also allow for the identification and tracking of bacterial delivery vehicles after administration to organisms, humans, and / or animal models during safety and efficacy clinical trials (excretion, biodistribution). For example, if the presence of a particular tracer sequence is found to be associated with an undesirable safety and / or biodistribution profile outcome, the homologous bacterial delivery vehicle can be removed from the bacterial delivery vehicle mixture, rendering the final drug product safe.

[0175] A rapid, high-throughput screening method for testing the activity of bacterial delivery vehicles against a collection of individual bacterial strains or a mixture of bacterial strains is provided. Such screening methods provide an efficient means for the identification of desirable bacterial delivery vehicles capable of transferring their nucleic acid payloads to specific target bacterial cells in an efficient manner.

[0176] Additionally, methods are provided for characterizing the potency and extent of sequential bacterial delivery following administration of different vehicles in the time dimension (by initial transduction with delivery vehicle 1, then allowing some time to pass, then subsequent transduction with delivery vehicle 2).

[0177] Further additional methods are provided for characterizing non-species targeting by specific bacterial delivery vehicles from a mixture when administered to a complex microbial community of two or more prokaryotic populations, including, but not limited to, a human microbiome sample.

[0178] Also provided is a method for detecting and tracking bacterial delivery vehicles after administering a multivalent mixture of bacterial delivery vehicles to an organism, each bacterial delivery vehicle comprising a nucleic acid payload with a unique tracer, and detecting and quantifying each of the bacterial delivery vehicles in the mixture by performing cycles of amplification, and optionally sequencing, using primers that bind to sequences within the tracer. Such amplification methods include, for example, PCR, qPCR, ddPCR, or NGS. [Example]

[0179] Example 1 Plasmid pAK272B (SEQ ID NO: 1) was modified to contain one of seven different barcodes (SEQ ID NOs: 2 to 8; B1, B2, B3, B4, B5, B6, and B7 in Figure 5) to yield seven distinguishable plasmid variants designated pB1, pB2, pB3, pB4, pB5, pB6, and pB7. DNA concentrations were assessed for each plasmid variant based on absorbance at 260 nm and used to create equimolar mixtures with a goal of having 1250 copies of each plasmid variant per microliter of mixture. The absolute copy numbers of each variant in the mixture were then experimentally confirmed by droplet digital PCR (ddPCR). To achieve this, 20 microliters of ddPCR reactions were designed with 100 nM of a primer that binds to a common region of the pAK272B plasmid backbone (SEQ ID NO: 9; referred to as C1 in Figure 5), 100 nM of another primer that binds specifically to a single selected barcode (SEQ ID NOs: 10 to 16; referred to as V1, V2, V3, V4, V5, V6, and V7 in Figure 5), 8 microliters of barcoded plasmid mixture (i.e., 500 copies of each variant per microliter of final ddPCR reaction), and a commercially available ddPCR master mix (QX200 ddPCR EvaGreen Supermix) containing polymerase enzyme, dNTPs, and DNA dye. A total of seven reactions were set up, plus seven negative controls in which the plasmid mixture was replaced with water, one reaction for each primer pair specific to an individual barcode. The ddPCR workflow was performed as recommended by the manufacturer. As shown in Figure 6, the number of each barcoded variant was measured within the expected order of magnitude (227 copies per microliter versus an expected approximately 500 copies per microliter, i.e., a difference of 0.3 log), with the small difference likely due to the absorbance method used to estimate the initial DNA concentration. Despite this, similar copy numbers were found for all seven plasmid variants, as originally intended.

[0180] Example 2 Four different barcoded variants of the plasmid pAK272B were used to identify differences in in vivo delivery rates to target bacteria among the four lambda phage-derived scaffolds. Each barcoded plasmid was packaged within one lambda phage-derived scaffold variant and delivered at 2 x 10 per microliter of each. 7 By mixing the particles, all four mixtures resulting in packaged phagemids were obtained. 100 microliters of this mixture was administered to nine mice colonized with the target strain MG-GFP, and for each animal, 10-fold serial dilutions (from pure to 10) of resuspended feces were administered. -5 Successfully transduced bacteria were recovered from mouse feces by plating 4 μl of the plasmid pAK272B (up to 4 μl) onto selective medium containing chloramphenicol (plasmid pAK272B carries a chloramphenicol resistance gene). The entire transduced bacterial population contained within the 4 μl patch of pure fecal sample was then grown in vitro in the presence of chloramphenicol, and any plasmids present were purified using standard plasmid miniprep techniques. Total DNA concentration was estimated using absorbance at 260 nm, and a sufficient amount of DNA was subjected to ddPCR, aiming for >80,000 copies per reaction, assuming all purified and detected DNA corresponds to barcoded plasmids. One reaction was set up with a specific primer pair for each barcode, resulting in a total of four reactions per original mouse sample. Figure 7 shows the absolute number of each barcoded plasmid recovered from the transduced bacteria for each mouse in the experiment (M10 to M18). The clear abundance of barcode B4 across all nine animals indicates that the lambda phage-derived scaffold variant packaged with that specific plasmid variant was most effective in delivering to target bacteria in vivo. Other plasmid variants were also detected but at lower copy numbers, indicating that the corresponding lambda phage-derived scaffolds were delivered less efficiently in vivo.

Claims

1. A polyvalent mixture of bacterial delivery vehicles comprising at least two different bacterial delivery vehicles which are bacteriophage-derived scaffolds and differ in bacterial cell binding ability and / or host range, wherein each bacterial delivery vehicle comprises a phagemid having an embedded unique tracer, the phagemids have identical sequences except for the tracer, and the tracer comprises a barcode that is between 25 and 50 nucleic acids in length.

2. 10. The multivalent mixture of bacterial delivery vehicles of claim 1, wherein the tracer is embedded in a non-coding region.

3. 10. The multivalent mixture of bacterial delivery vehicles of claim 1, wherein the tracer is embedded in the coding region.

4. A method for detecting and / or quantifying bacterial delivery vehicles in a polyvalent bacterial delivery vehicle mixture comprising at least two different bacterial delivery vehicles which are bacteriophage-derived scaffolds and differ in bacterial cell binding ability and / or host range, wherein each bacterial delivery vehicle comprises a phagemid having a unique tracer, the method comprising detecting and / or quantifying (i) each bacterial delivery vehicle and / or (ii) all of the bacterial delivery vehicles by amplification of the tracer using primers which bind to the unique tracer sequence, the phagemids having identical sequences except for the tracer, and the tracer comprising a barcode which is between 25 and 50 nucleic acids in length.

5. A method for detecting and / or quantifying bacterial delivery vehicles after administering to a subject a polyvalent bacterial delivery vehicle mixture comprising at least two different bacterial delivery vehicles that are bacteriophage-derived scaffolds and differ in bacterial cell binding ability and / or host range, wherein each bacterial delivery vehicle comprises a phagemid having a unique tracer nucleic acid sequence, the method comprising detecting and / or quantifying (i) each bacterial delivery vehicle and / or (ii) all of the bacterial delivery vehicles in a sample derived from the subject by performing multiple cycles of amplification using primers that bind to the unique tracer sequences, the phagemids having identical sequences except for the tracer, and the tracer comprising a barcode that is between 25 and 50 nucleic acids in length.

6. The method of claim 4 or 5, wherein the tracer is embedded in a non-coding region.

7. 6. The method of claim 4 or 5, wherein the tracer is embedded in the coding region.

8. 8. The method of any one of claims 4 to 7, wherein detection and quantification of each bacterial delivery vehicle is carried out by performing multiple cycles of amplification using primers that bind to unique tracer nucleic acid sequences.

9. A pharmaceutical composition comprising a polyvalent mixture of bacterial delivery vehicles comprising at least two different bacterial delivery vehicles which are bacteriophage-derived scaffolds and differ in bacterial cell binding ability and / or host range, wherein each bacterial delivery vehicle comprises a phagemid having a unique tracer nucleic acid sequence, the phagemids have identical sequences except for the tracer, and the tracer comprises a barcode which is between 25 and 50 nucleic acids in length.

10. 10. The pharmaceutical composition of claim 9 for use in the treatment of a disease or disorder caused by bacteria.

11. 11. The pharmaceutical composition of claim 10, wherein the disease or disorder caused by bacteria is selected from the group consisting of infectious diseases caused by bacteria, metabolic disorders, and pathologies involving bacteria of the human microbiome.

12. The pharmaceutical composition described in claim 11, wherein the metabolic disorder is obesity or diabetes.

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