Novel compositions of matter comprising proteins that incorporate PFS230 or PVS230 domain 1 (PFS230d1 or PVS230d1) at the n-terminus, malaria vaccine antigens, and their use

Abstract

The invention provides a protein comprising, from amino- to carboxy-termini: (a) a first polypeptide domain derived from the Pfs230 domain 1 (Pfs230D1) of Plasmodium falciparum or the Pvs230 domain 1 (Pvs230D1) of Plasmodium vivax, (b) an optional linker domain, and (c) a second polypeptide domain, which can be a 6-Cys polypeptide, wherein the fusion protein is not the native Pfs230 or Pvs230 protein. The invention also provides nucleic acids encoding the inventive protein, genetic vectors comprising such nucleic acid, cells comprising the protein, nucleic acid or vector, compositions comprising such reagents. The invention also provides a method for vaccinating a subject using the inventive protein and composition and the use of the inventive protein and composition in connection with such a method.

Description

NOVEL COMPOSITIONS OF MATTER COMPRISING PROTEINS THAT INCORPORATE PFS230 OR PVS230 DOMAIN 1 (PFS230D1 OR PVS230D1) AT THE N-TERMINUS, MALARIA VACCINE ANTIGENS, AND THEIR USECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This patent application claims the benefit of U.S. Provisional Patent Application No. 63 / 617,717 filed January 4, 2024, which is incorporated by reference.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with Government support under project number AI001007- 14 by the National Institutes of Health, National Institutes of Health (NIH) National Institute of Allergy and Infectious Diseases. The Government has certain rights in the invention.INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY

[0003] Incorporated by reference in its entirety herein is a computer-readable nucleotide / amino acid sequence listing submitted concurrently herewith and identified as follows: One 152,650 Byte XML file named “772344. XML,” dated January 3, 2025.BACKGROUND OF THE INVENTION

[0004] Malaria transmission-blocking vaccines reduce disease transmission by breaking the continuous cycle of infection between the human host and the mosquito vector. Malaria-exposed human populations have been observed to acquire antibodies against Pfs230, a parasite protein expressed by gametocytes in the human stage of Plasmodium falciparum and a surface antigen of gametes and zygotes in the mosquito stage. Hence, Pfs230 has emerged as a target for designing transmission-blocking vaccines.

[0005] The DI domain of Pfs230 (Pfs230Dl, SEQ ID NO: 1) is the N-terminal domain of Pfs230 and is a 6-cystein-containing (“6-Cys”) polypeptide, as are other domains of Pfs230. Duein part to ease of production and functional activity in phase 1 and 2 clinical trials, Pfs230Dl has emerged as a leading transmission-blocking vaccine candidate. In addition Pfs48 / 45, which is another 6-Cys protein from P. falciparum, shows potential functional activity as a transmission blocking candidate. However, it would be desirable to add to the vaccine arsenal represented by these two polypeptides. Other domains from Pfs230 may appear to be logical choices for investigation; however, likely due, in part, to their also being 6-Cys polypeptides, expression of them has proven to be technically challenging. The present invention addresses these technical concerns.BRIEF SUMMARY OF THE INVENTION

[0006] In one aspect, the invention provides a protein comprising, from amino- to carboxytermini: (a) a first polypeptide domain derived from the Pfs230 domain 1 (Pfs230Dl) of P. falciparum or the Pvs230 domain 1 (Pfs230Dl) of P. vivax, (b) an optional linker domain, and (c) a second polypeptide domain, which can be a 6-Cys polypeptide, wherein the inventive protein is not the full native Pfs230 protein.

[0007] The invention also provides nucleic acids encoding the inventive protein, genetic vectors comprising such nucleic acid, cells comprising the protein, nucleic acid or vector, compositions comprising such reagents. The invention also provides a method for vaccinating a subject using the inventive protein and composition and the use of the inventive protein and composition in connection with such a method.BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0008] Figure 1 presents schematic illustrations of the organization of the polypeptide domains within the Pfs230 (top) and Pfs48 / 45 proteins.

[0009] Figure 2 presents Western blot data concerning glycosylated (G) and nonglycosylated (NG) versions of Pfsl30Dl and Pfsl30Dl-D3. The indicated samples were probed with a functional human monoclonal antibody recognizing Pfs230Dl.

[0010] Figure 3 presents data concerning the antibody titers at 21, 42, and 56 days postinjection in mice treated with either Pfsl30Dl (bottom panel) or Pfsl30Dl-D3 (top panel). Data for Green Fluorescent Protein (GFP) are presented as a control.

[0011] Figure 4 presents Western blot data concerning the activity of proteins, including fusion protein Pfs230Dl-Pfs48D3, probed either with a conformationally-sensitive antibody specific for either Pfs48 / 45 (left panel) or Pfs230Dl (right panel).

[0012] Figure 5 presents data comparing the serum activity of rabbits that had been inoculated either with 25 pg or 50 pg Pfs230Dl-Pfs48D3 or with 50 pg PDF230D1 (either as the form used with the proteins described here (PDF230D1G) or as the monomeric form used in LMIV transmission blocking vaccine (PDF230D1M).

[0013] Figure 6 presents images of Coomassie stained gels representing expression experiments for Pfsl30D7, Pfsl30Dl / D7, Pfsl30D9, and Pfsl30Dl / D9. The rightmost images reflects similar data for purified Pfsl30Dl / D9.

[0014] Figure 7 graphically presents data concerning the functional activity of antibodies raised against Pfs230D2, Pfs230D3, and PFS230D9 from rabbits inoculated with either Pfs230Dl-D2, Pfs230Dl-D3, or Pfs230Dl / D9.

[0015] Figure 8 presents a plot of data concerning the oocyst-reducing activity (TRA) of serum from rabbits inoculated with either Pfs230Dl, Pfs230Dl-D3, or Pfs230Dl / D9. The data are reflective of the activity of serum diluted at 1 :8, 1 : 16, 1 :32, or 1 :64, as indicated.

[0016] Figure 9 presents Western blot data concerning the expression of Pfs230Dl fusions with either Pfs230 / D9 or Pfs45 / 48D3, wherein the two domains had been separated by linkers of two, four, six, or eight peptide linkers (one (xl), two (x2), three (x3), or four (x4) repeats of the dipeptide “TG,” respectively). These also included a His tag for purification. The blots were probed with an antibody recognizing the His tag. The top panel presents an image of the blot, whereas the bottom panel presents the quantitative values of the expression of the monomeric band of each, relative to the data for the respective xl species of each.

[0017] Figure 10 presents Western blot data concerning the expression of Pfs45 / 48, wherein the two domains had been separated by linkers of two, four, six, or eight peptide linkers (one (xl), two (x2), three (x3), or four (x4) repeats of the dipeptide “TG,” respectively). The blotswere probed with a conformation-dependent functional monoclonal antibody against Pfs230Dl (antibody 3E12). The top panel presents an image of the blot, whereas the bottom panel presents the quantitative values of the expression of the monomeric band relative to the data for the respective xl species (left panel) or normalized to the expression data for His discussed with respect to Figure 9.

[0018] Figure 11 presents Western blot data concerning the expression of either Pfs230Dl- Pfs48D3 or Pvs230Dl-Pfs48D3. The blots were probed with an antibody recognizing a His tag.

[0019] Figure 12 presents sequence comparison between orthologous 230D1 domains of Plasmodium falciparum and P. vivax (portions of SEQ ID NOs: 1 and 36).

[0020] Figure 13 presents images of SDS-PAGE of small-scale purification (left) of Pvs230Dl / D8 fusion at 2 different amounts loaded on the gel and right) SDS-PAGE of scaled up after 2 column purification. Both gels samples were loaded non-reduced (nr) or reduced (r).

[0021] Figure 14 presents an image of a Western blot of spent media from small scale transfections using an anti-his antibody for detection. PfCSP was included as a positive transfection control.

[0022] Figure 15 presents data from an ELISA assay demonstrating that Pfs230Dl antibodies were removed from the total IgG of animals immunized with either Pfs230Dl-3 (top) or Pfs230Dl / D9 (bottom). Both Pfs230Dl (left) and the entire antigen (right) were used as the plate antigen.

[0023] Figure 16 presents data representing standard membrane feeding assay (“SMFA”) activity comparing antibodies specific to Pfs230Dl or Pfs230D12-3 run at 2 concentrations.

[0024] Figure 17 presents data concerning functional ELISA pools of human monoclonal antibodies at increasing amounts of non-functional antibodies, demonstrating non-functional antibodies are blocked in the assay (top panel). Functional ELISA measurement at 4 different dilutions of pooled rabbit serum from the 4 different constructs comparing the overall functional response (bottom panel).

[0025] Figure 18 presents data concerning the transmission-reducing activity from 2 biological replicates of affinity purified Pfs230Dl from pooled IgG of rabbits immunized with different Pfs230 constructs as indicated.

[0026] Figure 19 presents data concerning the expression of Pfs230Dl / Pfs230D14. The top image depicts a Western blot showing that Pfs230D14 is not expressed alone but in a fusion monomeric protein is observed. The lower image depicts SDS-PAGE following a two-step purification of Pfs230Dl / Pfs230D14, demonstrating that the monomeric fusion is recoverable.DETAILED DESCRIPTION OF THE INVENTIONInventive Protein

[0027] In one aspect, the invention provides a protein comprising, from amino- to carboxy termini: (a) a first polypeptide domain derived from the Pfs230 domain 1 (Pfs230Dl) of P. falciparum, (b) an optional linker domain, and (c) a second polypeptide domain, wherein the inventive protein is not the native Pfs230 protein (i.e., not the full-length native Pfs230 protein). Similarly, the inventive protein can comprise, from amino- to carboxy termini: (a) a first polypeptide domain derived from the Pvs230 domain 1 (Pvs230Dl) of P. vivax, (b) an optional linker domain, and (c) a second polypeptide domain, wherein the inventive protein is not the native Pvs230 protein (i.e., not the full-length native Pvs230 protein).

[0028] As noted, Pfs230Dl, as a stand-alone protein is undergoing clinical investigation, and it is known in the art. Its sequence is set forth herein as SEQ ID NO: 1. In the context of the inventive protein, the first polypeptide domain derived from Pfs230Dl can comprise an amino acid sequence consisting of SEQ ID NO: 1, or its amino acid sequence can consist essentially of SEQ ID NO: 1, or the amino acid sequence of the first polypeptide domain can comprise SEQ ID NO: 1. Similarly, in the context of the inventive protein, the first polypeptide domain derived from Pvs230Dl can comprise an amino acid sequence consisting of SEQ ID NO:36, or its amino acid sequence can consist essentially of SEQ ID NO:36, or the amino acid sequence of the first polypeptide domain can comprise SEQ ID NO:36.

[0029] It will be observed that, so long as the first polypeptide domain retains the capacity to act as a chaperone domain, enhancing the expression or production of the C-terminal second domain, it can be incorporated into the inventive protein. Thus, for example, the first polypeptide domain can comprise a nucleic acid sequence that differs from that of SEQ ID NO: 1 or SEQ ID NO:36, such as being at least being at least 80% identical, or at least 80% identical, orat least 90% identical, or at least 95% identical, or at least 99% identical to SEQ ID NO: 1 or SEQ ID NO:36. As mentioned, in some embodiments, the first polypeptide domain can consist of SEQ ID NO:1 or SEQ ID NO:36, i.e., being 100% identical to either.

[0030] Furthermore, the sequence of amino acids of the first polypeptide domain can comprise substations relative to SEQ ID NO: 1 or SEQ ID NO:36, which can be “conservative” but need not be. In this respect, “conservative” substitutions involve, as non-limiting examples, (a) substituting an acid amino acid (e.g., Aspartate, Glutamate, Asparagine, Glutamine) with another acid amino acid, (b) substituting an aliphatic amino acid (e g., Glycine, Alanine, Valine, Leucine, or Isoleucine) with another aliphatic amino acid, (c) substituting an aromatic amino acid (e.g., Phenylalanine, Tyrosine, or Tryptophan) with another aromatic amino acid, (d) substituting a hydroxyl or sulfur / selenium-containing amino acid (e.g., Serine, Cysteine, Selenocysteine, Threonine, or Methionine) with another sulfur / selenium-containing amino acid, or (e) substituting a basic amino acid (e.g., Histidine, Lysine, or Arginine) with another basic amino acid.

[0031] Additionally, the sequence of amino acids of the first polypeptide domain can include N-terminal additions of between 1 and 10 amino acids. In this respect, proteins produced and tested as reported in the Examples below contain a first polypeptide domain comprising SEQ ID NO: 1 or SEQ ID NO:36 with an N-terminal addition of the dipeptide “TG.”

[0032] Within the inventive protein, when present, the optional linker domain is a polypeptide comprising between 0 or 1 and 30 amino acids, such as between 1 and 20 amino acids, between 1 and 15 amino acids, or between 1 and 10 amino acids, between 2 and 8 amino acids having been tested and found active in the Examples below. In the exemplary sequences, one or more tandem repeats of the amino acids, “TG” serve as linker domains, when present. However, the identity of the amino acids for the linker domain can vary from these. Of course, an embodiment of the inventive protein need not contain a linker domain, in which instance the first polypeptide domain derived from the Pfs230 domain 1 (Pfs230Dl) of P. falciparum or derived from the Pvs230 domain 1 (Pvs230Dl) of P. vivax is fused, as an N-terminal domain, to the second polypeptide domain. Where the inventive protein comprises a linker domain,preferably the amino acids comprising the linker do not form a secondary structure, such as an a- helix or a P-sheet.

[0033] Within the inventive protein, the second polypeptide domain can comprise any desired polypeptide, so long as the inventive protein does not represent the native, full-length, Pfs230 or Pvs230 protein. In certain embodiments, the second polypeptide domain within the inventive protein can be derived from or comprise, consist essentially of, or consist of a 6-Cys polypeptide, which can be derived from the Pfs230 protein (such as, but not limited to, Domain 2 (Pfs230D2, e.g., SEQ ID NO:14), Domain 3 (Pfs230D3, e.g., SEQ ID NO:15), Domain 4 (Pfs230D4), Domain 7 (Pfs230D7, e.g., SEQ ID NOs: 10-13), Domain 9 (Pfs230D9, e.g., SEQ ID NOs:6-9), Domain 14 (Pfv230D14, e.g., SEQ ID NO:80), etc., thereof). However, the second polypeptide domain within the inventive protein can alternatively or in addition be derived from or comprise, consist essentially of, or consist of other polypeptides, including those from other proteins from P. falciparum for example, as set forth as SEQ ID NOs: 2-5 herein (e.g., Pfs48 / 45D3) and discussed below in Examples 3 and 6. Some exemplary amino acid sequences from which the second polypeptide domains for inclusion inventive protein can be derived from or comprise, consist essentially of, or consist of are set forth herein as SEQ ID NOS: 1, 36, 38, 42, 46, 50, 54, 58, 62, 70, 78, 84, 86, 88, 90, and 92, which are provided as illustrative examples and not for the purpose of limiting the scope of the invention to such sequences.

[0034] Moreover, the second polypeptide domain within the inventive protein can be derived from or comprise, consist essentially of, or consist of polypeptides from species other than P. falciparum. For example, the second polypeptide within the Pfs230Dl / Pvs48D3 lx TG embodiment presented herein (SEQ ID NO: 16) and discussed below in Example 7 is derived from P. vivax. Similarly, SEQ ID NO: 40 (Pfs230Dl / Pvs230D2 IxTG) represents a Pfs23Dl- fusion, wherein the second polypeptide domain is derived from P. vivax Pvs230D2; SEQ ID NO: 44 (Pfs230Dl / Pvs230D3 IxTG) represents a Pfs23Dl -fusion wherein the second polypeptide domain is derived from P. vivax Pvs230D3; SEQ ID NO: 48 (Pfs230Dl / Pvs230D4 IxTG) represents a Pfs23Dl -fusion wherein the second polypeptide domain is derived from P. vivax Pvs230D4; SEQ ID NO: 52 (Pfs230Dl / Pvs230D5 IxTG) represents a Pfs23Dl -fusion wherein the second polypeptide domain is derived from P. vivax Pvs230D5; SEQ ID NO: 56(Pfs230Dl / Pvs230D2-3 IxTG) represents a Pfs23Dl -fusion wherein the second polypeptide domain is derived from P. vivax Pvs230D2-3 in tandem; SEQ ID NO: 60 (Pfs230Dl / Pvs230D13-14 IxTG) represents a Pfs23Dl -fusion wherein the second polypeptide domain is derived from P. vivax Pvs230D13-14 in tandem; SEQ ID NOs: 64 (Pfs230Dl / Pvs230D7 IxTG) and 93 (Pfs230Dl / Pvs230D7 4xTG) represent Pfs23Dl -fusions wherein the second polypeptide domain is derived from P. vivax Pvs230D7; SEQ ID NO: 71 (Pfs230Dl / Pvs230D8 IxTG) and 96 (Pfs230Dl / Pvs230D8 4xTG) represent Pfs23Dl -fusions wherein the second polypeptide domain is derived from P. vivax Pvs230D8; SEQ ID NOs: 99 (Pfs230Dl / Pvs230Dl IxTG) and 100 (Pfs230Dl / Pvs230Dl 4xTG) represent Pfs23Dl -fusions wherein the second polypeptide domain is derived from P. vivax Pvs230Dl. Indeed, the second polypeptide domain within the inventive protein can be derived from or comprise, consist essentially of, or consist of proteins or polypeptides from a polypeptide derived from a species within the aconoidasidan (hematozoan) clade of Apicocomplexa, which includes both hemosporidiuma (Plasmodium) and piroplasms (see, e.g., PUB MED: 11163248, PUBMED: 16155126, PUBMED: 20386715, PUBMED: 22493233, and PUBMED:23511632).

[0035] To assist in constructing proteins according to the invention in which the second polypeptide domain within the inventive protein can be derived from or comprise, consist essentially of, or consist of proteins or polypeptides from Pfs230 or Pfs48 / 45, these proteins are schematically represented, with an identification of their domains, in Figure 1. Moreover, amino acid sequences of embodiments in which exemplary sequences thereof are derived from Pfs230, Pvs230, Pfs48 / 45, or Pvs48D3 are presented herein as SEQ ID NOs:2-18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100, which are provided as illustrative examples and not for the purpose of limiting the scope of the invention to such sequences. Given the data presented in Example 7 herein, SEQ ID NO: 17 is not preferred. Also, it will be noted that the amino acid sequences of these embodiments contain an N-terminal “TG” dipeptide, which is optional, as discussed above.

[0036] While SEQ ID NOs:2-18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93- 100 are provided as exemplary amino acid sequences representing those specific embodiments of the inventive protein tested as reported in the Examples below, as the second polypeptide domainwithin the inventive protein can be derived from or comprise, consist essentially of, or consist of polypeptides from species other than P. falciparum or from other domains of Pfs230, the exact sequence of an embodiment of the inventive protein can vary from those presented in SEQ ID NOs:2-18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100. Moreover, as discussed above with respect to the first domain, the inventive protein can have an amino acid sequence comprising minor variants of SEQ ID NOs:2-18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100. Thus, the inventive protein can comprise a nucleic acid sequence that differs from that of SEQ ID NOs:2-18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100, such as being at least being at least 80% identical, or at least 80% identical, or at least 90% identical, or at least 95% identical, or at least 99% identical to SEQ ID NOs:2-18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100. Furthermore, the sequence of amino acids of the inventive protein can comprise substations relative to SEQ ID NOs:2-18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100, which can be “conservative” but need not be conservative.

[0037] The inventive protein can additionally comprise other elements. In this respect, for example, the inventive protein can be part of a fusion sequence or larger sequence comprising the inventive protein or be conjugated with other agents. In embodiments, such larger sequence can include fusion partners or conjugation with agents for enhancing immunogenicity, such as EPA or CRM (see, for example, Healy et al., J Clin Invest. 202I;I31(7):el46221, doi: 10.1172 / JCI146221, Shimp et cz / ., Vaccine. 2013 Jun 19;31(28):2954-62. doi:10.1016 / j. vaccine.2013.04.034. epub 2013 Apr 24 (reporting the development of a Pfs25-EPA malaria transmission blocking vaccine as a chemically conjugated nanoparticle), Scaria et al., Vaccine 38(34), 5480-5489 (2020) (presenting a comparison of carrier proteins to conjugate malaria transmission blocking vaccine antigens, Pfs25 and Pfs230), and Miura et al., Vaccine, 37(15), 2073-2078 (2019), the contents of each of which are incorporated herein in their entireties).

[0038] The inventive protein also can be presented on particles (e g., microparticles or Virus- Like Particles (VLPs)), or formulated with liposomes. In this respect, conjugation can be prepared by standard methods known to those of ordinary skill in the art. For example, particlescan be formed by modifying the inventive protein with bifunctional linkers that covalently attach the antigen to the carrier. Alternatively, the inventive protein can be expressed in frame with a polypeptide that will form a self-assembled nano-particle. Suitable other approaches have been reported, which can be adapted for use with the inventive protein (see, e.g., Salinas el al., NPJ Vaccines. 2023 Aug 18;8(1): 124. doi: 10.1038 / s41541-023-00709-8 (reporting an antigen designed from a single-component 60-copy Pfs230Dl nanoparticle); Huang et al., Commun Biol. 2022 Aug 1;5(1):773. doi: 10.1038 / s42003-022-03688-z.PMID: 3591522 (reporting vaccine codisplay of CSP and Pfs230 on liposomes targeting two P. falciparum differentiation stages); and Yenkoi di ok-Douti et al., Sci Rep. 2019 Nov 14;9(1): 16833. doi: 10.1038 / s41598-019-53208- z.PMID: 31727945 (describing a VLP as an antigenic platform for a Pfs47-targeted malaria transmission-blocking vaccine), the contents of each of which are incorporated herein in their entireties).

[0039] The inventive protein can be prepared by expressing a nucleic acid encoding an appropriate amino acid sequence for the inventive protein in a cell and purifying the inventive protein from the cell (e.g., by gel electrophoresis or chromatography). Desirably, the cell employed for such expression is from an insect (wherein baculoviral expression systems are appropriately used), a mammalian cells (e.g., Chinese hamster ovary (CHO) cells, HEK293 cells, and the like) or a yeast cell (e.g., Pichia pastoris Where, as noted above, the inventive protein is part of a fusion sequence or larger sequence, or includes a conjugate partner, such can be made by expressing a nucleic acid encoding the inventive protein and its larger portion as noted. Conjugation, such as with EPA, CRM, and the like, can be accomplished by methods known to those of ordinary skill, such as chemical conjugation.Nucleic Acids

[0040] The invention also provides a nucleic acid molecule comprising a nucleic acid sequence encoding the inventive protein. The inventive nucleic acid molecule can comprise DNA (typically cDNA) or RNA and can be single or double stranded. Furthermore, the inventive nucleic acid molecule can comprise nucleotide analogues or derivatives (e.g., inosine or phosphonothioate nucleotides and the like). The inventive nucleic acid sequence can encodethe inventive protein alone or as part of a fusion sequence or larger sequence comprising the inventive protein. Persons of ordinary skill are able to deduce the coding sequence of a nucleic acid when given the sequence of the encoded inventive protein. By way of non-limiting example, nucleic acids encoding the proteins set forth in SEQ ID NOs:2-18 are set forth below as SEQ ID Nos: 19-35, and additional nucleic acids encoding other constructs are set forth below as SEQ ID NOs: 39, 43, 47, 51, 55, 59, 63, 65, 67, 72, 73, 75, 79, and 81.

[0041] The inventive nucleic acid encoding the inventive protein can be provided as part of a construct comprising the inventive nucleic acid molecule and elements that enable delivery of the nucleic acid molecule to a cell, and / or expression of the nucleic acid molecule in a cell. Such elements include, for example, expression vectors, promoters, and transcription and / or translation control sequences. Such constructs can also be referred to as "recombinant nucleic acid molecules."

[0042] The inventive nucleic acid can be made by any suitable method, such as solid-state synthesis, known to persons or ordinary skill.

[0043] Although the phrases “nucleic acid” and "nucleic acid molecule" primarily refer to the physical nucleic acid molecule and the phrase "nucleic acid sequence" primarily refers to the sequence of nucleotides on the nucleic acid, the two phrases can be used interchangeably, especially with respect to a nucleic acid molecule, or a nucleic acid sequence, being capable of encoding the inventive protein. Similarly, the phrase "recombinant nucleic acid molecule" primarily refers to a nucleic acid molecule operatively linked to an element such as a transcription control sequence but can be used interchangeably with the phrase "nucleic acid molecule."Vectors

[0044] The invention further provides a vector comprising the inventive nucleic acid sequence encoding the inventive protein. Examples of suitable vectors include plasmids (e.g., DNA plasmids), bacteria, yeast, listeria, and viral vectors, including poxvirus, retrovirus, adenovirus, adeno-associated virus, herpes simplex virus, polio virus, alphavirus, baculovirus,Sindbis virus, and the like. Where the vector is a plasmid, the plasmid can be complexed with one or more agents for facilitating transfection of cells or enhancing stability, e.g., chitosan.

[0045] The inventive vector can include suitable promoters and regulatory elements, such as one or more transcriptional regulatory element or an enhancer, which can be operably linked to the nucleic acid molecule encoding the inventive protein or other coding sequences. Suitable promoters include the SV40 early promoter, an RSV promoter, the retrovirus LTR, the adenovirus major late promoter, the human CMV immediate early I promoter, and various poxvirus promoters, such as the Pr7.5K promoter, 30K promoter, 40K promoter, 13 promoter, Prs promoter, PrsSynllm promoter, PrLEl promoter, synthetic early / late (sE / L) promoter, HH promoter, 1 IK promoter, and Pi promoter. While the promoters typically will be constitutive promoters, inducible promoters also can be used in the inventive vectors. Such inducible systems allow regulation of gene expression.

[0046] In addition to the nucleic acid molecule encoding the inventive protein and suitable regulatory elements, a vector useful in the invention (e.g., a plasmid or a viral vector) also can comprise nucleic acids encoding other proteins, such as those conferring antibiotic resistance to efficiently screen for transformants. In certain embodiments, the vector also can comprise a nucleic acid sequence encoding one or more immunostimulatory / regulatory molecules, granulocyte macrophage colony stimulating factor (GM-CSF), cytokines, and / or molecules that can enhance an immune response (e.g., additional tumor-associated antigens). Exemplary additional tumor-associated antigens (TAAs, also referred to as cancer antigens) include, but are not limited to, 5-a-reductase, a-fetoprotein (AFP), AM-1, APC, April, B melanoma antigen gene (BAGE), P-catenin, Bcll2, bcr-abl, Brachyury, CA-125, caspase-8 (CASP-8 also known as FLICE), Cathepsins, CD19, CD20, CD21 / complement receptor 2 (CR2), CD22 / BL-CAM, CD23 / FcsRII, CD33, CD35 / complement receptor 1 (CR1), CD44 / PGP-1, CD45 / leucocyte common antigen (LCA), CD46 / membrane cofactor protein (MCP), CD52 / CAMPATH-1, CD55 / decay accelerating factor (DAF), CD59 / protectin, CDC27, CDK4, carcinoembryonic antigen (CEA), c-myc, cyclooxygenase-2 (cox-2), deleted in colorectal cancer gene (DCC), DcR3, E6 / E7, CGFR, EMBP, Dna78, famesyl transferase, fibroblast growth factor-8a (FGF8a), fibroblast growth factor-8b (FGF8b), FLK-l / KDR, folic acid receptor, G250, G melanomaantigen gene family (GAGE-family), gastrin 17, gastrin-releasing hormone, ganglioside 2 (GD2) / ganglioside 3 (GD3) / ganglioside-monosialic acid-2 (GM2), gonadotropin releasing hormone (GnRH), UDP-GlcNAc:RiMan(al-6)R.2 [GlcNAc to Man(al-6)] p 1 ,6-7V- acetylglucosaminyltransferase V (GnT V), GP1, gp 100 / Pme 117, gp-100-in4, gpl5, gp75 / tyrosine-related protein- 1 (gp75 / TRP-l), human chorionic gonadotropin (hCG), heparanase, Her2 / neu, human mammary tumor virus (HMTV), 70 KD heat-shock protein (HSP70), human telomerase reverse transcriptase (hTERT), insulin-like growth factor receptor- 1 (IGFR-1), interleukin- 13 receptor (IL-13R), inducible nitric oxide synthase (iNOS), Ki67, KIAA0205, K-ras, H-ras, N-ras, KSA, LKLR-FUT, melanoma antigen-encoding family (MAGE-family, including at least MAGE-1, MAGE-2, MAGE-3, and MAGE-4), mammaglobin, MAP17, Mel an- A / mel anoma antigen recognized by T-cells-1 (MART-1), mesothelin, MIC A / B, MT-MMPs, mucin (e.g., MUC1), testes-specific antigen NY-ESO-1, osteonectin, pl5, P170 / MDR1, p53, p97 / melanotransferrin, PAI-1, platelet-derived growth factor (PDGF), pPA, PRAME, probasin, progenipoietin, prostate-specific antigen (PSA), prostate-specific membrane antigen (PSMA), RAGE-1, Rb, RCAS1, mutated Ras, SART-1, SSX-family, STAT3, STn, TAG-72, transforming growth factor-alpha (TGF-a), transforming growth factor-beta (TGF-P), Thymosin-beta-15, tumor necrosis factor-alpha (TNF-a), TP1, TRP-2, tyrosinase, vascular endothelial growth factor (VEGF), ZAG, pl6INK4, and glutathione-S-transferase (GST), as well as modified versions thereof (e.g., CEA-6D).

[0047] The vector can be constructed by any suitable method, typically by cloning the desired elements into a vector backbone. In the case of a viral vector, the nucleic acid encoding the inventive protein, as well as any other exogenous gene(s) or regulatory element(s), preferably is / are inserted into a site or region (insertion region) in the vector that does not affect the viability of the resultant recombinant virus. Such regions can be readily identified by testing segments of virus DNA for regions that allow recombinant formation without seriously affecting virus viability of the recombinant virus and are generally known to those of skill in the art. For example, a thymidine kinase (TK) gene, such as is present in many viruses, can serve as an insertion region.

[0048] The vector can be used in the production of the inventive protein by transforming cells, which then can express the inventive protein. Also, the vector can, in some embodiments, be used in therapeutic applications or for research.Cells

[0049] In one aspect of the invention, a cell (e.g., isolated cell) comprising (1) an inventive protein, (2) an inventive nucleic acid sequence encoding an inventive protein, and / or (3) a vector comprising an inventive nucleic acid molecule also is provided herein. Typically, the cell is a mammalian cell, such as are typically employed for the production of recombinant proteins. For example, HEK293 cells were employed to produce the fusion protein embodiments in the Examples below. However, a person of ordinary skill in the art can select a suitable cell for transformation with the nucleic acid sequence encoding an inventive protein (see, e.g., Schutz et al., STAR Protocols 4(4), 102572 (2023), which is incorporated herein in its entirety), such as Chinese hamster ovary (CHO) cells, which are often employed for production of biopharmaceuticals. Of course, the cell type can be non-mammalian but desirably is eukaryotic, such as insect cells employed in a baculovirus transformation system, yeast, etc.

[0050] The inventive cell can be made using standard techniques, which are known to those of ordinary skill. In general, a source cell (or population of cells) is obtained and then can be transduced with a vector (i.e., comprising an open reading frame (ORF) that encodes the inventive protein) to introduce the genetic construct into the cell (or population of cells). Thereafter, the cell is cultured under conditions suitable for expression of the ORF to produce the inventive protein within the cell. Typically, the protocol also involves culturing the cell to proliferate it into a population of cells. The population then can be purified, if desired, and then transduced with a vector, such as described herein, encoding the inventive protein.

[0051] Whichever protocol is employed, the inventive cell of course can be proliferated to generate a population of like cells (or a heterogenous population comprising the inventive cell). Such a population of cells can include, for example, at least 1 C cells / ml, such as at least 106cells / ml, such as at least 107cells / ml, such as at least 109cells / ml, or even greater densities, if desired. It will be observed that the inventive cell (or population thereof) can be used inproduction of the inventive protein, in immunological research in vitro or, in some applications, therapeutically.Compositions

[0052] The inventive protein, nucleic acid, vector, or cell can be formulated as a composition (e.g., pharmaceutical preparation) comprising the inventive protein, nucleic acid, vector, or cell, and a carrier (e.g., a pharmaceutically or physiologically acceptable carrier). Furthermore, the inventive protein, nucleic acid, vector, cell, or composition of the invention can be used in the methods described herein alone or as part of a pharmaceutical formulation.

[0053] The composition (e.g., pharmaceutical preparation) can comprise more than one inventive protein, nucleic acid, vector, or cell of the invention. Vectors and compositions of the invention can further include or can be administered with (concurrently, sequentially, or intermittently with) any other agents or compositions or protocols that are useful for inhibiting, preventing, or treating a disease or clinical condition. For example, the composition can comprise one or more other pharmaceutically active agents or drugs. Examples of such other pharmaceutically active agents or drugs that may be suitable for use in the pharmaceutical composition include, but are not limited to, EPA or CRM, and the like, which, in certain embodiments, the inventive protein can be conjugated to.

[0054] The inventive composition additionally can comprise one or more immunostimulatory / regulatory molecules or adjuvants. Any suitable immunostimulatory / regulatory molecule or adjuvant can be used, such as interleukin (IL)-2, IL- 4, IL-6, IL- 12, IL- 15, IL-15 / IL-15Ra, IL-15 / IL-15Ra-Fc, interferon (fFNj-y, tumor necrosis factor (TNF)-a, B7.1, B7.2, ICAM-1, ICAM-2, LFA-1, LFA-2, LFA-3, CD70, CD-72, RANTES, G-CSF, GM-CSF, OX-40L, 41 BBL, anti-CTLA-4, IDO inhibitor, anti-PDLl, anti- PD1, and combinations thereof. In one embodiment, the IL-12 is NHS-IL12, which is an immunocytokine composed of two IL-12 heterodimers fused to the NHS76 antibody (see Strauss et al., Clinical Cancer Research, 25(1): 99-109 (2019)). In an embodiment, the composition can include a combination of B7.1, ICAM-1, and LFA-3 (also referred to as TRICOM). The one or more immunostimulatory / regulatory molecules can be administered in the form of a vector (e.g.,a recombinant viral vector, such as those discussed herein or otherwise known to those of skill in the art) comprising a nucleic acid encoding one or more immunostimulatory / regulatory molecules. For example, the one or more immunostimulatory / regulatory molecules (e.g., IL-12) can be administered in the form of a DNA plasmid with or without chitosan. Alternatively, the one or more immunostimulatory / regulatory molecules or adjuvants can be administered as a protein (e.g., recombinant protein), such as a protein (e.g., recombinant IL-12) with or without being admixed with chitosan. Other suitable adjuvants for use within the inventive composition include alum, such as ALHYDROGEL® adjuvant 2%, ADIUVANT SYSTEM AS01 (“AS01”), a liposome-based vaccine adjuvant system containing two immunostimulants: 3-O-desacyl-4'- monophosphoryl lipid A (MPL) the saponin, QS-21 (see Didierlaurent etal., Expert Rev. Vaccines, 16(1), 55-63 (2017), the contents of which are incorporated herein in their entirety), and MATRIX M (manufactured by NOVAVAX), an adjuvant comprising saponins from Quillaja saponaria.

[0055] The carrier used in the inventive composition can be any of those conventionally used and is limited only by physio-chemical considerations, such as solubility and lack of reactivity with the active compound(s), and by the route of administration. The pharmaceutically acceptable carriers described herein, for example, vehicles, adjuvants, excipients, and diluents, are well-known to those skilled in the art and are readily available to the public. It is preferred that the pharmaceutically acceptable carrier be one which is chemically inert to the active agent(s) and one which has no detrimental side effects or toxicity under the conditions of use.

[0056] The choice of carrier and manner of formulation of the inventive composition will be determined in part by the particular cell, or composition thereof of the invention and other active agents or drugs used, as well as by the particular method used to administer the inventive composition. A variety of suitable formulations of the pharmaceutical composition, thus, can be employed in the inventive compositions, and in carrying out the inventive methods described herein. The following formulations for parenteral, subcutaneous, intravenous, intramuscular, and intraperitoneal administration are exemplary and are in no way limiting. One skilled in the art will appreciate that these routes of administering the inventive protein, nucleic acids, vectors, cells, and compositions of the invention are known, and, although more than one route can beused to administer a particular compound, a particular route can provide a more immediate and more effective response than another route.

[0057] Injectable formulations are among those formulations that are preferred in accordance with the present invention. The requirements for effective pharmaceutical carriers for injectable compositions are well-known to those of ordinary skill in the art (see, e.g., Pharmaceutics and Pharmacy Practice, J.B. Lippincott Company, Philadelphia, PA, Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., pages 622- 630 (1986)).

[0058] Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The inventive protein, nucleic acid, vector, cell, and composition can be administered in a physiologically acceptable diluent in a pharmaceutical carrier, such as a sterile liquid or mixture of liquids, including water, saline, aqueous dextrose and related sugar solutions, an alcohol, such as ethanol, isopropanol, or hexadecyl alcohol, glycols, such as propylene glycol or polyethylene glycol, dimethylsulfoxide, glycerol ketals, such as 2,2-dimethyl-l,3-dioxolane-4-methanol, ethers, such as polyethylene glycol) 400, an oil, a fatty acid, a fatty acid ester or glyceride, or an acetylated fatty acid glyceride with or without the addition of a pharmaceutically acceptable surfactant, such as a soap or a detergent, suspending agent, such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

[0059] Oils, which can be used in parenteral formulations, include petroleum, animal, vegetable, and synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, petrolatum, and mineral. Suitable fatty acids for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

[0060] Suitable soaps for use in parenteral formulations include fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (a) cationic detergentssuch as, for example, dimethyl dialkyl ammonium halides, and alkyl pyridinium halides, (b) anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylenepolypropylene copolymers, (d) amphoteric detergents such as, for example, alkyl-b-aminopropionates, and 2-alkyl-imidazoline quaternary ammonium salts, and (e) mixtures thereof.

[0061] Preservatives and buffers may be used. In order to minimize or eliminate irritation at the site of injection, such compositions may contain one or more nonionic surfactants having a hydrophile-lipophile balance (HLB) of from about 12 to about 17. The quantity of surfactant in such formulations will typically range from about 5% to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol.

[0062] The parenteral formulations can be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient, for example, water, for injections, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets.Uses

[0063] Using the inventive reagents (i.e., the inventive protein, nucleic acid, vector, cell, and composition) the invention provides the clinical application of such in methods for vaccinating a subject. In accordance with the inventive method and use, the inventive protein, nucleic acid, vector, cell, and / or composition is administered to the subject in an immunogenically-effective amount and manner of delivery suitable to elicit an immune response in the subject to the inventive protein. Thereafter, the subject acquires immunity to a pathogen (e.g., a parasite) comprising a source protein from which the inventive protein used in the method is derived.

[0064] The subject can be any animal (typically a mammal), which is in need of vaccination. Thus, the subject can be, for example, animals of veterinary importance (e.g., cats, dogs, horses,etc.), agricultural importance (e.g., cattle, sheep, goats, fowl, and the like), zoological importance (e.g., amphibians, reptiles, birds, mammals, such as induvial members of threatened or endangered species), laboratory specimens (e.g., mice, rats, etc.), and primates, including humans. In this respect, the inventive method and use can be employed in the context of veterinary or medical vaccination involving human or animal patients.

[0065] The dosage of the inventive protein to be administered in the context of the present invention can be any suitable dosage to achieve therapeutic effectiveness. The suitable dosage will be determined by the treating physician or veterinarian in the exercise of professional judgment. However, for human patients, exemplary suitable dosages of the inventive protein can fall in a range of dosages from about 1 pg to about 100 pg, with dosages from 5 pg to 50 pg being desirable (e.g., such as 5 pg, 10 pg, 15 pg, 20 pg, 25 pg, 30 pg, 35 pg, 40 pg, 45 pg, or 50 hg)-

[0066] As a result of vaccination with the inventive reagent, the subject acquires immunity to a pathogen (e.g., a parasite) comprising a protein from which the inventive protein used in the method is derived. As noted, the second polypeptide within the inventive protein can be derived from any desirable species, such as those within the aconoidasidan (hematozoan) clade of Apicocomplexa. The specific exemplary proteins employed in the Examples below incorporated polypeptides from either P. falciparum or P vivax, which are vector-bom parasites causing malaria. Accordingly, in some embodiments, the inventive method and use involving the inventive protein and compositions can be used as a transmission-blocking vaccine against malaria.EXAMPLES

[0067] The following experimental working Examples further illustrate the invention but, of course, should not be construed as in any way limiting its scope. In brief, the experiments discussed in the Examples demonstrate the production and testing of several proteins according to the invention.EXAMPLE 1

[0068] This example demonstrates the construction of a protein according to the invention and that that antibodies exhibiting high functional activity against it are elicited in antiserum from immunized animals.

[0069] Two truncated proteins were designed: Pfs230Dl-D3 (SEQ ID NO: 15) and Pfs230Dl-D4 (SEQ ID NO: 18). In these proteins, the Pfs230Dl (SEQ ID NO: 1) domain is linked with portions of the Pfs230 protein through domain 3 (“Pfs230D3”) or Pfs230 domain 4 (“Pfs230D4”). Two versions of each were constructed; one in which the native N-glycosylation sites were mutated and the other with the wild-type N-glycosylation sites intact. These were tested for expression in HEK293 cells, the results of which study revealed that the two variants having mutated N-glycosylation sites did not express in HEK293 cells, but those with wild-type sequences did express in HEK293. Western blot data concerning Pfs230Dl-D3 (glycosylated and non-glycosylated) and Pfs230Dl are depicted in Figure 2, with a functional anti-Pfs230Dl human monoclonal antibody as the probe.

[0070] These results support the hypothesis that Pfs230Dl facilitates expression of downstream 6-cys proteins. Furthermore, the data reveal that the Pfs230Dl-D3 protein is recognized by functional anti-Pfs230Dl human monoclonal antibodies.EXAMPLE 2

[0071] This example demonstrates the results of in vivo animal studies involving Pfs230Dl- D3.

[0072] Rabbits and mice were injected with either Pfs230Dl or Pfs230Dl-D3. At days 21, 42, and 56 post-injection, serum was collected an assayed for activity. The results are presented in Figure 3. Day 56 serum also was tested by a standard membrane-feeding assay (“SMFA”). Two feed cohorts were studied, and each exhibited higher oocyst-reducing activity (%TRA) for Pfs230Dl-D3 than for Pfs230Dl at day 56 (cohort 1 : 68% TRA (Pfs230Dl) vs 84% TRA (Pfs230Dl-D3 cohort 2: 62% TRA (Pfs230Dl) vs 92% TRA (Pfs230Dl-D3).

[0073] The results of these studies further support the hypothesis that Pfs230Dl facilitates expression of downstream 6-cys proteins. Furthermore, the data reveal that Pfs230Dl-D3 exhibits comparable-to-superior antigenicity than Pfs230Dl alone.EXAMPLE 3

[0074] This example demonstrates the construction of a fusion protein according to the invention and that that antibodies exhibiting high functional activity against it are elicited in antiserum from immunized animals.

[0075] A fusion protein in which Pfs230Dl (SEQ ID NO: 1) is an N-Terminal fusion partner with the to the Pfs48 / 45 domain 3 (Pfs48 / 45D3), separated by a dipeptide linker “TG,” was constructed (SEQ ID NO:2, “Pfs230Dl / Pfs48D3 lx TG”). This fusion protein was investigated in experiments in comparison with unfused Pfs48 / 45D3 alone and with the unfused Pfs230Dl (SEQ ID NO: 1) alone.

[0076] Firstly, the fusion protein, Pfs230Dl / Pfs48D3 lx TG, reacted with conformationdependent functional monoclonal antibodies against both Pfs230Dl (antibody 3E12) and Pfs48 / 45D3 (antibody LMIV230-01), which demonstrates properly folded proteins. In contrast, unfused Pfs48 / 45D3 was poorly expressed alone and did not form the conformation-dependent epitopes, demonstrating incorrect folding. These data are presented in Figure 4.

[0077] To assess the functionality of the Pfs48 / 45D3 in the fusion protein, small animal vaccination studies were conducted. Both the Pfs230Dl (including SEQ ID NO: 1 (also referred to as “Pfs230Dl G”) and a monomeric version, “Pfs230Dl M” and Pfs230Dl / Pfs48D3 lx TG were compared in inoculated rabbits. Day 56 serum was pooled for each group and tested by SMFA. All groups exhibited high TRA activity (Figure 5) at relatively high serum concentrations (1 :8 dilution). Also, since the highly active Pfs230Dl was included in all these immunizations, higher dilutions are needed to see any enhancement in the TRA due to the Pfs48 / 45D3 portion. The 50 pg dose of the Pfs230Dl / Pfs48D3 lx TG fusion retained >80% activity after 1 :32 dilution and >50% activity after 1 :64 dilution. This is in stark contrast to Pfs230Dl, which lost significant activity when the dilution exceeded 1 : 16. Also, a 25 pg dose of the Pfs230Dl / Pfs48D3 lx TG fusion displayed comparable activity to 50 pg Pfs230Dl alone,despite having half the amount of the Pfs230Dl in the vaccine; thus, the Pfs230Dl / Pfs48D3 lx TG fusion displayed higher activity than Pfs230Dl on equivalent molar basis. Additional experiments demonstrated that IgG containing antibodies raised against Pfs48 / 45D3 had functional activity.

[0078] The results of these studies further support the hypothesis that Pfs230Dl facilitates expression of downstream proteins. Taken together, the data also suggest that the Pfs230Dl / Pfs48D3 lx TG fusion protein is more immunogenic than the Pfs230Dl alone.EXAMPLE 4

[0079] This example demonstrates the construction of additional fusion proteins according to the invention.

[0080] Two additional fusion proteins were designed: Pfs230Dl / D9 lx TG (SEQ ID NO:6) and Pfs230Dl / D7 lx TG (SEQ ID NO: 10). In these fusion proteins, the Pfs230Dl domain is an N-Terminal fusion partner with either the Pfs230 domain 9 (“Pfs230D9”) or domain 7 (“Pfs230D7”), the two being separated by a dipeptide linker “TG.” The expression of these fusion constructs was compared to the expression of the unfused Pfs230D7 and Pfs230D9 polypeptides alone (i.e., not fused with the PfsDl domain).

[0081] First, small-scale expression preparations of either unfused Pfs230D7 and Pfs230D9 alone and the fusion proteins (Pfs230Dl / D7 lx TG and Pfs230Dl / D9 lx TG) were prepared. The results (Figure 6) reveal that unfused Pfs230D7 and Pfs230D9 expression was not observed. However, expression of both Pfs230Dl / D7 lx TG and Pfs230Dl / D9 lx TG was observed. Furthermore, Figure 6 (right column) also reveals Pfs230Dl / D9 lx TG was further purified to milligram quantities.

[0082] The results of these studies further support the hypothesis that Pfs230Dl facilitates expression of downstream 6-cys proteins.EXAMPLE 5

[0083] This example demonstrates that antibodies against Pfs230Dl-2 and Pfs230Dl-3 have functional activity.

[0084] To assess the antigenic activity of Pfs230Dl-D2 (SEQ ID NO: 14), Pfs230Dl-D3 (SEQ ID NO: 15), and Pfs230Dl / D9 lx TG (SEQ ID NO:6), the ability of antibodies against the Pfs30D2, D3, and D9 domains to reduce oocysts (TRA) was assessed after affinity-depleting Pfs2310D1 antibodies from serum of immunized rabbits. In essence, to evaluate the activity of antibodies targeting domains other than Pfs230Dl (i.e., Pfs30D2, D3, and D9), the antibodies targeting such domains were removed from the antiserum of rabbits that received the truncated or fusion protein. To do so Pfs230Dl was covalently conjugated to chromatography resin to generate an affinity column that specifically bound antibodies raised against domain 1 (Pfs230Dl). Rabbit antiserum was flowed over the column, which bound the Pfs230Dl antibodies, leaving antibodies which specifically bound the other domains (depleted). Functional activity was compared to the intact portion containing Pfs230Dl antibodies (intact). The results are presented in Figures 7 and 8 and in Table 1 (see especially columns under “Depleted”).Table 1

[0085] The results reveal that at least antibodies raised against Pfs230Dl -D2 and Pfs230Dl- D3 have functional oocyst-reducing activity. The results for antibodies raised against Pfs230Dl / D9 lx TG were inconclusive; they exhibited minimal TRA, but they were used at low titers.EXAMPLE 6

[0086] This example demonstrates the construction of additional fusion proteins according to the invention with varying lengths of linkers.

[0087] Additional fusion proteins were designed, modifying fusion proteins Pfs230Dl / Pfs48D3 lx TG and Pfs230Dl / D9 lx TG, to include 2, 3, and 4 “TG” repeats in the linker separating the N-terminal Pfs230Dl domain and either the Pfs48D3 or the Pfs230D9 domains. These sequences are presented herein as SEQ ID NOs: 2-5 (representing the Pfs230Dl / Pfs48D3 fusions having linkers of 1, 2, 3, and 4 TG repeats, respectively) and SEQ ID NOs: 6-9 (representing the Pfs230Dl / D9 fusions having linkers of 1, 2, 3, and 4 TG repeats, respectively).

[0088] These eight fusion constructs were expressed in HEK293 cells, and the cell supernatants probed via Western blot using an anti-His antibody; the expression levels then were quantified with respect to levels of the monomeric band. The results, depicted in Figure 9, reveal that increasing the linker link from two amino acids (“TG”) to four or more amino acids more than doubled the level of expression of the Pfs230D9 fusions, whereas the increase in expression of the Pfs48D3 fusions exhibited a more modest increase in expression correlating with the length of the linker.

[0089] The four Pfs230Dl / Pfs48D3 fusions (lx TG, 2x TG, 3x TG, and 4x TG) also were probed via Western blot using the 3E12 antibody, the expression levels then were quantified with respect to levels of the TGxl version and also normalized to the expression levels from the anti- His Western blot assessment reported above. The results, depicted in Figure 10, reveal that each of the four fusions of Pfs230Dl and Pfs48D3 was functional and that increasing the linker link from two amino acids (“TG”) to four or more amino acids resulted in a modest increase in 3E12 reactivity.EXAMPLE 7

[0090] This example demonstrates the specificity of the chaperone activity attributed to the Pfs230Dl domain in the context of the proteins according to the present invention.

[0091] Heretofore, the proteins discussed in these Examples have employed the DI domain of the Pfs230 protein, which is derived from P. falciparum, as the first domain (N-terminal domain). The results presented in Examples 1-6 reveal that Pfs230Dl possesses a remarkable ability to serve as a chaperone when N-terminally fused to second domains, boosting expression of such second domains. To explore the specificity of this chaperone activity, additional fusion proteins were constructed in which the D3 domain of Pvs230, derived from derived from P. vivax, replaced the orthologous PfsD3 domain of the fusion proteins discussed above. One of these fusion proteins (Pfs230Dl / Pvs48D3 lx TG) employed the P. falciparum Pfs230Dl as the N-terminal first domain, as has been the case for the fusion proteins discussed in Examples 1-6; its sequence is presented as SEQ ID NO: 16. The other of these fusion proteins (Pvs230Dl / Pvs48D3 lx TG) also replaced the Pfs230Dl N-terminal first domain with the orthologous Pvs230Dl domain (SEQ ID NO:36) from P. vivax; the amino acid sequence of Pvs230Dl / Pvs48D3 lx TG is presented as SEQ ID NO: 17.

[0092] These fusion constructs were expressed in HEK293 cells, and the extracts probed via Western blot using an anti-His antibody. The results, depicted in Figure 11, reveal that Pfs230Dl / Pvs48D3 lx TG was expressed as a monomeric protein, but that Pvs230Dl / Pvs48D3 lx TG was not expressed.

[0093] One conclusion flowing from these data is that the chaperone effect of the P. falciparum 230D1 domain may be specific, as the orthologous domain of the related P. vivax species exhibited no similar effect. A sequence comparison of the two 230D1 domain orthologs (Pfs vs. Pvs) is presented in Figure 12, which reveals that the two domains are 46% identical and 63% similar. To investigate this, additional fusion proteins were designed in which the N- terminal (first) domain is derived from Pvs230Dl domain the C-terminal (second) domain was derived from Pvs230D7 or Pvs230D8, each with either 1 or 4 “TG” spacers. These sequences are presented herein as SEQ ID NOs: 66 and 68 (representing the Pvs230Dl / Pvs230D7 fusions having linkers of 2 and 4 TG repeats, respectively), SEQ ID NOs: 74 and 76 (representing thePvs230Dl / Pvs230D8 fusions having linkers of 2 and 4 TG repeats, respectively). Both domains expressed well as could be seen by Coomassie stained gel of the transfection supernatant (Figure 13). Pvs230Dl / Pvs230D8 with a 4x linker was scaled up, generating 6.93 mg from a 250 mL culture (27.7 mg / L). Taken together these results suggest that Pvs230Dl could have chaperone activity ofPfs230Dl.

[0094] Further investigation focused on whether other Pvs230 domains could be expressed as fusions with Pfs230D 1. Genetic fusions were made in which the N-terminal (first) domain is derived from the Pfs230Dl domain, upstream to either Pvs230D2 (SEQ ID NO:40), Pvs230D3 (SEQ ID NO: 44), Pvs230D4 (SEQ ID NO:48), Pvs230D5 (SEQ ID NO:52), Pvs230D7 (SEQ ID NOs:64), Pvs230D8 (SEQ ID NO:71), Pvs230D2-3 (SEQ ID NO:56), and Pvs230D13-14 (SEQ ID NO:60). High levels of expression were observed for Pvs230D7 and D8 while some expression was seen for D2, D3 and D4 (Figure 14). Generating a fusion of Pfs230Dl and a 6C double domain at the C-terminus did not permit expression of the fusion protein, suggesting that a single 6C domain can be chaperoned by Pfs230Dl but not a tandem of 2 domains. . Together this result demonstrates that in addition to Pvs48D3, other 6C domains from Pvs230 can be expressed as fusions with Pfs230Dl.EXAMPLE 8

[0095] This example explores the enhanced activity of Pfs230 fragments and fusions from a mechanistic standpoint.

[0096] There are 3 potential mechanisms of enhanced activity. First, functional antibodies directed against the other domains (i.e. Pfs230D2, D3, D9, Pfs48D3) could contribute antibodies that have are functional and fix complement mediated lysis of the parasite or inhibit other biological functions of Pfs230Dl. Antibodies against other targets could synergize with antibodies against Pfs230Dl and have improved and more durable activity. Second, having other domains downstream of Pfs230Dl could improve the quality of the Pfs230Dl functional response. Since a portion of the Pfs230Dl vaccine is occluded in the native structure, antibodies raised against this occluded face cannot bind native Pfs230 and will provide no functional activity. A response to this non-productive face could compete with antibodies that could providefunctional activity and dimmish the overall functional response and we can improve the overall functional response by focusing it to the functional face. Third, the total levels of Pfs230Dl antibodies could be improved. The additional domains could include helper T-cell epitopes that could improve the total response to the antigen increasing the total levels of Pfs230Dl antibodies (functional and non-functional or just functional). This Example investigates these possibilities.

[0097] Since it is not possible to generate antibodies against other Pfs230 domains such as D2, D2-3 or D9 without Pfs230Dl present, a purification and depletion strategy was used to fractionate antibodies raised in rabbits against the different antigens (Pfs230Dl-2, Pfs230Dl-3, Pfs230Dl / D9). Using affinity resin coupled with Pfs230Dl alone, IgG can be separated into 2 specificities: Pfs230Dl alone or the other domains in the construct and test the novel specificity for functional activity. The following workflow was employed to do this.

[0098] First, total IgG was purified from a pool of 5 animals immunized with either Pfs230Dl-3 or Pfs230D9. Next an antigen specific column was made by covalently cross-linking Pfs230Dl-3 or Pfs230Dl / D9 to a 5 mL Hi-Trap NHS-Ester column. IgG was run over the column to purify the total antigen IgG against the immunized antigen. Next, a Pfs230Dl specific column was made by covalently crosslinking Pfs230Dl to a 5 mL Hi-Trap NHS-Ester column. The total antigen (Pfs230Dl-3 or Pfs230Dl / D9) was run over the Pfs230Dl specific column. The remaining antibodies in the flow through represented antibodies specific to Pfs230D2-3 or Pfs230D9. As verified via ELISA (Figure 15), all the Pfs230Dl antibodies were removed from the sample, and the only specificity remaining in the flow-through was against the entire antigen and not domain 1. The total response of Pfs230Dl / D9 was majority Pfs230Dl, and insufficient antibodies specific to Pfs230D9 were recovered to measure functional activity. At these low levels, much contribution to activity is unlikely, and the increased activity must be another mechanism.

[0099] Next functional activity was measured by the standard membrane feeding assay (SMFA) to compare the antibodies of Pfs230D2-3 vs. the Pfs230Dl antibodies (Figure 16). Two concentrations were compared in the assay and run with 2 biological replicates. SMFA on the Pfs230Dl fraction had 100% transmission-reducing activity compared to the Pfs230D2-3antibodies, which had high transmission reducing activity at 200 pg / mL. This suggests that antibodies against Pfs230D2-3 have some functional activity but not as much as Pfs230Dl.

[0100] Next, the functional response of Pfs230Dl antiserum immunized with the different Pfs230D fusions and fragments was examined. A functional ELISA that measures only antibodies bound to the functional face of Pfs230Dl and blocks antibodies generated against the occluded face was employed. Blocking was achieved by generating a biotinylated antibody fragment called a FAB, which could bind antigen but lacked the Fc portion of the antibody. The FAB recognized the non-functional face of Pfs230Dl. The non-functional FAB was coated to the ELISA plate, and the Pfs230Dl antigen was bound to the FAB. Binding to the FAB blocked the non-functional face but allowed functional face antibodies to bind (Figure 17, top). Therefore, serum from vaccinated rabbits will only see functional antibodies bind. The functional response of Pfs230Dl-2, Pfs230Dl-3, Pfs230Dl / D9 and Pfs230Dl alone were compared. Pooled rabbit serum was tested at 4 different dilutions (Figure 17, bottom). Comparing the 1 : 80,000 and 1 :320,000 dilutions the Pfs230Dl-3 had the highest functional titer, followed by D1 / D9 and then DI -2 and last DIG. These data suggest that extending the domain increases the functional response and could contribute to the improved Pfs230Dl responses.

[0101] Next, whether the total amount of Pfs230Dl antibodies raised against our construct varied by construct and if their activity differed based on the construct was examined. This experiment was designed to permit monitoring of the yields and extract a serum mg / mL amount of Pfs230Dl specific IgG. Therefore, rabbit IgG was purified from pooled serum, and it was noted how much serum was used to generate the IgG (the total IgG per mL of rabbit serum being known). The serum was purified twice of the IgG column to verify all the IgG was removed. Next, a Pfs230Dl specific affinity column was used, applying a total of 10 mg of total IgG from each group. Pfs230Dl-3 had the highest amount of DI specific antibodies, followed by Pfs230Dl / D9 the Pfs230DlG and curiously Pfs230Dl-2 had the worst response (Table 2).Table 2Yields of Pfs230Dl specific IgG and amount present in serum from 2 biological replicates.Pfe230D1-2 42.5 52 4.3% 5.2% 0.12 0.136Pfs230D1-3 102.5 96 10.2% 9.6% 0.24 0.234Pfs230D1 / D9 87.5 83 8.8% 8.3% 0.20 0.192Pfe230D1G 90 86 9.0% 8.7% 0.18 0.165

[0102] Next, the function and binding affinity of the of the Pfs230Dl antibodies isolated from the different fusions and fragments was tested. First the SMFA was conducted at 3 different concentrations of Pfs230Dl specific antibodies. Pfs230Dl-3 had the best overall activity with TRA >60 measured at the 25 ug / mL concentration (Figure 18). Pfs230Dl alone, and Pfs230Dl / D9 had similar levels of activity at each of the dilutions, suggesting that Pfs230Dl-3 has slightly more functional antibodies then the others. Curiously, Pfs230Dl-2 has the worse activity than Pfs230Dl alone despite having the exact sequence. It is possible that Pfs230Dl-2 is a worse antigen because it is missing some of the structure that Pfs230Dl-3 seems to rescue and make the overall response better.

[0103] Improved Pfs230Dl activity could be modulated by how well antibodies bind to Pfs230Dl. Therefore, the binding affinity to Pfs230Dl of Pfs20Dl -specific antibodies was measured by Octet (Table 3). The Kd, association and dissociation rates were obtained by fitting biding curves to a 1 :1 binding model.Table 3Binding affinity of Pfs230Dl specific antibodies to Pfs230Dl as measured by Octet BLI.Kd (nM) Kd error Chi-squared R squared ka (1 / ms) ka error kd (1 / ms) kd errorPfs230Dl-2 5.7 0.017 0.0956 0.9999 75010 132.4 0.4257 0.001069Pfs230Dl-3 2.9 0.008 0.2025 0.9999 144000 145.8 0.001057 0.001057Pfs230Dl / D9 2.7 0.007 0.183 0.9999 134900 126.8 0.4257 0.000937Pfs230DlG 3.4 0.009 0.1211 0.9999 114500 124.7 0.3915 0.00095

[0104] Biotinylated Pfs230Dl was immobilized to the pin and Pfs230Dl specific antibodies from each group was measured. The Kd, Konand Koir were determined to be quite similar for all antibodies. These results suggest that affinity against the antigen did not change, but the quantity of functional antibodies could explain the difference in activity. Curiously, the Pfs230Dl-2 antibodies had the worse binding by about 2-fold. This suggests that a structural defect in Pfs230Dl-2 could have impaired a good Pfs230Dl-2 response and, therefore, that slightly lower affinity antibodies could have been raised, contributing to reduced function.

[0105] An additional fusion (Pfs230Dl / Pfs230D14) also has been tested for expression. Expression data concerning Pfs230Dl / Pfs230D14 is depicted in Figure 19. The results reveal that Pfs230D14 expression was not observed when expressed alone (upper panel), but monomeric protein was present when expressed as the fusion construct. Furthermore, following a two-step purification, the monomeric protein was recoverable (lower panel).BIOLOGICAL SEQUENCES

[0106] The following sequences are referenced herein. Underlining denotes the sequences comprising the optional linker.

[0107] Pfs230Dl Amino Acid Sequence (SEQ ID NO:1)SVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLK PTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNR GIVEVYVEPYGNK1NG

[0108] Pfs230Dl / Pfs48D3 lx TG Amino Acid Sequence (SEQ ID NO:2)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGEKKVIHGCNFSSNVSSKHTFTDSLDISLVDDSAHISCNVHLSEPKYNHLVGLNCPGDIIPDCFFQVYQPESEELEPSNIVYLDSQINIGDIEYYEDAEGDDKIK LFGIVGSIPKTTSFTCICKKDKKSAYMTVTIDSGTKHHHHHH

[0109] Pfs230Dl / Pfs48D3 2x TG Amino Acid Sequence (SEQ ID NO:3)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNK1NGTGTGEKKVIHGCNFSSNVSSKHTFTDSED1SEVDDSAHISCNVHLSEPKYNHLVGLNCPGDIIPDCFFQVYQPESEELEPSNIVYLDSQINIGDIEYYEDAEGD DKIKLFGIVGSIPKTTSFTCICKKDKKSAYMTVTIDSGTKHHHHHH

[0110] Pfs230Dl / Pfs48D3 3x TG Amino Acid Sequence (SEQ ID NO:4)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGEKKVIHGCNFSSNVSSKHTFTDSLDISLVDDSAHISC NVHL SEPK YNHL VGLNCPGDIIPDCFF Q VYQPE SEELEP SNI V YLD S QINIGDIE Y YED AE GDDKIKLFGIVGSIPKTTSFTCICKKDKKSAYMTVTIDSGTKHHHHHH

[0111] Pfs230Dl / Pfs48D3 4x TG Amino Acid Sequence (SEQ ID NO:5)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGN RGIVEVYVEPYGNKINGTGTGTGTGEKKVIHGCNFSSNVSSKHTFTDSLDISLVDDSAHIS CNVHLSEPKYNHLVGLNCPGDIIPDCFFQVYQPESEELEPSNIVYLDSQINIGDIEYYEDAEGDDKIKLFGIVGSIPKTTSFTCICKKDKKSAYMTVTIDSGTKHHHHHH

[0112] Pfs230Dl / D9 lx TG Amino Acid Sequence (SEQ ID NO:6)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGNVHLCNFFDNPELTFDNNKIVLCK1DAELFSEVIIQLP1FGTK NVEEGVQNEEYKKFSLKPSLVFDDNNNDIKVIGKEKNEVSISLALKGVYGNRIFTFDKNG KKGEGISFFIPPIKQDTDLKFIINETIDNSNIKQRGLIYIFVRKNGTKHHHHHH

[0113] Pfs230Dl / D9 2x TG Amino Acid Sequence (SEQ ID NO:7)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGI VF. V Y VFP YGNK I NGTGTGN VHI .C NFFDNPF.I YFDNNKTVT ,CKTD AFJ T SEVTIQLPTFG TKNVEEGVQNEEYKKFSLKPSLVFDDNNNDIKVIGKEKNEVSISLALKGVYGNRIFTFDK NGKKGEGISFFIPPIKQDTDLKFIINETIDNSNIKQRGLIYIFVRKNGTKHHHHHH

[0114] Pfs230Dl / D9 3x TG Amino Acid Sequence (SEQ ID NO:8)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGNVHLCNFFDNPELTFDNNKIVLCKIDAELFSEVIIQLPIFGTKNVEEGVQNEEYKKFSLKPSLVFDDNNNDIKVIGKEKNEVSISLALKGVYGNRIFTFDKNGKKGEGISFFIPPIKQDTDLKFIINETIDNSNIKQRGLIYIFVRKNGTKHHHHHH

[0115] Pfs230Dl / D9 4x TG Amino Acid Sequence (SEQ ID NO:9)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGTGNVHECNFFDNPEETFDNNKIVECKIDAEEFSEVnOLPIFGTKNVEEGVQNEEYKKFSLKPSLVFDDNNNDIKVIGKEKNEVSISLALKGVYGNRIFTFDKNGKKGEGISFFIPPIKQDTDLKFIINETIDNSNIKQRGLIYIFVRKNGTKHHHHHH

[0116] Pfs230Dl / D7 lx TG Amino Acid Sequence (SEQ ID NO: 10)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGNRHVCDFSKNNLIVPESLKKKEELGGNPVNIHCYALLKPLDTLYVKCPTSKDNYEAAKVNISENDNEYELQVISLIEKRFHNFETLESKKPGNGDVVVHNGVVDTGPVLDNSTFEKYFKNIKIKPDKFFEKVINEYDDTEEEKDLESILPGAIVSPMKVLKKKDPFTSYAAFVVPPIVPKDLHFKVECNNTEYKDENQYISGYNGIIHIDISNSGTKHHHHHH

[0117] Pfs230Dl / D7 2x TG Amino Acid Sequence (SEQ ID NO: 11)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGNRHVCDFSKNNLIVPESLKKKEELGGNPVNIHCYALLKPLDTLYVKCPTSKDNYEAAKVNISENDNEYELQVISLIEKRFHNFETLESKKPGNGDVVVHNGVVDTGPVLDNSTFEKYFKNIKIKPDKFFEKVINEYDDTEEEKDLESILPGAIVSPMKVLKKKDPFTSYAAFVVPPIVPKDLHFKVECNNTEYKDENQYISGYNGIIHIDISNSGTKHHH HHH

[0118] Pfs230Dl / D7 3x TG Amino Acid Sequence (SEQ ID NO:12)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGNRHVCDFSKNNLIVPESLKKKEELGGNPVNIHCYALLKPLDTLYVKCPTSKDNYEAAKVNISENDNEYELQVISLIEKRFHNFETLESKKPGNGDVVVHNGVVDTGPVLDNSTFEKYFKNIKIKPDKFFEKVINEYDDTEEEKDLESILPGAIVSPMKVLKKKDPFTSYAAFVVPPIVPKDLHFKVECNNTEYKDENQYISGYNGIIHIDISNSGTKH HHHHH

[0119] Pfs230Dl / D7 4x TG Amino Acid Sequence (SEQ ID NO:13)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGTGNRHVCDFSKNNLIVPESLKKKEELGGNPVNIHCYALLKPLDTLYVKCPTSKDNYEAAKVNISENDNEYELQVISLIEKRFHNFETLESKKPGNGDVVVHNGVVDTGPVLDNSTFEKYFKNIKIKPDKFFEKVINEYDDTEEEKDLESILPGAIVSPMKVLKKKDPFTSYAAFVVPPIVPKDLHFKVECNNTEYKDENQYISGYNGIIHIDISNSGTKHHHHHH

[0120] Pfs230Dl-D2 Amino Acid Sequence (SEQ ID NO: 14)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGCAFLDEDEEEEKYGNQIEEDEHNEKIKMKTFFTQNIYKKNNIYPCYMKLYSGDIGGILFPKNIKSTTCFEEMIPYNKEIKWNKENKSLGNLVNNSVVYNKEMNAKYFNVQYVHIPTSYKDTLNLFCSIILKEEESNLISTSYLVYVSINEELNFSLFDFYESFVPIKKTIQVAGTKHHHHHH

[0121] Pfs230Dl-D3 Amino Acid Sequence (SEQ ID NO: 15)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGCAFLDEDEEEEKYGNQIEEDEHNEKIKMKTFFTQNIYKKNNIYPCYMKLYSGDIGGILFPKNIKSTTCFEEMIPYNKEIKWNKENKSLGNLVNNSVVYNKEMNAKYFNVQYVHIPTSYKDTLNLFCSnLKEEESNLISTSYLVYVSTNEELNFSLFDFYESFVPIKKTIQVAQKNVNNKEHDYTCDFTDKLDKTVPSTANGKKLFICRKHLKEFDTFTLKCNVNKTQYPNIEIFPKTLKDKKEVLKLDLDIQYQMFSKFFKFNTQNAKYLNLYPYYLIFPFNHIGKKELKNNPTYKNHKDVKYFEQSSVLSPLSSADSLGKLLNFLDTQETVCLTEKIRYLNLSINELGSDNNTFSVTFQVPPYIDIKEPFYFMFGCNNNKGEGNIGIVELLISKHHHHHHHH

[0122] Pfs230Dl / Pvs48D3 IxTG Amino Acid Sequence (SEQ ID NO: 16)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGKLIHGCNFSSDKSTHNFTNHVDMAELGENAQITCSIELVDTSYNHLIGMSCPGEVLPECFFQVYQRESPELEPSKIVYLDAQLNIGNVEYFEDSKGENIVKIFGLVGSIPKTTSFTCICRKGKKIGYMSVKGTKHHHHHH

[0123] Pvs230Dl / Pvs48D3 IxTG Amino Acid Sequence (SEQ ID NO: 17)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGQSVKGTGKLIHGCNFSSDKSTHNFTNHVDMAELGENAQI TCSIELVDTSYNHLIGMSCPGEVLPECFFQVYQRESPELEPSKIVYLDAQLNIGNVEYFED SKGENIVKIFGLVGSIPKTTSFTCICRKGKKIGYMSVKGTKHHHHHH

[0124] Pfs230Dl-4 Amino Acid Sequence (SEQ ID NO: 18)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGCAFLDEDEEEEKYGNQIEEDEHNEKIKMKTFFTQNIYKKNNIYPCYMKLYSGDIGGILFPKNIKSTTCFEEMIPYNKEIKWNKENKSLGNLVNNSVVYNKEMNAKYFNVQYVHIPTSYKDTLNLFCSIILKEEESNLISTSYLVYVSINEELNFSLFDFYESFVPIKKTIQVAQKNVNNKEHDYTCDFTDKLDKTVPSTANGKKLFICRKHLKEFDTFTLKCNVNKTQYPNIEIFPKTLKDKKEVLKLDLDIQYQMFSKFFKFNTQNAKYLNLYPYYLIFPFNHIGKKELKNNPTYKNHKDVKYFEQSSVLSPLSSADSLGKLLNFLDTQETVCLTEKIRYLNLSINELGSDNNTFSVTFQVPPYIDIKEPFYFMFGCNNNKGEGNIGIVELLISKQEEKIKGCNFHESKLDYFNENISSDTHECTLHAYENDIIGFNCLETTHPNEVEVEVEDAEIYLQPENCFNNVYKGLNSVDITTILKNAQTYNINNKKTPTFLKIPPYNLLEDVEISCQCTIKQVVKKIKVII TKNDTVLLKREVQSESTLDDKHHHHHHHH

[0125] Pfs230Dl / Pfs48D3 IxTG Nucleic Acid Sequence (SEQ ID NO: 19) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtGAGAAGAAAGTGATCCACGGCTGCAACTTCTCTTCCAATGTGAGCTCTAAGCATACCTTTACAGACAGCCTGGATATCTCTCTGGTGGACGATTCCGCTCACATCAGCTGTAACGTGCATCTGAGCGAGCCTAAGTATAACCATCTGGTGGGCCTGAATTGCCCAGGCGACATCATCCCCGATTGTTTCTTTCAGGTGTACCAGCCTGAGTCTGAGGAGCTGGAGCCATCCAACATCGTGTATCTGGATTCTCAGATCAATATCGGCGACATCGAGTACTATGAGGATGCTGAGGGCGACGATAAGATCAAGCTGTTCGGCATCGTGGGCAGCATCCCAAAGACCACATCTTTCACCTGTATCTGTAAAAAGGACAAGAAATCAGCCTATATGACTGTGACCATTGACTCAggtaccaagcaccaccatca ccatcactaa

[0126] Pfs230Dl / Pfs48D32xTG Nucleic Acid Sequence (SEQ ID NO:20) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtaccggtGAGAAGAAAGTGATCCACGGCTGCAACTTCTCTTCCAATGTGAGCTCTAAGCATACCTTTACAGACAGCCTGGATATCTCTCTGGTGGACGATTCCGCTCACATCAGCTGTAACGTGCATCTGAGCGAGCCTAAGTATAACCATCTGGTGGGCCTGAATTGCCCAGGCGACATCATCCCCGATTGTTTCTTTCAGGTGTACCAGCCTGAGTCTGAGGAGCTGGAGCCATCCAACATCGTGTATCTGGATTCTCAGATCAATATCGGCGACATCGAGTACTATGAGGATGCTGAGGGCGACGATAAGATCAAGCTGTTCGGCATCGTGGGCAGCATCCCAAAGACCACATCTTTCACCTG TATCTGTAAAAAGGACAAGAAATCAGCCTATATGACTGTGACCATTGACTCAggtacca agcaccaccatcaccatcactaa

[0127] Pfs230Dl / Pfs48D33xTG Nucleic Acid Sequence (SEQ ID NO:21) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCacagggacgggcaccggtGAGAAGAAAGTGATCCACGGCTGCAACTTCTCTTCCAATGTGAGCTCTAAGCATACCTTTACAGACAGCCTGGATATCTCTCTGGTGGACGATTCCGCTCACATCAGCTGTAACGTGCATCTGAGCGAGCCTAAGTATAACCATCTGGTGGGCCTGAATTGCCCAGGCGACATCATCCCCGATTGTTTCTTTCAGGTGTACCAGCCTGAGTCTGAGGAGCTGGAGCCATCCAACATCGTGTATCTGGATTCTCAGATCAATATCGGCGACATCGAGTACTATGAGGATGCTGAGGGCGACGATAAGATCAAGCTGTTCGGCATCGTGGGCAGCATCCCAAAGACCACATCTTTCACCTGTATCTGTAAAAAGGACAAGAAATCAGCCTATATGACTGTGACCATTGACTCAggt accaagcaccaccatcaccatcactaa

[0128] Pfs230Dl / Pfs48D3 4xTG Nucleic Acid Sequence (SEQ ID NO:22) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggaacagggacgggcaccggtGAGAAGAAAGTGATCCACGGCTGCAACTTCTCTTCCAATGTGAGCTCTAAGCATACCTTTACAGACAGCCTGGATATCTCTCTGGTGGACGATTCCGCTCACATCAGCTGTAACGTGCATCTGAGCGAGCCTAAGTATAACCATCTGGTGGGCCTGAATTGCCCAGGCGACATCATCCCCGATTGTTTCTTTCAGGTGTACCAGCCTGAGTCTGAGGAGCTGGAGCCATCCAACATCGTGTATCTGGATTCTCAGATCAATATCGGCGACATCGAGTACTATGAGGATGCTGAGGGCGACGATAAGATCAAGCTGTTCGGCATCGTGGGCAGCATCCCAAAGACCACATCTTTCACCTGTATCTGTAAAAAGGACAAGAAATCAGCCTATATGACTGTGACCATTGA CTCAggtaccaagcaccaccatcaccatcactaa

[0129] Pfs230Dl / PfsD7 IxTG Nucleic Acid Sequence (SEQ ID NO:23) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtAACAGGCACGTGTGCGACTTCTCTAAGAATAACCTGATCGTGCCTGAGAGCCTGAAGAAGAAGGAAGAGCTGGGCGGCAATCCAGTGAACATCCACTGCTATGCCCTGCTGAAGCCACTGGACACCCTGTACGTGAAGTGTCCCACATCTAAGGATAACTATGAGGCCGCCAAAGTGAATATCAGCGAGAACGACAATGAGTACGAGCTGCAGGTCATCTCCCTGATCGAGAAGCGGTTCCACAACTTCGAGACCCTGGAGAGCAAGAAGCCCGGCAACGGCGATGTGGTGGTGCACAATGGCGTGGTGGACACCGGCCCTGTGCTGGATAACTCCACATTTGAGAAGTACTTCAAGAACATCAAGATCAAGCCTGACAAGTTCTTTGAGAAAGTGATCAACGAGTATGATGACACCGAGGAGGAGAAGGATCTGGAGTCCATCCTGCCCGGCGCCATCGTGTCTCCTATGAAGGTGCTGAAGAAGAAGGACCCCTTCACCAGCTACGCAGCCTTCGTGGTGCCCCCTATCGTGCCAAAGGACCTGCACTTTAAGGTGGAGTGTAATAACACCGAGTATAAGGATGAGAACCAGTATATCTCTGGCTACAATGGCATCATCCACATCGACATCAGCAACTC Cggtaccaagcaccaccatcaccatcactaa

[0130] Pfs230Dl / PfsD7 2xTG Nucleic Acid Sequence (SEQ ID NO:24) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtaccggtAACAGGCACGTGTGCGACTTCTCTAAGAATAACCTGATCGTGCCTGAGAGCCTGAAGAAGAAGGAAGAGCTGGGCGGCAATCCAGTGAACATCCACTGCTATGCCCTGCTGAAGCCACTGGACACCCTGTACGTGAAGTGTCCCACATCTAAGGATAACTATGAGGCCGCCAAAGTGAATATCAGCGAGAACGACAATGAGTACGAGCTGCAGGTCATCTCCCTGATCGAGAAGCGGTTCCACAACTTCGAGACCCTGGAGAGCAAGAAGCCCGGCAACGGCGATGTGGTGGTGCACAATGGCGTGGTGGACACCGGCCCTGTGCTGGATAACTCCACATTTGAGAAGTACTTCAAGAACATCAAGATCAAGCCTGACAAGTTCTTTGAGAAAGTGATCAACGAGTATGATGACACCGAGGAGGAGAAGGATCTGGAGTCCATCCTGCCCGGCGCCATCGTGTCTCCTATGAAGGTGCTGAAGAAGAAGGACCCCTTCACCAGCTACGCAGCCTTCGTGGTGCCCCCTATCGTGCCAAAGGACCTGCACTTTAAGGTGGAGTGTAATAACACCGAGTATAAGGATGAGAACCAGTATATCTCTGGCTACAATGGCATCATCCACATCGACATCAGCAACTCCggtaccaagcaccaccatcaccatcactaa

[0131] Pfs230Dl / PfsD7 3xTG Nucleic Acid Sequence (SEQ ID NO:25) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCacagggacgggcaccggtAACAGGCACGTGTGCGACTTCTCTAAGAATAACCTGATCGTGCCTGAGAGCCTGAAGAAGAAGGAAGAGCTGGGCGGCAATCCAGTGAACATCCACTGCTATGCCCTGCTGAAGCCACTGGACACCCTGTACGTGAAGTGTCCCACATCTAAGGATAACTATGAGGCCGCCAAAGTGAATATCAGCGAGAACGACAATGAGTACGAGCTGCAGGTCATCTCCCTGATCGAGAAGCGGTTCCACAACTTCGAGACCCTGGAGAGCAAGAAGCCCGGCAACGGCGATGTGGTGGTGCACAATGGCGTGGTGGACACCGGCCCTGTGCTGGATAACTCCACATTTGAGAAGTACTTCAAGAACATCAAGATCAAGCCTGACAAGTTCTTTGAGAAAGTGATCAACGAGTATGATGACACCGAGGAGGAGAAGGATCTGGAGTCCATCCTGCCCGGCGCCATCGTGTCTCCTATGAAGGTGCTGAAGAAGAAGGACCCCTTCACCAGCTACGCAGCCTTCGTGGTGCCCCCTATCGTGCCAAAGGACCTGCACTTTAAGGTGGAGTGTAATAACACCGAGTATAAGGATGAGAACCAGTATATCTCTGGCTACAATGGCATCATCCACATCGACATCAGCAACTCCggtaccaagcaccaccatcaccatcactaa

[0132] Pfs230Dl / PfsD7 4xTG Nucleic Acid Sequence (SEQ ID NO:26) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggaacagggacgggcaccggtAACAGGCACGTGTGCGACTTCTCTAAGAATAACCTGATCGTGCCTGAGAGCCTGAAGAAGAAGGAAGAGCTGGGCGGCAATCCAGTGAACATCCACTGCTATGCCCTGCTGAAGCCACTGGACACCCTGTACGTGAAGTGTCCCACATCTAAGGATAACTATGAGGCCGCCAAAGTGAATATCAGCGAGAACGACAATGAGTACGAGCTGCAGGTCATCTCCCTGATCGAGAAGCGGTTCCACAACTTCGAGACCCTGGAGAGCAAGAAGCCCGGCAACGGCGATGTGGTGGTGCACAATGGCGTGGTGGACACCGGCCCTGTGCTGGATAACTCCACATTTGAGAAGTACTTCAAGAACATCAAGATCAAGCCTGACAAGTTCTTTGAGAAAGTGATCAACGAGTATGATGACACCGAGGAGGAGAAGGATCTGGAGTCCATCCTGCCCGGCGCCATCGTGTCTCCTATGAAGGTGCTGAAGAAGAAGGACCCCTTCACCAGCTACGCAGCCTTCGTGGTGCCCCCTATCGTGCCAAAGGACCTGCACTTTAAGGTGGAGTGTAATAACACCGAGTATAAGGATGAGAACCAGTATATCTCTGGCTACAATGGCATCATCCACATCGACATCAGCAACTCCggtaccaagcaccaccatcaccatcactaa

[0133] Pfs230Dl / PfsD9 IxTG Nucleic Acid Sequence (SEQ ID NO:27) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtAATGTGCACCTGTGCAATTTCTTTGACAACCCCGAGCTGACCTTCGATAACAATAAGATCGTGCTGTGCAAGATCGACGCCGAGCTGTTTAGCGAAGTGATCATCCAGCTGCCTATCTTCGGCACCAAGAACGTGGAGGAGGGCGTGCAGAATGAGGAGTACAAGAAGTTTTCTCTGAAGCCAAGCCTGGTGTTCGATGACAACAATAACGACATCAAAGTGATCGGCAAGGAGAAGAACGAGGTGTCCATCTCTCTGGCCCTGAAGGGCGTGTATGGCAACAGGATCTTTACCTTCGATAAGAATGGCAAGAAGGGCGAGGGCATCTCCTTCTTTATCCCACCCATCAAGCAGGACACAGATCTGAAGTTTATCATCAACGAGACCATCGACAACTCTAATATCAAGCAGAGGGGC CTGATCTAC ATCTTCGT GCGC AAGAAT ggtaccaagcaccaccatcaccatcactaa

[0134] Pfs230Dl / PfsD9 2xTG Nucleic Acid Sequence (SEQ ID NO:28) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtaccggtAATGTGCACCTGTGCAATTTCTTTGACAACCCCGAGCTGACCTTCGATAACAATAAGATCGTGCTGTGCAAGATCGACGCCGAGCTGTTTAGCGAAGTGATCATCCAGCTGCCTATCTTCGGCACCAAGAACGTGGAGGAGGGCGTGCAGAATGAGGAGTACAAGAAGTTTTCTCTGAAGCCAAGCCTGGTGTTCGATGACAACAATAACGACATCAAAGTGATCGGCAAGGAGAAGAACGAGGTGTCCATCTCTCTGGCCCTGAAGGGCGTGTATGGCAACAGGATCTTTACCTTCGATAAGAATGGCAAGAAGGGCGAGGGCATCTCCTTCTTTATCCCACCCATCAAGCAGGACACAGATCTGAAGTTTATCATCAACGAGACCATCGACAACTCTAATATCAAGCAGAGGGGCCTGATCTACATCTTCGTGCGCAAGAATggtaccaagcaccaccatcaccatcactaa

[0135] Pfs230Dl / PfsD9 3xTG Nucleic Acid Sequence (SEQ ID NO:29) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCacagggacgggcaccggtAATGTGCACCTGTGCAATTTCTTTGACAACCCCGAGCTGACCTTCGATAACAATAAGATCGTGCTGTGCAAGATCGACGCCGAGCTGTTTAGCGAAGTGATCATCCAGCTGCCTATCTTCGGCACCAAGAACGTGGAGGAGGGCGTGCAGAATGAGGAGTACAAGAAGTTTTCTCTGAAGCCAAGCCTGGTGTTCGATGACAACAATAACGACATCAAAGTGATCGGCAAGGAGAAGAACGAGGTGTCCATCTCTCTGGCCCTGAAGGGCGTGTATGGCAACAGGATCTTTACCTTCGATAAGAATGGCAAGAAGGGCGAGGGCATCTCCTTCTTTATCCCACCCATCAAGCAGGACACAGATCTGAAGTTTATCATCAACGAGACCATCGACAACTCTAATATCAAGCAGAGGGGCCTGATCT AC ATCTTCGT GCGC AAGAAT ggtaccaagcaccaccatcaccatcactaa

[0136] Pfs230Dl / PfsD9 4xTG Nucleic Acid Sequence (SEQ ID NO:30) accgggTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggaacagggacgggcaccggtAATGTGCACCTGTGCAATTTCTTTGACAACCCCGAGCTGACCTTCGATAACAATAAGATCGTGCTGTGCAAGATCGACGCCGAGCTGTTTAGCGAAGTGATCATCCAGCTGCCTATCTTCGGCACCAAGAACGTGGAGGAGGGCGTGCAGAATGAGGAGTACAAGAAGTTTTCTCTGAAGCCAAGCCTGGTGTTCGATGACAACAATAACGACATCAAAGTGATCGGCAAGGAGAAGAACGAGGTGTCCATCTCTCTGGCCCTGAAGGGCGTGTATGGCAACAGGATCTTTACCTTCGATAAGAATGGCAAGAAGGGCGAGGGCATCTCCTTCTTTATCCCACCCATCAAGCAGGACACAGATCTGAAGTTTATCATCAACGAGACCATCGACAACTCTAATATCAAGCAGAGGGGCCTGATCTACATCTTCGTGCGCAAGAATggtaccaagcaccaccatcaccatcactaa

[0137] Pfs230Dl / Pvs48D31xTG Nucleic Acid Sequence (SEQ ID NO:31) accggtTCTGTGCTGCAGTCTGGAGCCCTGCCTAGCGTTGGAGTCGACGAGCTGGATAAGATCGACCTGTCTTATGAGACAACCGAGTCCGGCGACACAGCTGTGTCCGAGGATAGCTATGACAAGTACGCCAGCAACAACACCAACAAGGAATACGTGTGCGACTTCACCGATCAGCTGAAGCCTACCGAGTCCGGCCCTAAAGTGAAGAAGTGTGAAGTGAAGGTGAACGAGCCTCTGATCAAAGTGAAGATTATCTGTCCACTGAAGGGCTCCGTTGAAAAGCTGTATGATAATATCGAGTACGTGCCTAAGAAGAGCCCCTACGTGGTGCTGACCAAGGAGGAAACCAAGCTGAAGGAGAAACTGCTATCCAAGCTTATCTACGGCCTGCTGATCAGCCCTACAGTGAATGAGAAAGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACCCTCCCTCCAGTGGTCCACAAGGCCACCGTGTTCTACTTTATCTGCGATAATAGCAAGACCGAGGACGACAACAAAAAGGGCAACCGGGGCATCGTGGAAGTGTACGTGGAACCTTACGGCAATAAGATCAACGGAaccggcAAGCTGATCCACGGCTGTAACTTCAGCTCTGACAAGAGCACCCACAACTTTACAAACCATGTGGACATGGCCGAGCTGGGCGAAAATGCCCAAATCACATGCAGCATCGAGCTGGTGGATACCTCCTACAACCACCTCATCGGCATGTCTTGTCCTGGAGAGGTGCTGCCTGAGTGCTTCTTTCAGGTGTATCAGCGGGAAAGCCCAGAGCTGGAACCCAGCAAGATCGTCTACCTGGACGCTCAGCTGAACATCGGCAACGTGGAATACTTCGAGGATAGCAAGGGAGAGAATATCGTGAAGATCTTCGGCCTGGTGGGCAGCATCCCTAAGACAACCTCCTTCACCTGCATCTGCAGAAAGGGCAAGAAAATTGGCTACATGAGCGTGAAAggtaccaagcaccaccatcaccatcactaa

[0138] Pvs230Dl / Pvs48D31xTG Nucleic Acid Sequence (SEQ ID NO:32) accggtGTGCTGCCTGAGGCAGGAGCATCCGACGGCGTGTTCGACAAGGTGGATGAGGCCTTTGAGACCACAATCAAGGGCGACGGCAACGTGCTGCAGGCCTCTGATCCAGAGGTGGAGACCTTTGCCAGCTCCAACACAAATAAGGAGTACGTGTGCGACTTCGTGAAGCACATCACCATGAAGGAGGCCAGCAAGAAGGTGGTCATCTGCGAGATGAAGATCCAGGAGCCACTGGTGAAGGTGAAGATCCTGTGCCCCACCAAGTACGCCGATGTGATCAAGTATGGCTCCATGGAGTTCTTTCCAAAGAAGGCCCCCTATGTGACACTGGTGAACGCCAATGAGGGCAGCGAGGAGAAGAAGGAGTCCCTGAAGGAGAAGAGGCTGGCCGCCCTGATCCACGGCGTGATCATCACACCCGAGGTGAACGAGAAGGAGAACAATTTCGAGAAGGGCGTGATCGAGTTTGTGCTGCCCCCTGTGCTGAAGGAGAAGAAGAAGCTGTACTATATCTGTGACAACGGCAAGTCCGCCGATGGCGTGTCTAATCACGGCGACAGGGGAGTGGCAGCAATCACCATCGAGCCTTACGGCCAGTCTGTGAAGGGCACAGGCac cggcAAGCTGATCCACGGCTGTAACTTCAGCTCTGACAAGAGCACCCACAACTTTACAAACCATGTGGACATGGCCGAGCTGGGCGAAAATGCCCAAATCACATGCAGCATCGAGCTGGTGGATACCTCCTACAACCACCTCATCGGCATGTCTTGTCCTGGAGAGGTGCTGCCTGAGTGCTTCTTTCAGGTGTATCAGCGGGAAAGCCCAGAGCTGGAACCCAGCAAGATCGTCTACCTGGACGCTCAGCTGAACATCGGCAACGTGGAATACTTCGAGGATAGCAAGGGAGAGAATATCGTGAAGATCTTCGGCCTGGTGGGCAGCATCCCTAAGACAACCTCCTTCACCTGCATCTGCAGAAAGGGCAAGAAAATTGGCTACATGAGCGTGAAAggtaccaagcaccaccatcaccatcactaa

[0139] Pfs230Dl-2 Nucleic Acid Sequence (SEQ ID NO:33) accggcTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCTGTGCCTTCCTGGACGAGGATGAGGAGGAGGAGAAGTACGGCAACCAGATCGAGGAGGACGAGCACAATGAGAAGATCAAGATGAAGACCTTCTTTACACAGAACATCTACAAGAAGAACAATATCTATCCCTGCTACATGAAGCTGTATTCCGGCGATATCGGCGGCATCCTGTTTCCAAAGAATATCAAGTCTACCACATGTTTCGAGGAGATGATCCCCTACAACAAGGAGATCAAGTGGAACAAGGAGAATAAGAGCCTGGGCAATCTGGTGAACAATTCCGTGGTGTACAATAAGGAGATGAACGCCAAGTACTTCAACGTGCAGTACGTGCACATCCCCACCTCTTATAAGGACACACTGAATCTGTTCTGCTCTATCATCCTGAAGGAGGAGGAGAGCAACCTGATCAGCACATCCTATCTGGTGTACGTGAGCATCAATGAGGAGCTGAACTTTAGCCTGTTCGACTTTTACGAGTCCTTCGTGCCTATCAAGAAGACCATCCAGGTGGCCggtaccaagcaccaccatcaccatc actaa

[0140] Pfs230Dl-3 Nucleic Acid Sequence (SEQ ID NO:34) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCTGTGCCTTCCTGGACGAGGATGAGGAGGAGGAGAAGTACGGCAACCAGATCGAGGAGGACGAGCACAATGAGAAGATCAAGATGAAGACCTTCTTTACACAGAACATCTACAAGAAGAACAATATCTATCCCTGCTACATGAAGCTGTATTCCGGCGATATCGGCGGCATCCTGTTTCCAAAGAATATCAAGTCTACCACATGTTTCGAGGAGATGATCCCCTACAACAAGGAGATCAAGTGGAACAAGGAGAATAAGAGCCTGGGCAATCTGGTGAACAATTCCGTGGTGTACAATAAGGAGATGAACGCCAAGTACTTCAACGTGCAGTACGTGCACATCCCCACCTCTTATAAGGACACACTGAATCTGTTCTGCTCTATCATCCTGAAGGAGGAGGAGAGCAACCTGATCAGCACATCCTATCTGGTGTACGTGAGCATCAATGAGGAGCTGAACTTTAGCCTGTTCGACTTTTACGAGTCCTTCGTGCCTATCAAGAAGACCATCCAGGTGGCCCAGAAGAACGTGAACAATAAGGAGCACGATTATACCTGTGACTTCACCGACAAGCTGGATAAGACCGTGCCATCCACAGCCAATGGCAAGAAGCTGTTCATCTGCCGGAAGCACCTGAAGGAGTTTGACACCTTCACACTGAAGTGTAACGTGAATAAGACACAGTACCCAAACATCGAGATCTTCCCCAAGACCCTGAAGGATAAGAAGGAGGTGCTGAAGCTGGACCTGGATATCCAGTATCAGATGTTTAGCAAGTTCTTTAAGTTCAACACCCAGAATGCCAAGTATCTGAATCTGTACCCTTACTATCTGATCTTCCCCTTCAACCACATCGGCAAGAAGGAGCTGAAGAACAATCCAACCTATAAGAACCACAAGGACGTGAAGTACTTTGAGCAGTCCTCTGTGCTGTCTCCACTGAGCTCCGCCGACAGCCTGGGCAAGCTGCTGAATTTCCTGGATACCCAGGAGACAGTGTGCCTGACCGAGAAGATCAGATACCTGAATCTGAGCATCAACGAGCTGGGCTCCGACAACAATACCTTTTCTGTGACATTCCAGGTGCCACCCTATATCGATATCAAGGAGCCTTTCTACTTTATGTTCGGCTGCAACAATAACAAGGGCGAGGGCAACATCGGCATCGTGGAGCTGCTGATCAGCAAGggtaccaagcaccaccatcaccatcactaa

[0141] Pfs230Dl-4 Nucleic Acid Sequence (SEQ ID NO:35) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCTGTGCCTTCCTGGACGAGGATGAGGAGGAGGAGAAGTACGGCAACCAGATCGAGGAGGACGAGCACAATGAGAAGATCAAGATGAAGACCTTCTTTACACAGAACATCTACAAGAAGAACAATATCTATCCCTGCTACATGAAGCTGTATTCCGGCGATATCGGCGGCATCCTGTTTCCAAAGAATATCAAGTCTACCACATGTTTCGAGGAGATGATCCCCTACAACAAGGAGATCAAGTGGAACAAGGAGAATAAGAGCCTGGGCAATCTGGTGAACAATTCCGTGGTGTACAATAAGGAGATGAACGCCAAGTACTTCAACGTGCAGTACGTGCACATCCCCACCTCTTATAAGGACACACTGAATCTGTTCTGCTCTATCATCCTGAAGGAGGAGGAGAGCAACCTGATCAGCACATCCTATCTGGTGTACGTGAGCATCAATGAGGAGCTGAACTTTAGCCTGTTCGACTTTTACGAGTCCTTCGTGCCTATCAAGAAGACCATCCAGGTGGCCCAGAAGAACGTGAACAATAAGGAGCACGATTATACCTGTGACTTCACCGACAAGCTGGATAAGACCGTGCCATCCACAGCCAATGGCAAGAAGCTGTTCATCTGCCGGAAGCACCTGAAGGAGTTTGACACCTTCACACTGAAGTGTAACGTGAATAAGACACAGTACCCAAACATCGAGATCTTCCCCAAGACCCTGAAGGATAAGAAGGAGGTGCTGAAGCTGGACCTGGATATCCAGTATCAGATGTTTAGCAAGTTCTTTAAGTTCAACACCCAGAATGCCAAGTATCTGAATCTGTACCCTTACTATCTGATCTTCCCCTTCAACCACATCGGCAAGAAGGAGCTGAAGAACAATCCAACCTATAAGAACCACAAGGACGTGAAGTACTTTGAGCAGTCCTCTGTGCTGTCTCCACTGAGCTCCGCCGACAGCCTGGGCAAGCTGCTGAATTTCCTGGATACCCAGGAGACAGTGTGCCTGACCGAGAAGATCAGATACCTGAATCTGAGCATCAACGAGCTGGGCTCCGACAACAATACCTTTTCTGTGACATTCCAGGTGCCACCCTATATCGATATCAAGGAGCCTTTCTACTTTATGTTCGGCTGCAACAATAACAAGGGCGAGGGCAACATCGGCATCGTGGAGCTGCTGATCAGCAAGAAGCAGGAGGAGAAGATCAAGGGCTGTAATTTTCACGAGTCCAAGCTGGACTACTTCAACGAGAATATCTCTAGCGATACCCACGAGTGCACACTGCACGCCTATGAGAATGACATCATCGGCTTTAACTGTCTGGAGACCACACACCCTAACGAAGTGGAGGTGGAGGTGGAGGATGCCGAGATCTACCTGCAGCCAGAGAATTGCTTCAATAACGTGTATAAGGGCCTGAACTCTGTGGACATCACCACAATCCTGAAGAATGCCCAGACCTACAACATCAACAACAAGAAGACCCCAACATTCCTGAA GATCCCTCCATATAACCTGCTGGAGGACGTGGAGATCAGCTGCCAGTGTACAATCAA GCAGGTGGTGAAGAAGATCAAAGTGATCATCACCAAGAATGATACAGTGCTGCTGA AGCGGGAGGTGCAGTCTGAGAGCACCCTGGATGACAAGggtaccaagcaccaccatcaccatcacta a

[0142] Pvs230Dl Amino Acid Sequence (SEQ ID NO:36)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHI TMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGS EEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSA DGVSNHGDRGVAAITIEP YGQ S VKG

[0143] Pvs230D2 Nucleic Acid Sequence (SEQ ID NO:37) accgggTCTGTGAAGGGCTGTAATTATACCGGCAAGGGCAAGCACTTCTTTAGCTACGA CTATGAGGAGGGCGCCGATATGAAGCACCCTTGCGTGTTCCAGCTGAACGCCGGCGAGATCGGCGGCATCATGTTTCCCGAGGGCACAAAGAGCACCTCCTGTTTCGACAGGATGGTGCCTCACAGCGCCAATGAGAAGTGGGATAAGAAGAAGAAGAGCCTGAGCC AGGTCATCAACGGCGCCGTGGTGTACAACAATGATTTCAAGGCCAAGTACTTTACCGTGAAGTATTTCTCTATCCCTAAGAGCTTTCGCGAGGCCACACACCTGATGTGCCACATCCAGCTGACCGACGGCGATAAGAAGCACATGGTGTACATCTCCATCAATCAGGAG ggtaccaagcaccaccatcaccatcactaa

[0144] Pvs230D2 Amino Acid Sequence (SEQ ID NO:38)TGSVKGCNYTGKGKHFFSYDYEEGADMKHPCVFQLNAGEIGGIMFPEGTKSTSCFDRMVPHSANEKWDKKKKSLSQVINGAVVYNNDFKAKYFTVKYFSIPKSFREATHLMCHIQLTDGDKKHMVYISINQEGTKHHHHHH

[0145] Pfs230Dl / Pvs230D2 lx TG Nucleic Acid Sequence (SEQ ID NO:39) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtTCTGTGAAGGGCTGTAATTATACCGGCAAGGGCAAGCACTTCTTTAGCTACGACTATGAGGAGGGCGCCGATATGAAGCACCCTTGCGTGTTCCAGCTGAACGCCGGCGAGATCGGCGGCATCATGTTTCCCGAGGGCACAAAGAGCACCTCCTGTTTCGACAGGATGGTGCCTCACAGCGCCAATGAGAAGTGGGATAAGAAGAAGAAGAGCCTGAGCCAGGTCATCAACGGCGCCGTGGTGTACAACAATGATTTCAAGGCCAAGTACTTTACCGTGAAGTATTTCTCTATCCCTAAGAGCTTTCGCGAGGCCACACACCTGATGTGCCACATCCAGCTGACCGACGGCGATAAGAAGCACAT GGTGT AC ATCTCC ATC AATC AGGAGggtaccaagcaccaccatcaccatcactaa

[0146] Pfs230Dl / Pvs230D2 lx TG Amino Acid Sequence (SEQ ID NO:40)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGSVKGCNYTGKGKHFFSYDYEEGADMKHPCVFQLNAGEIG GIMFPEGTKSTSCFDRMVPHSANEKWDKKKKSLSQVINGAVVYNNDFKAKYFTVKYFS IPKSFREATHLMCHIQLTDGDKKHMVYISINQEGTKHHHHHH

[0147] Pvs230D3 Nucleic Acid Sequence (SEQ ID NO:41) accgggAAGGAGTATACATGTGACTTCACCGATCAGCTGGACAAGAAGGCCGGAGAGTCCGAGAAGAAAGTGATCATCTGCAGGAAGCGCCTGACCGAGTTCGACTCTTTTAATATGAAGTGTGCCATCAACAAGGCCAAGTACGCCAATCTGGAGATGATCCCAAAGATGCTGAAGGAGGAGAAGAAGAAGAAGAAGAACGTGCTGAAGGTGCAGCTGGACGTGCAGTACGAGCTGTTTAAGAAGTATTTCCACTTTCACGATAAGTACGGCCGGTATCTGCCAGGCTATCCCAGCTTCTTTTCCTTCCCACTGAACAGAGTGGCCAAGCTGGAGCTGAAGAAGAATTCCCTGTTTAAGAACCACAAGGACTCTAGCTACTTCGATGAGGTGGCCTCCCCCTCTGACGGCTTTGTGAAGCTGCTGTCCTTCCTGGATGCCCAGGACACAGTGATCCTGAACGAGCAGGTGTCTGACCTGACAATCTCCACCGAGCAGTCTACAGAGGATCTGTTTACCCTGCAGCTGCAGATCCCTCCATACATCACCACAAATGAGCCTCTGTATTTCCTGATCGGCTGCAACAATACCAAGGACGGCGGCAACCTGGGCATCGTGGAGCT GGT C ATC AGC AAGAAT ggtaccaagcaccaccatcaccatcactaa

[0148] Pvs230D3 Amino Acid Sequence (SEQ ID NO:42)TGKEYTCDFTDQLDKKAGESEKKVIICRKRLTEFDSFNMKCAINKAKYANLEMIPKMLK EEKKKKKNVLKVQLD VQ YELFKKYFHFHDKYGRYLPGYP SFF SFPLNRVAKLELKKNS LFKNHKDSSYFDEVASPSDGFVKLLSFLDAQDTVILNEQVSDLTISTEQSTEDLFTLQLQI PPYITTNEPLYFLIGCNNTKDGGNLGIVELVISKNGTKHHHHHH

[0149] Pfs230Dl / Pvs230D3 IxTG Nucleic Acid Sequence (SEQ ID NO:43) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtAAGGAGTATACATGTGACTTCACCGATCAGCTGGACAAGAAGGCCGGAGAGTCCGAGAAGAAAGTGATCATCTGCAGGAAGCGCCTGACCGAGTTCGACTCTTTTAATATGAAGTGTGCCATCAACAAGGCCAAGTACGCCAATCTGGAGATGATCCCAAAGATGCTGAAGGAGGAGAAGAAGAAGAAGAAGAACGTGCTGAAGGTGCAGCTGGACGTGCAGTACGAGCTGTTTAAGAAGTATTTCCACTTTCACGATAAGTACGGCCGGTATCTGCCAGGCTATCCCAGCTTCTTTTCCTTCCCACTGAACAGAGTGGCCAAGCTGGAGCTGAAGAAGAATTCCCTGTTTAAGAACCACAAGGACTCTAGCTACTTCGATGAGGTGGCCTCCCCCTCTGACGGCTTTGTGAAGCTGCTGTCCTTCCTGGATGCCCAGGACACAGTGATCCTGAACGAGCAGGTGTCTGACCTGACAATCTCCACCGAGCAGTCTACAGAGGATCTGTTTACCCTGCAGCTGCAGATCCCTCCATACATCACCACAAATGAGCCTCTGTATTTCCTGATCGGCTGCAACAATACCAAGGACGGCGGCAACCTGGGCATCGTGGAGCTGGTCATCAGCAAGAATggtaccaagcaccaccatcacc atcactaa

[0150] Pfs230Dl / Pvs230D3 IxTG Amino Acid Sequence (SEQ ID NO:44)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKL KEKLL SKLI YGLLISPT VNEKENNFKEGVIEFTLPP VVHKAT VF YFICDN SKTEDDNKKGNRGIVEVYVEPYGNKINGTGKEYTCDFTDQLDKKAGESEKKVIICRKRLTEFDSFNMKCAINKAKYANLEMIPKMLKEEKKKKKNVLKVQLDVQYELFKKYFHFHDKYGRYLPGYPSF FSFPLNRVAKLELKKNSLFKNHKDSSYFDEVASPSDGFVKLLSFLDAQDTVILNEQVSDL TISTEQSTEDLFTLQLQIPPYITTNEPLYFLIGCNNTKDGGNLGIVELVISKNGTKHHHHHH

[0151] Pvs230D4 Nucleic Acid Sequence (SEQ ID NO:45) accgggCTGGTGAAGGGCTGTAACTTTTCCGATGGCAAGATGGAGCACTTCACAAACAATGTGGGCAGCACCGGCGAGAAGTGCAACATCACAGCCTTTCCAAATGACGTGGTGGGCTTCAACTGTAGCAAGACAGTGCTGCCACCTTCCGACGAGGCAGAGGATTCTGCCACCAGCAACGTGGTCATCGAGCCTGAGGGCTGCTTTACACAGGTGTATTCCTCTAGCCAGAAGACCGATATCGTGTCTATCCTGCCAGGCGTGCTGCTGTACACAAGCCGGGCCAAGTCCTCTAAGGCCTTCATCAAGATCCCACCCGTGCTGCTGAAGGACAAGTTCACCTTTTCTTGCAAGTGTAAGATCGAGAGCGTGCAGAATGAGATGGAGGTGAACGTGAGGCGCGAGAGAGAGTCCAAGGAGGCCGTGATCGCCGAGCTGAAGCAGGAGggtaccaa gcaccaccatcaccatcactaa

[0152] Pvs230D4 Amino Acid Sequence (SEQ ID NO:46)TGLVKGCNFSDGKMEHFTNNVGSTGEKCNITAFPNDVVGFNCSKTVLPPSDEAEDSATS NVVIEPEGCFTQ VYS S SQKTDIVSILPGVLL YTSRAKS SKAFIKIPPVLLKDKFTFSCKCKIE SVQNEMEVNVRRERESKEAVIAELKQEGTKHHHHHH

[0153] Pfs230Dl / Pvs230D4 lx TG Nucleic Acid Sequence (SEQ ID NO:47) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtCTGGTGAAGGGCTGTAACTTTTCCGA TGGCAAGATGGAGCACTTCACAAACAATGTGGGCAGCACCGGCGAGAAGTGCAACA TCACAGCCTTTCCAAATGACGTGGTGGGCTTCAACTGTAGCAAGACAGTGCTGCCACCTTCCGACGAGGCAGAGGATTCTGCCACCAGCAACGTGGTCATCGAGCCTGAGGGCTGCTTTACACAGGTGTATTCCTCTAGCCAGAAGACCGATATCGTGTCTATCCTGCCAGGCGTGCTGCTGTACACAAGCCGGGCCAAGTCCTCTAAGGCCTTCATCAAGATCCCA CCCGTGCTGCTGAAGGACAAGTTCACCTTTTCTTGCAAGTGTAAGATCGAGAGCGTG CAGAATGAGATGGAGGTGAACGTGAGGCGCGAGAGAGAGTCCAAGGAGGCCGTGATCGCCGAGCTGAAGCAGGAGggtaccaagcaccaccatcaccatcactaa

[0154] Pfs230Dl / Pvs230D4 lx TG Pvs230D4 Amino Acid Sequence (SEQ ID NO:48)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLL SKLIYGLLISPT VNEKENNFKEGVIEFTLPP VVHKAT VF YFICDN SKTEDDNKKGN RGIVEVYVEPYGNKTNGTGLVKGCNFSDGKMEHFTNNVGSTGEKCNITAFPNDVVGFN C SKT VLPP SDE AEDS AT SNVVIEPEGCFTQ VYS S SQKTDIVSILPGVLLYT SRAKS SKAFIK IPPVLLKDKFTFSCKCKIESVQNEMEVNVRRERESKEAVIAELKQEGTKHHHHHH

[0155] Pvs230D5 Nucleic Acid Sequence (SEQ ID NO:49) accgggATGATCTATAGCTGCGCCAATAAGACACTGATCGAGCCTAAGCTGGAGAAGCTGGACGATAGGTACGTGAAGAACACCTGCCACGTGGACGCCAAGGAGATGCACTCCTATGTGGAGCTGTTTTGTCCAAGCCGCGATCTGTCCATCCACCAGAACATCAATCTGTACTTCAATACAGTGAAGCCCACCAAGGTGGAGAGCCCTAAGGTGCTGAACCAGGTGGAGCTGGAGAAGCTGATCCCACACGCCGAGATGCTGCACAAGACAAGGAGCATCCTGCCCCTGCAGTCCTGCACCTCTGGCAAGGAGATGGTGGACATCTCCCACGTGTATCTGTTCTTTCCCTACTATGTGAAAGAGGAGCAGGAGTTTAACATCCTGTGCGATAACA GCAATACAGTGTTCGAGGGCAAGAGGGGCGGCCTGCTGACCTACCAGGTGAAGGTGCCTAAGCGCggtaccaagcaccaccatcaccatcactaa

[0156] Pvs230D5 Amino Acid Sequence (SEQ ID NO:50)TGMIYSCANKTLIEPKLEKLDDRYVKNTCHVDAKEMHSYVELFCPSRDLSIHQNINLYFNTVKPTKVESPKVLNQVELEKLIPHAEMLHKTRSILPLQSCTSGKEMVDISHVYLFFPYYVKEEQEFNILCDNSNTVFEGKRGGLLTYQVKVPKRGTKHHHHHH

[0157] Pfs230Dl / Pvs230D5 lx TG Nucleic Acid Sequence (SEQ ID NO:51) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtATGATCTATAGCTGCGCCAATAAGACACTGATCGAGCCTAAGCTGGAGAAGCTGGACGATAGGTACGTGAAGAACACCTGCCACGTGGACGCCAAGGAGATGCACTCCTATGTGGAGCTGTTTTGTCCAAGCCGCGATCTGTCCATCCACCAGAACATCAATCTGTACTTCAATACAGTGAAGCCCACCAAGGTGGAGAGCCCTAAGGTGCTGAACCAGGTGGAGCTGGAGAAGCTGATCCCACACGCCGAGATGCTGCACAAGACAAGGAGCATCCTGCCCCTGCAGTCCTGCACCTCTGGCAAGGAGATGGTGGACATCTCCCACGTGTATCTGTTCTTTCCCTACTATGTGAAAGAGGAGCAGGAGTTTAACATCCTGTGCGATAACAGCAATACAGTGTTCGAGGGCAAGAGGGGCGGCCTGCTGACCTACCAGGTGAAGGTGCCTAAGCGCggtaccaagcaccaccatcaccatcactaa

[0158] Pfs230Dl / Pvs230D5 lx TG Amino Acid Sequence (SEQ ID NO:52)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLL SKLIYGLLISPT VNEKENNFKEGVIEFTLPP VVHK AT VF YFICDN SKTEDDNKKGNRGIVEVYVEPYGNKINGTGMIYSCANKTLIEPKLEKLDDRYVKNTCHVDAKEMHSYVELFCPSRDLSIHQNINLYFNTVKPTKVESPKVLNQVELEKLIPHAEMLHKTRSILPLQSCTSGKEMVDISHVYLFFPYYVKEEQEFNILCDNSNTVFEGKRGGLLTYQVKVPKRGTKHHH HHH

[0159] Pvs230D2-3 Nucleic Acid Sequence (SEQ ID NO:53) accgggTCTGTGAAGGGCTGTAATTATACCGGCAAGGGCAAGCACTTCTTTAGCTACGACTATGAGGAGGGCGCCGATATGAAGCACCCTTGCGTGTTCCAGCTGAACGCCGGCGAGATCGGCGGCATCATGTTTCCCGAGGGCACAAAGAGCACCTCCTGTTTCGACAGGATGGTGCCTCACAGCGCCAATGAGAAGTGGGATAAGAAGAAGAAGAGCCTGAGCCAGGTCATCAACGGCGCCGTGGTGTACAACAATGATTTCAAGGCCAAGTACTTTACCGTGAAGTATTTCTCTATCCCTAAGAGCTTTCGCGAGGCCACACACCTGATGTGCCACATCCAGCTGACCGACGGCGATAAGAAGCACATGGTGTACATCTCCATCAATCAGGAGCTGAACCTGAGCGTGTTCGACCTGTATGAGGCCTTCGCCAACGTGAAGAAAGTGATCCAGATCGCCAAGCAGACAGACGATAAGAAGGAGTATACATGTGACTTCACCGATCAGCTGGACAAGAAGGCCGGAGAGTCCGAGAAGAAAGTGATCATCTGCAGGAAGCGCCTGACCGAGTTCGACTCTTTTAATATGAAGTGTGCCATCAACAAGGCCAAGTACGCCAATCTGGAGATGATCCCAAAGATGCTGAAGGAGGAGAAGAAGAAGAAGAAGAACGTGCTGAAGGTGCAGCTGGACGTGCAGTACGAGCTGTTTAAGAAGTATTTCCACTTTCACGATAAGTACGGCCGGTATCTGCCAGGCTATCCCAGCTTCTTTTCCTTCCCACTGAACAGAGTGGCCAAGCTGGAGCTGAAGAAGAATTCCCTGTTTAAGAACCACAAGGACTCTAGCTACTTCGATGAGGTGGCCTCCCCCTCTGACGGCTTTGTGAAGCTGCTGTCCTTCCTGGATGCCCAGGACACAGTGATCCTGAACGAGCAGGTGTCTGACCTGACAATCTCCACCGAGCAGTCTACAGAGGATCTGTTTACCCTGCAGCTGCAGATCCCTCCATACATCACCACAAATGAGCCTCTGTATTTCCTGATCGGCTGCAACAATACCAAGGACGGCGGCAACCTGGGCATCGTGGAGCTGGTCATCAGCAAGAATggtaccaagcaccaccatcaccatcactaa

[0160] Pvs230D2-3 Amino Acid Sequence (SEQ ID NO:54)TGSVKGCNYTGKGKHFFSYDYEEGADMKHPCVFQLNAGEIGGIMFPEGTKSTSCFDRMVPHSANEKWDKKKKSLSQVINGAVVYNNDFKAKYFTVKYFSIPKSFREATHLMCHIQLTDGDKKHMVYISINQELNLSVFDLYEAFANVKKVIQIAKQTDDKKEYTCDFTDQLDKKAGESEKKVIICRKRLTEFDSFNMKCAINKAKYANLEMIPKMLKEEKKKKKNVLKVQLDVQYELFKKYFHFHDKYGRYLPGYPSFFSFPLNRVAKLELKKNSLFKNHKDSSYFDEVASPSDGFVKLLSFLDAQDTVILNEQVSDLTISTEQSTEDLFTLQLQIPPYITTNEPLYFLIGCNNTKDGGNLGIVELVISKNGTKHHHHHH

[0161] Pfs230Dl / Pvs230D2-3 lx TG Nucleic Acid Sequence (SEQ ID NO:55) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtTCTGTGAAGGGCTGTAATTATACCGGCAAGGGCAAGCACTTCTTTAGCTACGACTATGAGGAGGGCGCCGATATGAAGCACCCTTGCGTGTTCCAGCTGAACGCCGGCGAGATCGGCGGCATCATGTTTCCCGAGGGCACAAAGAGCACCTCCTGTTTCGACAGGATGGTGCCTCACAGCGCCAATGAGAAGTGGGATAAGAAGAAGAAGAGCCTGAGCCAGGTCATCAACGGCGCCGTGGTGTACAACAATGATTTCAAGGCCAAGTACTTTACCGTGAAGTATTTCTCTATCCCTAAGAGCTTTCGCGAGGCCACACACCTGATGTGCCACATCCAGCTGACCGACGGCGATAAGAAGCACATGGTGTACATCTCCATCAATCAGGAGCTGAACCTGAGCGTGTTCGACCTGTATGAGGCCTTCGCCAACGTGAAGAAAGTGATCCAGATCGCCAAGCAGACAGACGATAAGAAGGAGTATACATGTGACTTCACCGATCAGCTGGACAAGAAGGCCGGAGAGTCCGAGAAGAAAGTGATCATCTGCAGGAAGCGCCTGACCGAGTTCGACTCTTTTAATATGAAGTGTGCCATCAACAAGGCCAAGTACGCCAATCTGGAGATGATCCCAAAGATGCTGAAGGAGGAGAAGAAGAAGAAGAAGAACGTGCTGAAGGTGCAGCTGGACGTGCAGTACGAGCTGTTTAAGAAGTATTTCCACTTTCACGATAAGTACGGCCGGTATCTGCCAGGCTA TCCCAGCTTCTTTTCCTTCCCACTGAACAGAGTGGCCAAGCTGGAGCTGAAGAAGAA TTCCCTGTTTAAGAACCACAAGGACTCTAGCTACTTCGATGAGGTGGCCTCCCCCTC TGACGGCTTTGTGAAGCTGCTGTCCTTCCTGGATGCCCAGGACACAGTGATCCTGAA CGAGCAGGTGTCTGACCTGACAATCTCCACCGAGCAGTCTACAGAGGATCTGTTTAC CCTGCAGCTGCAGATCCCTCCATACATCACCACAAATGAGCCTCTGTATTTCCTGATCGGCTGCAACAATACCAAGGACGGCGGCAACCTGGGCATCGTGGAGCTGGTCATCA GCAAGAATggtaccaagcaccaccatcaccatcactaa

[0162] Pfs230Dl / Pvs230D2-3 lx TG Amino Acid Sequence (SEQ ID NO:56)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKL KEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGN RGIVEVYVEPYGNKINGTGSVKGCNYTGKGKHFFSYDYEEGADMKHPCVFQLNAGEIG GIMFPEGTKSTSCFDRMVPHSANEKWDKKKKSLSQVINGAVVYNNDFKAKYFTVKYFS IPKSFREATHLMCHIQLTDGDKKHMVYISINQELNLSVFDLYEAFANVKKVIQIAKQTDDKKEYTCDFTDQLDKKAGESEKKVIICRKRLTEFDSFNMKCAINKAKYANLEMIPKMLKE EKKKKKNVLKVQLDVQYELFKKYFHFHDKYGRYLPGYPSFFSFPLNRVAKLELKKNSL FKNHKDSSYFDEVASPSDGFVKLLSFLDAQDTVILNEQVSDLTISTEQSTEDLFTLQLQIP PYITTNEPLYFLIGCNNTKDGGNLGIVELVISKNGTKHHHHHH

[0163] Pvs230D13-14 Nucleic Acid Sequence (SEQ ID NO:57) accgggAGCTGCTCCTACGACTCCGTGAAGAATGTGACAACCTGTAACGTGGTGGAGTT TATGGAGTCCCTGGCCCAGAAGGACAATAAGACAAACCACTACTCTGTGCAGCTGGCCAAGTGGGATAAGCTGATCCTGAAGTATCCAACCAATGAGAAGGAGAACTACGAGAAGGTGTATATCAATCCCATCAACATCAAGGACAAGGTGCTGTTCAGAAAGGTGCC CACCCACATCGAGGACCTGCTGCCTGGCGCCATCACAACCAATAAGCTGGATAGCAGGACAAAGATCATCGAGTACACCCTGCGCGTGCCTCCATACGTGAGCAAGGAGGTGCACTTCTCTATCGAGTTTAACAATTCCCTGACCTCTAAGATGGTGAACTACAATGTG GCCTATGGCGGCGTGGTGGAGATCTTTATCCACGTGAGGGAGGGCTACACAGAGATCTCTGGCTGCGACTTCACCGGCCAGTATAACCACCTGTTCAGCCACAATTTTGCCCC CAACCTGAAGGAGAGCAAGACATGTTCCGTGGTGTTTGGCAACAATTCCTATGCCGG CTTTGCCTGCCCTACCAAGTTCGAGATCGTGCCACACAATTGTTTCGCCAGCGTGTA CGACCGCAACGATGGCGAGAAGGTGAAGAAGCTGGTGGACCTGAGCGAGGAGGCC GAGTACGATTTTGTGCAGTATAATTCCCGGGGCGTGTCTCTGAGCTATCTGACCTTC AAGAGAGATACAAAGACCCACTCCGTGTCTTGCAAGTGCGTGAGCCCTCAGGTGAC ACACACCATCAACGTGACATTCGAGCCCGACGCCGATCACTCTCTGCCTAAGACCCG GATCAGAATCAGGTACTTTGACCTGAGCCGGGCCTCCTTCAGCTCCCACCTGAGAGGCGGCggtaccaagcaccaccatcaccatcactaa

[0164] Pvs230D13-14 Amino Acid Sequence (SEQ ID NO:58)TGSCSYDSVKNVTTCNVVEFMESLAQKDNKTNHYSVQLAKWDKLILKYPTNEKENYE K V YINPINIKDK VLFRK VPTHIEDLLPGAITTNKLD SRTKIIEY TLRVPP Y V SKE VHF SIEFN NSLTSKMVNYNVAYGGVVEIFIHVREGYTEISGCDFTGQYNHLFSHNFAPNLKESKTCS VVFGNNSYAGFACPTKFEIVPHNCFASVYDRNDGEKVKKLVDLSEEAEYDFVQYNSRG VSLSYLTFKRDTKTHSVSCKCVSPQVTHTINVTFEPDADHSLPKTRIRIRYFDLSRASFSS HLRGGGTKHHHHHH

[0165] Pfs230Dl / Pvs230D13-14 lx TG Nucleic Acid Sequence (SEQ ID NO:59) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAA GATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATA GCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACC GACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGT GAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGA AGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCA AGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTG ATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTT CACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAG CAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTG GAGCCATACGGCAACAAGATCAATGGCaccggtAGCTGCTCCTACGACTCCGTGAAGAATGTGACAACCTGTAACGTGGTGGAGTTTATGGAGTCCCTGGCCCAGAAGGACAATAAGACAAACCACTACTCTGTGCAGCTGGCCAAGTGGGATAAGCTGATCCTGAAGTATCCAACCAATGAGAAGGAGAACTACGAGAAGGTGTATATCAATCCCATCAACATCAAGGACAAGGTGCTGTTCAGAAAGGTGCCCACCCACATCGAGGACCTGCTGCCTGGCGCCATCACAACCAATAAGCTGGATAGCAGGACAAAGATCATCGAGTACACCCTGCGCGTGCCTCCATACGTGAGCAAGGAGGTGCACTTCTCTATCGAGTTTAACAATTCCCTGACCTCTAAGATGGTGAACTACAATGTGGCCTATGGCGGCGTGGTGGAGATCTTTATCCACGTGAGGGAGGGCTACACAGAGATCTCTGGCTGCGACTTCACCGGCCAGTATAACCACCTGTTCAGCCACAATTTTGCCCCCAACCTGAAGGAGAGCAAGACATGTTCCGTGGTGTTTGGCAACAATTCCTATGCCGGCTTTGCCTGCCCTACCAAGTTCGAGATCGTGCCACACAATTGTTTCGCCAGCGTGTACGACCGCAACGATGGCGAGAAGGTGAAGAAGCTGGTGGACCTGAGCGAGGAGGCCGAGTACGATTTTGTGCAGTATAATTCCCGGGGCGTGTCTCTGAGCTATCTGACCTTCAAGAGAGATACAAAGACCCACTCCGTGTCTTGCAAGTGCGTGAGCCCTCAGGTGACACACACCATCAACGTGACATTCGAGCCCGACGCCGATCACTCTCTGCCTAAGACCCGGATCAGAATCAGGTACTTTGACCTGAGCCGGGCCTCCTTCAGCTCCCACCTGAGAGGCGGCggtaccaagcaccaccatcaccatcactaa[001661 Pfs230Dl / Pvs230D13-14 lx TG Amino Acid Sequence (SEQ ID NO:60)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLL SKLI YGLLISPT VNEKENNFKEGVIEFTLPP VVHK AT VF YFICDN SKTEDDNKKGNRGIVEVYVEPYGNKINGTGSCSYDSVKNVTTCNVVEFMESLAQKDNKTNHYSVQLAKWDKLILKYPTNEKENYEKVYINPINIKDKVLFRKVPTHIEDLLPGAITTNKLDSRTKIIEYTLRVPPYVSKEVHFSIEFNNSLTSKMVNYNVAYGGVVEIFIHVREGYTEISGCDFTGQYNHLFSHNFAPNLKESKTCSVVFGNNSYAGFACPTKFEIVPHNCFASVYDRNDGEKVKKLVDLSEEAEYDFVQYNSRGVSLSYLTFKRDTKTHSVSCKCVSPQVTHTINVTFEPDADHSLPKTRIRIRYFDLSRASFSSFrLRGGGTKHHHHHH

[0167] Pvs230D7 Nucleic Acid Sequence (SEQ ID NO:61) accgggCTGTGCGATTTCTCCAAGAAGTCTCTGATCGTGCCCGAGCCTCTGACAGAGGAGTCTCCACCCGACATCCACTGCTACAGCGCCCTGAAGCCTCTGGATACACTGTATGTGAAGTGTCCAACCGAGAAGGCCGCCTACGAGGTGGCCAAGGGCAAGACCGGCGAGGAGGACGAGGAGGAGGCAAAGGGCATCATCGCCCTGCTGGAGGGCGATCCAGCAGACGCAGGCGATCCATCCGCCCCTCCACTGGACGATGCCTCTTTCGTGAAGTATTTTGAGAAGGTGTCCATGAAGCCCTCTGGCTTCTTTAAGAACGTGCTGAATGAGGACGGCGATAAGGAGGAGGAGATCGAGAGGGTGCTGCCTGGAGCAGCCAATACATCTATGATCGTGCTGAAGAAGAAGGATCCATTCACCAGCTACGCCGCCGTGATCATCCCCCCTTCTGTGAGCAGGAATACCTTCTTTAAGGTGCAGTGCAACAATGACGAGTACAAGGAGGGCGCCAAGAACGGCGGCTATAAGGGCGTGATCCACCTGGACATCTCCCGCTCTGAGg gtaccaagcaccaccatcaccatcactaa

[0168] Pvs230D7 Amino Acid Sequence (SEQ ID NO:62)TGLCDFSKKSLIVPEPLTEESPPDIHCYSALKPLDTLYVKCPTEKAAYEVAKGKTGEEDEEEAKGIIALLEGDPADAGDPSAPPLDDASFVKYFEKVSMKPSGFFKNVLNEDGDKEEEIERVLPGAANTSMIVLKKKDPFTSYAAVIIPPSVSRNTFFKVQCNNDEYKEGAKNGGYKGV IHLDISRSEGTKHHHHHH

[0169] Pfs230Dl / Pvs230D7 IxTG Nucleic Acid Sequence (SEQ ID NO:63) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtCTGTGCGATTTCTCCAAGAAGTCTCTGATCGTGCCCGAGCCTCTGACAGAGGAGTCTCCACCCGACATCCACTGCTACAGCGCCCTGAAGCCTCTGGATACACTGTATGTGAAGTGTCCAACCGAGAAGGCCGCCTACGAGGTGGCCAAGGGCAAGACCGGCGAGGAGGACGAGGAGGAGGCAAAGGGCATCATCGCCCTGCTGGAGGGCGATCCAGCAGACGCAGGCGATCCATCCGCCCCTCCACTGGACGATGCCTCTTTCGTGAAGTATTTTGAGAAGGTGTCCATGAAGCCCTCTGGCTTCTTTAAGAACGTGCTGAATGAGGACGGCGATAAGGAGGAGGAGATCGAGAGGGTGCTGCCTGGAGCAGCCAATACATCTATGATCGTGCTGAAGAAGAAGGATCCATTCACCAGCTACGCCGCCGTGATCATCCCCCCTTCTGTGAGCAGGAATACCTTCTTTAAGGTGCAGTGCAACAATGACGAGTACAAGGAGGGCGCCAAGAACGGCGGCTATAAGGGCGTGATCCACCTGGACATCTCCCGCTCTGAGggtaccaagcaccaccatcaccatcactaa

[0170] Pfs230Dl / Pvs230D7 IxTG Amino Acid Sequence (SEQ ID NO:64)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGLCDFSKKSLIVPEPLTEESPPDIHCYSALKPLDTLYVKCPTEKAAYEVAKGKTGEEDEEEAKGIIALLEGDPADAGDPSAPPLDDASFVKYFEKVSMKPSGFFKNVLNEDGDKEEEIERVLPGAANTSMIVLKKKDPFTSYAAVIIPPSVSRNTFFKVQCNNDEYKEGAKNGGYKGVIHLDISRSEGTKHHHHHH

[0171] Pvs230Dl / Pvs230D7 IxTG Nucleic Acid Sequence (SEQ ID NO:65) accgggGTGCTGCCAGAGGCAGGAGCATCCGATGGCGTGTTTGACAAGGTGGATGAGGCCTTCGAGACCACAATCAAGGGCGACGGAAACGTGCTGCAGGCAAGCGATCCAGAGGTGGAGACATTTGCCAGCTCCAACACCAATAAGGAGTACGTGTGCGATTTCGTGAAGCACATCACCATGAAGGAGGCCTCCAAGAAGGTGGTCATCTGCGAGATGAAGATCCAGGAGCCACTGGTGAAGGTGAAGATCCTGTGCCCCACAAAGTACGCCGACGTGATCAAGTATGGCAGCATGGAGTTCTTTCCTAAGAAGGCCCCATACGTGACCCTGGTGAACGCCAATGAGGGCTCTGAGGAGAAGAAGGAGAGCCTGAAGGAGAAGCGGCTGGCCGCCCTGATCCACGGCGTGATCATCACCCCCGAGGTGAACGAGAAGGAGAACAATTTTGAGAAGGGCGTGATCGAGTTCGTGCTGCCACCCGTGCTGAAGGAGAAGAAGAAGCTGTACTATATCTGCGACAATGGCAAGTCTGCCGATGGCGTGAGCAACCACGGCGACAGAGGAGTGGCAGCAATCACAATCGAGCCCTACGGCCAGTCTGTGAAGGGCaccggaCTGTGCGATTTCTCCAAGAAGTCTCTGATCGTGCCCGAGCCTCTGACAGAGGAGTCTCCACCCGACATCCACTGCTACAGCGCCCTGAAGCCTCTGGATACACTGTATGTGAAGTGTCCAACCGAGAAGGCCGCCTACGAGGTGGCCAAGGGCAAGACCGGCGAGGAGGACGAGGAGGAGGCAAAGGGCATCATCGCCCTGCTGGAGGGCGATCCAGCAGACGCAGGCGATCCATCCGCCCCTCCACTGGACGATGCCTCTTTCGTGAAGTATTTTGAGAAGGTGTCCATGAAGCCCTCTGGCTTCTTTAAGAACGTGCTGAATGAGGACGGCGATAAGGAGGAGGAGATCGAGAGGGTGCTGCCTGGAGCAGCCAATACATCTATGATCGTGCTGAAGAAGAAGGATCCATTCACCAGCTACGCCGCCGTGATCATCCCCCCTTCTGTGAGCAGGAATACCTTCTTTAAGGTGCAGTGCAACAATGACGAGTACAAGGAGGGCGCCAA GAACGGCGGCTATAAGGGCGTGATCCACCTGGACATCTCCCGCTCTGAGggtaccaagcac caccatcaccatcactaa

[0172] Pvs230Dl / Pvs230D7 IxTG Amino Acid Sequence (SEQ ID NO:66)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGQSVKGTGLCDFSKKSLIVPEPLTEESPPDIHCYSALKPLDTLYVKCPTEKAAYEVAKGKTGEEDEEEAKGIIALLEGDPADAGDPSAPPLDDASFVKYFEKVSMKPSGFFKNVLNEDGDKEEEIERVLPGAANTSMIVLKKKDPFTSYAAVIIPPSVSRN TFFKVQCNNDEYKEGAKNGGYKGVIHLDISRSEGTKHHHHHH

[0173] Pvs230Dl / Pvs230D7 4xTG Nucleic Acid Sequence (SEQ ID NO:67) accgggGTGCTGCCAGAGGCAGGAGCATCCGATGGCGTGTTTGACAAGGTGGATGAGG CCTTCGAGACCACAATCAAGGGCGACGGAAACGTGCTGCAGGCAAGCGATCCAGAG GTGGAGACATTTGCCAGCTCCAACACCAATAAGGAGTACGTGTGCGATTTCGTGAAGCACATCACCATGAAGGAGGCCTCCAAGAAGGTGGTCATCTGCGAGATGAAGATCCAGGAGCCACTGGTGAAGGTGAAGATCCTGTGCCCCACAAAGTACGCCGACGTGATCAAGTATGGCAGCATGGAGTTCTTTCCTAAGAAGGCCCCATACGTGACCCTGGTGAACGCCAATGAGGGCTCTGAGGAGAAGAAGGAGAGCCTGAAGGAGAAGCGGCTGGCCGCCCTGATCCACGGCGTGATCATCACCCCCGAGGTGAACGAGAAGGAGAACAATTTTGAGAAGGGCGTGATCGAGTTCGTGCTGCCACCCGTGCTGAAGGAGAAGAAGAAGCTGTACTATATCTGCGACAATGGCAAGTCTGCCGATGGCGTGAGCAACCACGGCGACAGAGGAGTGGCAGCAATCACAATCGAGCCCTACGGCCAGTCTGTGAAGGGCaccggaaca gggacgggcaccggtCTGTGCGATTTCTCCAAGAAGTCTCTGATCGTGCCCGAGCCTCTGACAGAGGAGTCTCCACCCGACATCCACTGCTACAGCGCCCTGAAGCCTCTGGATACACTGTATGTGAAGTGTCCAACCGAGAAGGCCGCCTACGAGGTGGCCAAGGGCAAGACCGGCGAGGAGGACGAGGAGGAGGCAAAGGGCATCATCGCCCTGCTGGAGGGCGATCCAGCAGACGCAGGCGATCCATCCGCCCCTCCACTGGACGATGCCTCTTTCGTGAAGTATTTTGAGAAGGTGTCCATGAAGCCCTCTGGCTTCTTTAAGAACGTGCTGAATGAGGACGGCGATAAGGAGGAGGAGATCGAGAGGGTGCTGCCTGGAGCAGCCAATACATCTATGATCGTGCTGAAGAAGAAGGATCCATTCACCAGCTACGCCGCCGTGATCATCCCCCCTTCTGTGAGCAGGAATACCTTCTTTAAGGTGCAGTGCAACAATGACGAGTACAAGGAGGGCGCCAAGAACGGCGGCTATAAGGGCGTGATCCACCTGGACATCTCCCGCTCTGAGggtaccaagcaccaccatcaccatcactaa

[0174] Pvs230Dl / Pvs230D7 4xTG Amino Acid Sequence (SEQ ID NO:68)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGOSVKGTGTGTGTGLCDF SKKSLIVPEPLTEESPPDIHC YS ALKPLDTLYVKCPTEKAAYEVAKGKTGEEDEEEAKGIIALLEGDPADAGDPSAPPLDDASFVKYFEKVSMKPSGFFKNVLNEDGDKEEEIERVLPGAANTSMIVLKKKDPFTSYAAVIIPPSVSRNTFFKVQCNNDEYKEGAKNGGYKGVIHLDISRSEGTKHHHHHH

[0175] Pvs230D8 Nucleic Acid Sequence (SEQ ID NO:69) accgggAAGAAGACAATCGGCTGCGACTTCACCGCCGAGACCTCCTCTATCTTCACCAAGGGCGTGCTGCTGCCAAGCGGACAGTCCAAGGAGTGTGAGGTGGAGGCCACCAAGAACGACATCTTCGGCGTGCGCTGCTCTAATGAGAGCACATTTGACCCTACCAACTGTTTCCACGAGATCTACGATAAGGCTGGCGGCAAGAAGAAGATCAAGGAGCTGATCCCAGACGTGAAGGTGTTTACACTGCCCAATAGCAAGACCAAGGTGGCCTACGGCAAGATCCCTCTGGATTATGTGAACAAGATCAATTTCAGCTGCTCCTGTACAAAGGCCGACGATTCTACAAAGGGCACCATCAAAGTGggtaccaagcaccaccatcaccatcactaa

[0176] Pvs230D8 Amino Acid Sequence (SEQ ID NO:70)TGKKTIGCDFTAETSSIFTKGVLLPSGQSKECEVEATKNDIFGVRCSNESTFDPTNCFHEIYDKAGGKKKIKELIPDVKVFTLPNSKTKVAYGKIPLDYVNKINFSCSCTKADDSTKGTIK VGTKHHHHHH

[0177] Pfs230Dl / Pvs230D8 IxTG Amino Acid Sequence (SEQ ID NO:71)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGKKTIGCDFTAETSSIFTKGVLLPSGQSKECEVEATKNDIFGVRCSNESTFDPTNCFHEIYDKAGGKKKIKELIPDVKVFTLPNSKTKVAYGKIPLDYVNKIN FSCSCTKADDSTKGTIKVGTKHHHHHH

[0178] Pvs230Dl / Pvs230D8 IxTG Nucleic Acid Sequence (SEQ ID NO:72) accgggGTGCTGCCAGAGGCAGGAGCATCCGATGGCGTGTTTGACAAGGTGGATGAGGCCTTCGAGACCACAATCAAGGGCGACGGAAACGTGCTGCAGGCAAGCGATCCAGAGGTGGAGACATTTGCCAGCTCCAACACCAATAAGGAGTACGTGTGCGATTTCGTGAAGCACATCACCATGAAGGAGGCCTCCAAGAAGGTGGTCATCTGCGAGATGAAGATCCAGGAGCCACTGGTGAAGGTGAAGATCCTGTGCCCCACAAAGTACGCCGACGTGATCAAGTATGGCAGCATGGAGTTCTTTCCTAAGAAGGCCCCATACGTGACCCTGGTGAACGCCAATGAGGGCTCTGAGGAGAAGAAGGAGAGCCTGAAGGAGAAGCGGCTGGCCGCCCTGATCCACGGCGTGATCATCACCCCCGAGGTGAACGAGAAGGAGAACAATTTTGAGAAGGGCGTGATCGAGTTCGTGCTGCCACCCGTGCTGAAGGAGAAGAAGAAGCTGTACTATATCTGCGACAATGGCAAGTCTGCCGATGGCGTGAGCAACCACGGCGACAGAGGAGTGGCAGCAATCACAATCGAGCCCTACGGCCAGTCTGTGAAGGGCaccggaAAGAAGACAATCGGCTGCGACTTCACCGCCGAGACCTCCTCTATCTTCACCAAGGGCGTGCTGCTGCCAAGCGGACAGTCCAAGGAGTGTGAGGTGGAGGCCACCAAGAACGACATCTTCGGCGTGCGCTGCTCTAATGAGAGCACATTTGACCCTACCAACTGTTTCCACGAGATCTACGATAAGGCTGGCGGCAAGAAGAAGATCAAGGAGCTGATCCCAGACGTGAAGGTGTTTACACTGCCCAATAGCAAGACCAAGGTGGCCTACGGCAAGATCCCTCTGGATTATGTGAACAAGATCAATTTCAGCTGCTCCTGTACAAAGGCCGACGATTCTACAAAGGGCACCATCAAAGTGggtaccaagcaccaccatcaccatcactaa

[0179] Pfs230Dl / Pvs230D8 IxTG Nucleic Acid Sequence (SEQ ID NO:73) accggtTCCGTGCTGCAGTCTGGCGCCCTGCCTAGCGTGGGAGTGGACGAGCTGGATAAGATCGACCTGTCCTACGAGACCACAGAGTCTGGCGACACAGCCGTGTCTGAGGATAGCTATGACAAGTACGCCAGCAACAATACCAACAAGGAGTACGTGTGCGATTTCACCGACCAGCTGAAGCCTACAGAGTCCGGCCCAAAGGTGAAGAAGTGCGAGGTGAAGGTGAACGAGCCTCTGATCAAGGTGAAGATCATCTGTCCACTGAAGGGCTCTGTGGAGAAGCTGTATGATAATATCGAGTACGTGCCCAAGAAGAGCCCTTACGTGGTGCTGACCAAGGAGGAGACAAAGCTGAAGGAGAAGCTGCTGAGCAAGCTGATCTATGGCCTGCTGATCTCCCCCACCGTGAACGAGAAGGAGAACAACTTCAAGGAGGGCGTGATCGAGTTCACACTGCCCCCTGTGGTGCACAAGGCCACCGTGTTCTACTTTATCTGCGACAATAGCAAGACCGAGGATGACAATAAGAAGGGCAACAGGGGCATCGTGGAGGTGTATGTGGAGCCATACGGCAACAAGATCAATGGCaccggtAAGAAGACAATCGGCTGCGACTTCACCGCCGAGACCTCCTCTATCTTCACCAAGGGCGTGCTGCTGCCAAGCGGACAGTCCAAGGAGTGTGAGGTGGAGGCCACCAAGAACGACATCTTCGGCGTGCGCTGCTCTAATGAGAGCACATTTGACCCTACCAACTGTTTCCACGAGATCTACGATAAGGCTGGCGGCAAGAAGAAGATCAAGGAGCTGATCCCAGACGTGAAGGTGTTTACACTGCCCAATAGCAAGACCAAGGTGGCCTACGGCAAGATCCCTCTGGATTATGTGAACAAGATCAATTTCAGCTGCTCCTGTACAAAGGCCGACGATTCTACAAAGGGCACCATCAAAGTGggtacca agcaccaccatcaccatcactaa

[0180] Pvs230Dl / Pvs230D8 lxTG Amino Acid Sequence (SEQ ID NO:74)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGQSVKGTGKKTIGCDFTAETSSIFTKGVLLPSGQSKECEVEATKNDIFGVRCSNESTFDPTNCFHEIYDKAGGKKKIKELIPDVKVFTLPNSKTKVAYGKIPLDYVNKINF SCSCTKADD STKGTIKVGTKHHHHHH

[0181] Pvs230Dl / Pvs230D8 4xTG Nucleic Acid Sequence (SEQ ID NO:75) accgggGTGCTGCCAGAGGCAGGAGCATCCGATGGCGTGTTTGACAAGGTGGATGAGGCCTTCGAGACCACAATCAAGGGCGACGGAAACGTGCTGCAGGCAAGCGATCCAGAGGTGGAGACATTTGCCAGCTCCAACACCAATAAGGAGTACGTGTGCGATTTCGTGAAGCACATCACCATGAAGGAGGCCTCCAAGAAGGTGGTCATCTGCGAGATGAAGATCCAGGAGCCACTGGTGAAGGTGAAGATCCTGTGCCCCACAAAGTACGCCGACGTGATCAAGTATGGCAGCATGGAGTTCTTTCCTAAGAAGGCCCCATACGTGACCCTGGTGAACGCCAATGAGGGCTCTGAGGAGAAGAAGGAGAGCCTGAAGGAGAAGCGGCTGGCCGCCCTGATCCACGGCGTGATCATCACCCCCGAGGTGAACGAGAAGGAGAACAATTTTGAGAAGGGCGTGATCGAGTTCGTGCTGCCACCCGTGCTGAAGGAGAAGAAGAAGCTGTACTATATCTGCGACAATGGCAAGTCTGCCGATGGCGTGAGCAACCACGGCGACAGAGGAGTGGCAGCAATCACAATCGAGCCCTACGGCCAGTCTGTGAAGGGCaccggaaca gggacgggcaccggtAAGAAGACAATCGGCTGCGACTTCACCGCCGAGACCTCCTCTATCTTCACCAAGGGCGTGCTGCTGCCAAGCGGACAGTCCAAGGAGTGTGAGGTGGAGGCCACCAAGAACGACATCTTCGGCGTGCGCTGCTCTAATGAGAGCACATTTGACCCTACCAACTGTTTCCACGAGATCTACGATAAGGCTGGCGGCAAGAAGAAGATCAAGGAGCTGATCCCAGACGTGAAGGTGTTTACACTGCCCAATAGCAAGACCAAGGTGGCCTACGGCAAGATCCCTCTGGATTATGTGAACAAGATCAATTTCAGCTGCTCCTGTACAAAGGCCGACGATTCTACAAAGGGCACCATCAAAGTGggtaccaagcaccaccatcaccatcactaa

[0182] Pvs230Dl / Pvs230D8 4xTG Amino Acid Sequence (SEQ ID NO:76)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHI TMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGS EEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGQSVKGTGTGTGTGKKTIGCDFTAETSSIFTKGVLLPSGQS KECEVEATKNDIFGVRCSNESTFDPTNCFHEIYDKAGGKKKIKELIPDVKVFTLPNSKTK VAYGKIPLDYVNKINFSCSCTKADDSTKGTIKVGTKHHHHHH

[0183] Pfs230D14 Nucleic Acid Sequence (SEQ ID NO:77)ACCGGTTCCAAGAAGCCCCTGCCTAACGACGATGATATCTGCAACGTGACCATCGGCAACAACACATTTTCTGGCTTCGCCTGCCTGAGCCACTTCGAGCTGAAGCCAAACAATTGCTTCAGCAGCGTGTACGACTACAACGAGGCCAACAAGGTCAAGAAACTGTTCGACCTGTCTACAAAGGTGGAACTGGACCACATCAAACAGAACACCAGCGGATACACC CTGAGCTACATCATTTTCAACAAGGAAAGCACCAAGCTCAAGTTCTCCTGCACCTGT TCTAGCAACTACAGCAATTATACAATCCGGATCACCTTTGATCCTAATTACATCATCCCCGAGCCTCAGAGCAGAGCCATCATCAAGTACGTGGACCTGCAGGACAAGAACTTCGCTAAATACCTGAGAAAGCTGGGTACCAAGCACCACCATCACCATCACTAA

[0184] Pfs230D14 Amino Acid Sequence (SEQ ID NO:78)TG SKKPLPNDDDICNVTIGNNTF SGF ACL SHFELKPNNCF S S VYD YNEANKVKKLFDL ST KVELDHIKQNTSGYTLSYIIFNKESTKLKFSCTCSSNYSNYTIRITFDPNYIIPEPQSRAIIKY VDLQDI<NFAI<YLRI<LGTKHHHHHH

[0185] Pfs230Dl / Pfs230D14 lx TG Nucleic Acid Sequence (SEQ ID NO:79)ACCGGTAGCGTGCTGCAGAGCGGCGCCCTGCCTAGCGTGGGCGTCGACGAGCTGGACAAGATCGACCTGTCTTACGAGACAACCGAGTCCGGCGACACCGCCGTGTCCGAGGACAGCTACGACAAGTACGCCTCTAATAACACAAACAAGGAATACGTGTGCGACTTTACCGACCAACTGAAACCCACCGAGAGCGGACCTAAGGTGAAGAAATGCGAGGTGA AGGTTAACGAGCCTCTGATCAAGGTCAAGATCATTTGTCCTCTGAAGGGAAGCGTGG AAAAACTGTACGATAACATCGAGTACGTGCCAAAGAAGTCCCCTTATGTGGTCCTGACCAAGGAGGAAACAAAGCTGAAGGAAAAGCTGCTGAGCAAGCTCATCTACGGCCTGCTGATCAGCCCTACCGTGAACGAGAAGGAAAACAATTTTAAGGAGGGCGTGATCGA GTTCACCCTGCCCCCCGTGGTGCACAAGGCTACAGTGTTCTACTTCATCTGTGATAAT AGCAAAACCGAGGATGATAACAAGAAGGGTAATAGAGGCATCGTGGAAGTGTACG TGGAACCATACGGCAACAAGATTAACGGCACAGGCACCGGCACCGGAACAGGCAACCAGGGCTACAAGGAGATCCACGGCTGCGACTTCACAGGCAAGTACAGCCACCTGTTCACCTACTCCAAGAAGCCCCTGCCTAACGACGATGATATCTGCAACGTGACCATCG GCAACAACACATTTTCTGGCTTCGCCTGCCTGAGCCACTTCGAGCTGAAGCCAAACA ATTGCTTCAGCAGCGTGTACGACTACAACGAGGCCAACAAGGTCAAGAAACTGTTC GACCTGTCTACAAAGGTGGAACTGGACCACATCAAACAGAACACCAGCGGATACACCCTGAGCTACATCATTTTCAACAAGGAAAGCACCAAGCTCAAGTTCTCCTGCACCTG TTCTAGCAACTACAGCAATTATACAATCCGGATCACCTTTGATCCTAATTACATCATC CCCGAGCCTCAGAGCAGAGCCATCATCAAGTACGTGGACCTGCAGGACAAGAACTT CGCTAAATACCTGAGAAAGCTGGGTACCAAGCACCACCATCACCATCACTAA

[0186] Pfs230Dl / Pfs230D14 lx TG Amino Acid Sequence (SEQ ID NO:80)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKL KEKLL SKLIYGLLISPT VNEKENNFKEGVIEFTLPP VVHK AT VF YFICDN SKTEDDNKKGNRGIVEVYVEPYGNKINGTGNQGYKEIHGCDFTGKYSHLFTYSKKPLPNDDDICNVTIGN NTF SGF ACL SHFELKPNNCF S S V YD YNEANKVKKLFDLSTKVELDHIKQNT SGYTL S YII FNKESTKLKFSCTCSSNYSNYTIRITFDPNYIIPEPQSRAIIKYVDLQDKNFAKYLRKLGTK HHHHHH

[0187] Pfs230Dl / Pfs230D14 4x TG Nucleic Acid Sequence (SEQ ID NO:81)ACCGGTAGCGTGCTGCAGAGCGGCGCCCTGCCTAGCGTGGGCGTCGACGAGCTGGA CAAGATCGACCTGTCTTACGAGACAACCGAGTCCGGCGACACCGCCGTGTCCGAGGACAGCTACGACAAGTACGCCTCTAATAACACAAACAAGGAATACGTGTGCGACTTT ACCGACCAACTGAAACCCACCGAGAGCGGACCTAAGGTGAAGAAATGCGAGGTGA AGGTTAACGAGCCTCTGATCAAGGTCAAGATCATTTGTCCTCTGAAGGGAAGCGTGGAAAAACTGTACGATAACATCGAGTACGTGCCAAAGAAGTCCCCTTATGTGGTCCTGA CCAAGGAGGAAACAAAGCTGAAGGAAAAGCTGCTGAGCAAGCTCATCTACGGCCTG CTGATCAGCCCTACCGTGAACGAGAAGGAAAACAATTTTAAGGAGGGCGTGATCGA GTTCACCCTGCCCCCCGTGGTGCACAAGGCTACAGTGTTCTACTTCATCTGTGATAATAGCAAAACCGAGGATGATAACAAGAAGGGTAATAGAGGCATCGTGGAAGTGTACGTGGAACCATACGGCAACAAGATTAACGGCACAGGCAACCAGGGCTACAAGGAGATCCACGGCTGCGACTTCACAGGCAAGTACAGCCACCTGTTCACCTACTCCAAGAAGCCCCTGCCTAACGACGATGATATCTGCAACGTGACCATCGGCAACAACACATTTTCTGGCTTCGCCTGCCTGAGCCACTTCGAGCTGAAGCCAAACAATTGCTTCAGCAGCGTGTACGACTACAACGAGGCCAACAAGGTCAAGAAACTGTTCGACCTGTCTACAAAGGTGGAACTGGACCACATCAAACAGAACACCAGCGGATACACCCTGAGCTACATCATTTTCAACAAGGAAAGCACCAAGCTCAAGTTCTCCTGCACCTGTTCTAGCAACTACAGCAA TTATACAATCCGGATCACCTTTGATCCTAATTACATCATCCCCGAGCCTCAGAGCAGAGCCATCATCAAGTACGTGGACCTGCAGGACAAGAACTTCGCTAAATACCTGAGAA AGCTGGGTACCAAGCACCACCATCACCATCACTAA

[0188] Pfs230Dl / Pfs230D14 4x TG Amino Acid Sequence (SEQ ID NO:82)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGTGNQGYKEIHGCDFTGKYSHLFTYSKKPLPNDDDIC NVTIGNNTF SGF ACLSHFELKPNNCF S S VYDYNEANKVKKLFDLSTKVELDHIKQNTSG YTLSYIIFNKESTKLKFSCTCSSNYSNYTIRITFDPNYIIPEPQSRAIIKYVDLQDKNFAKYL RKLGTKHHHHHH

[0189] Pfs230D7 Nucleic Acid Sequence (SEQ ID NO:83) accggtAACAGGCACGTGTGCGACTTCTCTAAGAATAACCTGATCGTGCCTGAGAGCCT GAAGAAGAAGGAAGAGCTGGGCGGCAATCCAGTGAACATCCACTGCTATGCCCTGC TGAAGCCACTGGACACCCTGTACGTGAAGTGTCCCACATCTAAGGATAACTATGAG GCCGCCAAAGTGAATATCAGCGAGAACGACAATGAGTACGAGCTGCAGGTCATCTCCCTGATCGAGAAGCGGTTCCACAACTTCGAGACCCTGGAGAGCAAGAAGCCCGGCAACGGCGATGTGGTGGTGCACAATGGCGTGGTGGACACCGGCCCTGTGCTGGATAACTCCACATTTGAGAAGTACTTCAAGAACATCAAGATCAAGCCTGACAAGTTCTTTGAGAAAGTGATCAACGAGTATGATGACACCGAGGAGGAGAAGGATCTGGAGTCCATCCTGCCCGGCGCCATCGTGTCTCCTATGAAGGTGCTGAAGAAGAAGGACCCCTTCACCAGCTACGCAGCCTTCGTGGTGCCCCCTATCGTGCCAAAGGACCTGCACTTTAAGGTGGAGTGTAATAACACCGAGTATAAGGATGAGAACCAGTATATCTCTGGCTACAATGGC ATCATCCACATCGACATCAGCAACTCCggtaccaagcaccaccatcaccatcactaa

[0190] Pfs230D7 Amino Acid Sequence (SEQ ID NO:84)TGNRHVCDFSKNNLIVPESLKKKEELGGNPVNIHCYALLKPLDTLYVKCPTSKDNYEAAKVNISENDNEYELQVISLIEKRFHNFETLESKKPGNGDVVVHNGVVDTGPVLDNSTFEKYFKNIKIKPDKFFEKVINEYDDTEEEKDLESILPGAIVSPMKVLKKKDPFTSYAAFVVPPI VPKDLHFKVECNNTEYKDENQYISGYNGIIHIDISNSGTKHHHHHH

[0191] Pfs230D9 Nucleic Acid Sequence (SEQ ID NO:85) accggtAATGTGCACCTGTGCAATTTCTTTGACAACCCCGAGCTGACCTTCGATAACAA TAAGATCGTGCTGTGCAAGATCGACGCCGAGCTGTTTAGCGAAGTGATCATCCAGCT GCCTATCTTCGGCACCAAGAACGTGGAGGAGGGCGTGCAGAATGAGGAGTACAAGAAGTTTTCTCTGAAGCCAAGCCTGGTGTTCGATGACAACAATAACGACATCAAAGTGATCGGCAAGGAGAAGAACGAGGTGTCCATCTCTCTGGCCCTGAAGGGCGTGTATGGCAACAGGATCTTTACCTTCGATAAGAATGGCAAGAAGGGCGAGGGCATCTCCTTCTTTATCCCACCCATCAAGCAGGACACAGATCTGAAGTTTATCATCAACGAGACCATCGA CAACTCTAATATCAAGCAGAGGGGCCTGATCTACATCTTCGTGCGCAAGAATggtaccaa gcaccaccatcaccatcactaa

[0192] Pfs230D9 Amino Acid Sequence (SEQ ID NO:86)TGNVHLCNFFDNPELTFDNNKIVLCKIDAELFSEVIIQLPIFGTKNVEEGVQNEEYKKFSL KPSLVFDDNNNDIKVIGKEKNEVSISLALKGVYGNRIFTFDKNGKKGEGISFFIPPIKQDT DLKFIINETIDNSNIKQRGLIYIFVRKNGTKHHHHHH

[0193] Pfs230D2-3 Nucleic Acid Sequence (SEQ ID NO:87) accggtTGTGCCTTCCTGGACGAGGATGAGGAGGAGGAGAAGTACGGCAACCAGATCGAGGAGGACGAGCACAATGAGAAGATCAAGATGAAGACCTTCTTTACACAGAACATCTACAAGAAGAACAATATCTATCCCTGCTACATGAAGCTGTATTCCGGCGATATCGGCGGCATCCTGTTTCCAAAGAATATCAAGTCTACCACATGTTTCGAGGAGATGATCCCCTACAACAAGGAGATCAAGTGGAACAAGGAGAATAAGAGCCTGGGCAATCTGGTGAACAATTCCGTGGTGTACAATAAGGAGATGAACGCCAAGTACTTCAACGTGCAGTACGTGCACATCCCCACCTCTTATAAGGACACACTGAATCTGTTCTGCTCTATCATCCTGAAGGAGGAGGAGAGCAACCTGATCAGCACATCCTATCTGGTGTACGTGAGCATCAATGAGGAGCTGAACTTTAGCCTGTTCGACTTTTACGAGTCCTTCGTGCCTATCAAGAAGACCATCCAGGTGGCCCAGAAGAACGTGAACAATAAGGAGCACGATTATACCTGTGACTTCACCGACAAGCTGGATAAGACCGTGCCATCCACAGCCAATGGCAAGAAGCTGTTCATCTGCCGGAAGCACCTGAAGGAGTTTGACACCTTCACACTGAAGTGTAACGTGAATAAGACACAGTACCCAAACATCGAGATCTTCCCCAAGACCCTGAAGGATAAGAAGGAGGTGCTGAAGCTGGACCTGGATATCCAGTATCAGATGTTTAGCAAGTTCTTTAAGTTCAACACCCAGAATGCCAAGTATCTGAATCTGTACCCTTACTATCTGATCTTCCCCTTCAACCACATCGGCAAGAAGGAGCTGAAGAACAATCCAACCTATAAGAACCACAAGGACGTGAAGTACTTTGAGCAGTCCTCTGTGCTGTCTCCACTGAGCTCCGCCGACAGCCTGGGCAAGCTGCTGAATTTCCTGGATACCCAGGAGACAGTGTGCCTGACCGAGAAGATCAGATACCTGAATCTGAGCATCAACGAGCTGGGCTCCGACAACAATACCTTTTCTGTGACATTCCAGGTGCCACCCTATATCGATATCAAGGAGCCTTTCTACTTTATGTTCGGCTGCAACAATAACAAGGGCGAGGGCAACATCGGCATCGTGGAGCTGCTGATCAGCAAGggtaccaagcaccaccatcaccatcactaa

[0194] Pfs230D2-3 Amino Acid Sequence (SEQ ID NO:88)TGCAFLDEDEEEEKYGNQIEEDEHNEKIKMKTFFTQNIYKKNNIYPCYMKLYSGDIGGILFPKNIKSTTCFEEMIPYNKEIKWNKENKSLGNLVNNSVVYNKEMNAKYFNVQYVHIPTSYKDTLNLFCSIILKEEESNLISTSYLVYVSINEELNFSLFDFYESFVPIKKTIQVAQKNVNNKEHDYTCDFTDKLDKTVPSTANGKKLFICRKHLKEFDTFTLKCNVNKTQYPNIEIFPKTLKDKKEVLKLDLDIQYQMFSKFFKFNTQNAKYLNLYPYYLIFPFNHIGKKELKNNPTYKNHKD VK YFEQ S S VLSPLS S AD SLGKLLNFLDTQETVCLTEKIRYLNL SINELGSDNNTF S VTFQVPPYIDIKEPFYFMFGCNNNKGEGNIGIVELLISKGTKHHHHHH

[0195] Pfs230D2 Nucleic Acid Sequence (SEQ ID NO:89) accggtTGTGCCTTCCTGGACGAGGATGAGGAGGAGGAGAAGTACGGCAACCAGATCGAGGAGGACGAGCACAATGAGAAGATCAAGATGAAGACCTTCTTTACACAGAACATCTACAAGAAGAACAATATCTATCCCTGCTACATGAAGCTGTATTCCGGCGATATCGGCGGCATCCTGTTTCCAAAGAATATCAAGTCTACCACATGTTTCGAGGAGATGATCCCCTACAACAAGGAGATCAAGTGGAACAAGGAGAATAAGAGCCTGGGCAATCTGGTGAACAATTCCGTGGTGTACAATAAGGAGATGAACGCCAAGTACTTCAACGTGCAGTACGTGCACATCCCCACCTCTTATAAGGACACACTGAATCTGTTCTGCTCTATCATCCTGAAGGAGGAGGAGAGCAACCTGATCAGCACATCCTATCTGGTGTACGTGAGCATCAATGAGGAGCTGAACTTTAGCCTGTTCGACTTTTACGAGTCCTTCGTGCCTATCAAGAAGACCATCCAGGTGGCCggtaccaagcaccaccatcaccatcactaa

[0196] Pfs230D2 Amino Acid Sequence (SEQ ID NO:90)TGCAFLDEDEEEEKYGNQIEEDEHNEKIKMKTFFTQNIYKKNNIYPCYMKLYSGDIGGILFPKNIKSTTCFEEMIPYNKEIKWNKENKSLGNLVNNSVVYNKEMNAKYFNVQYVHIPTSYKDTLNLFCSIILKEEESNLISTSYLVYVSINEELNFSLFDFYESFVPIKKTIQVAGTKHHHI II II I

[0197] Pfs48D3 Nucleic Acid Sequence (SEQ ID NO:91) accggtGAGAAGAAAGTGATCCACGGCTGCAACTTCTCTTCCAATGTGAGCTCTAAGCATACCTTTACAGACAGCCTGGATATCTCTCTGGTGGACGATTCCGCTCACATCAGCTGTAACGTGCATCTGAGCGAGCCTAAGTATAACCATCTGGTGGGCCTGAATTGCCCAGGCGACATCATCCCCGATTGTTTCTTTCAGGTGTACCAGCCTGAGTCTGAGGAGCTGGAGCCATCCAACATCGTGTATCTGGATTCTCAGATCAATATCGGCGACATCGAGTACTATGAGGATGCTGAGGGCGACGATAAGATCAAGCTGTTCGGCATCGTGGGCAGCATCCCAAAGACCACATCTTTCACCTGTATCTGTAAAAAGGACAAGAAATCAGCCTATATGACTGTGACCATTGACTCAggtaccaagcaccaccatcaccatcactaa

[0198] Pfs48D3 Amino Acid Sequence (SEQ ID NO:92)TGEKK V1HGC NFS SNVS SKHTFTDSLDISLVDD S AHISCNVHLSEPKYNHLVGLNCPGDII PDCFFQVYQPESEELEPSNIVYLDSQINIGDIEYYEDAEGDDKIKLFGIVGSIPKTTSFTCIC KKDKKSAYMTVTIDSGTKHHHHHH

[0199] Pfs230Dl / Pvs230D7 4xTG Amino Acid Sequence (SEQ ID NO:93)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGTGLCDFSKKSLIVPEPLTEESPPDIHCYSALKPLDTLYVKCPTEKAAYEVAKGKTGEEDEEEAKGIIALLEGDPADAGDPSAPPLDDASFVKYFEKVSMKPSGFFKNVLNEDGDKEEEIERVLPGAANTSMIVLKKKDPFTSYAAVIIPPSVSRNTFFKVQCNNDEYKEGAKNGGYKGV1HLDISRSEGTKHHHHHH

[0200] Pvs230Dl / Pvs230D7 lxTG Amino Acid Sequence (SEQ ID NO:94)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHI TMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSA DGVSNHGDRGVAAITIEPYGQSVKGTGLCDFSKKSLIVPEPLTEESPPDIHCYSALKPLDT LYVKCPTEKAAYEVAKGKTGEEDEEEAKGIIALLEGDPADAGDPSAPPLDDASFVKYFEKVSMKPSGFFKNVLNEDGDKEEEIERVLPGAANTSMIVLKKKDPFTSYAAVIIPPSVSRN TFFKVQCNNDEYKEGAKNGGYKGVIHLDISRSEGTKHHHHHH

[0201] Pvs230Dl / Pvs230D7 4xTG Amino Acid Sequence (SEQ ID NO:95)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHI TMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGS EEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSA DGVSNHGDRGVAAITIEP YGQ S VKGTGTGTGTGLCDF SKKSLIVPEPLTEESPPDIHC YS A LKPLDTLYVKCPTEKAAYEVAKGKTGEEDEEEAKGIIALLEGDPADAGDPSAPPLDDASFVKYFEKVSMKPSGFFKNVLNEDGDKEEEIERVLPGAANTSMIVLKKKDPFTSYAAVIIPPSVSRNTFFKVQCNNDEYKEGAKNGGYKGVIHLDISRSEGTKHHHHHH

[0202] Pfs230Dl / Pvs230D8 4xTG Amino Acid Sequence (SEQ ID NO:96)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLL SKLIYGLLISPT VNEKENNFKEGVIEFTLPP VVHKAT VF YFICDN SKTEDDNKKGNRGIVEVYVEPYGNKINGTGTGTGTGKKTIGCDFTAETSSIFTKGVLLPSGQSKECEVEATKNDIFGVRCSNESTFDPTNCFHEIYDKAGGKKKIKELIPDVKVFTLPNSKTKVAYGKIPLDYVNKINFSCSCTKADDSTKGTIKVGTKHHHHHH

[0203] Pvs230Dl / Pvs230D8 lxTG Amino Acid Sequence (SEQ ID NO:97)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGQSVKGTGKKTIGCDFTAETSSIFTKGVLLPSGQSKECEVEATKNDIFGVRCSNESTFDPTNCFHEIYDKAGGKKKIKELIPDVKVFTLPNSKTKVAYGKIPLDYVNKINF SCSCTKADD STKGTIKVGTKHHHHHH

[0204] Pvs230Dl / Pvs230D8 4xTG Amino Acid Sequence (SEQ ID NO:98)TGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETFASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGQSVKGTGTGTGTGKKTIGCDFTAETSSIFTKGVLLPSGQSKECEVEATKNDIFGVRCSNESTFDPTNCFHEIYDKAGGKKKIKELIPDVKVFTLPNSKTKVAYGKIPLDYVNKINFSCSCTKADDSTKGTIKVGTKHHHHHH

[0205] Pfs230Dl / Pvs230Dl lxTG Amino Acid Sequence (SEQ ID NO:99)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLL SKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVF YFICDN SKTEDDNKKGNRGIVEVYVEPYGNKINGTGVLPEAGASDGVFDKVDEAFETTIKGDGNVLQASDPEVETF ASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKYGSMEF FPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIEFVLPPV LKEKKKLYYICDNGKSADGVSNHGDRGVAAITIEPYGQSVKGTKHHHHHH

[0206] Pfs230Dl / Pvs230Dl 4xTG Amino Acid Sequence (SEQ ID NO: 100)TGSVLQSGALPSVGVDELDKIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQ LKPTESGPKVKKCEVKVNEPLIKVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKL KEKLLSKLIYGLLISPTVNEKENNFKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGN RGIVEVYVEPYGNKINGTGTGTGTGVLPEAGASDGVFDKVDEAFETTIKGDGNVLOASD PEVETFASSNTNKEYVCDFVKHITMKEASKKVVICEMKIQEPLVKVKILCPTKYADVIKY GSMEFFPKKAPYVTLVNANEGSEEKKESLKEKRLAALIHGVIITPEVNEKENNFEKGVIE FVLPPVLKEKKKLYYICDNGKSADGVSNHGDRGVAAIT1EPYGQSVKGTKHELFLFLHH

[0207] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

[0208] The use of the terms “a” and “an” and “the” and “at least one” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term “at least one” followed by a list of one or more items (for example, “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methodsdescribed herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0209] Preferred embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

CLAIM(S):

1. A protein, comprising, from amino- to carboxy termini: (a) a first polypeptide domain derived from the Pfs230 domain 1 (Pfs230Dl) of Plasmodium falciparum or the Pvs230 domain 1 (Pvs230Dl) of Plasmodium vivax, (b) an optional linker domain, and (c) a second polypeptide domain, wherein the fusion protein is not the native Pfs230 or Pvs230 protein.

2. The protein of claim 1, wherein the first polypeptide domain comprises an amino acid sequence, which sequence comprises, consists essentially of, or consists of SEQ ID NO: 1 or SEQ ID NO:36, wherein the N-terminal TG dipeptide is either present or absent.

3. The protein of claim 1 or 2, wherein the amino acid sequence of the first polypeptide domain consists essentially of SEQ ID NO: 1 or SEQ ID NO:36, wherein the N-terminal TG dipeptide is either present or absent.

4. The protein of any one of claims 1-3, wherein the amino acid sequence of the first polypeptide domain is at least 90% identical to SEQ ID NO: 1, wherein the N-terminal TG dipeptide is either present or absent.

5. The protein of any one of claims 1-3, wherein the amino acid sequence of the first polypeptide domain is at least 90% identical to SEQ ID NO:36, wherein the N-terminal TG dipeptide is either present or absent.

6. The protein of any one of claims 1-5, wherein the linker domain consists of 0 amino acids (i.e., is absent).

7. The protein of any one of claims 1-6, wherein the linker domain consists of between 1 and 20 amino acids.

8. The protein of claim 7, wherein the linker domain does not form a secondary structure consisting of either an a-helix or a P-sheet.

9. The protein of any one of claims 1-8, wherein the second polypeptide domain comprises, consists essentially of, or consists of a 6-Cys polypeptide.

10. The protein of claim 9, wherein the second polypeptide domain comprises, consists essentially of, or consists of a polypeptide derived from a species within the aconoidasidan (hematozoan) clade of Apicocomplexa.

11. The protein of claim 9, wherein the second polypeptide domain comprises, consists essentially of, or consists of Pfs48 / 45D3, Pfs230D2, Pfs230D3, Pfs230D4, Pfs230D7, Pfs230D9, Pvs230Dl, Pvs48D3, Pvs230D7, Pvs230D8, or orthologs thereof.

12. The protein of any one of claims 1-5, which comprises an amino acid sequence comprising, consisting essentially of, or comprising one of SEQ ID NOs:2-16, 18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100.

13. The protein of any one of claims 1-5, which comprises an amino acid sequence at least 95% identical to one of SEQ ID NOs:2-16, 18, 40, 44, 48, 52, 56, 60, 64, 66, 68, 71, 74, 76, 80, 82, or 93-100.

14. A nucleic acid comprising an open reading frame (ORF) encoding the protein of any one of claims 1-13.

15. A gene-transfer vector comprising the nucleic acid of claim 14.

16. A cell comprising the nucleic acid of claim 14 or the gene-transfer vector of claim 15.

17. A population of cells comprising the cell of claim 16.

18. A pharmaceutical composition comprising the protein of any one of claims 1-13 and a pharmaceutically-acceptable carrier.

19. The pharmaceutical composition of claim 18, comprising an immune adjuvant.

20. The pharmaceutical composition of claim 19, wherein the immune adjuvant comprises alum, AS01 or MATRIX M.

21. A method for vaccinating a subject against a pathogen, comprising administering the protein of any one of claims 1-13 or the pharmaceutical composition of any one of claims 18-20 in an immunogenically-effective amount and in a manner and at a location sufficient to elicit an immune response in the subject to the protein, whereby the subject acquires immunity to a pathogen.

22. Use of the protein of any one of claims 1-13 or the pharmaceutical composition of any one of claims 18-20 as a vaccine to vaccinate a subject against a pathogen.

23. The method of claim 21 or the use of claim 22, wherein the pathogen is a parasite.

24. The method or use of any one of claims 21-23, wherein the pathogen is a species of Plasmodium.

25. The method or use of any one of claims 21-24, whereby the subject acquires immunity protective against malaria.

26. The method or use of any one of claims 21-25, wherein the protein or composition is administered by intravenous injection.

27. The method or use of any one of claims 21-26, wherein the patient is treated with an additional pharmaceutical agent or vaccine.

28. The method or use of claim 27, wherein the additional pharmaceutical agent or vaccine comprises alum, AS01 or MATRIX M.

29. The method or use of any one of claims 21-28, wherein the subject is human.

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