Application of plant JAZ protein and derived polypeptides in preventing and treating tumors in humans and animals

Abstract

The present invention discloses the application of plant JAZ proteins and derived polypeptides in the prevention and treatment of tumors in humans and animals. The present invention provides the application of a plant JAZ protein or its derived polypeptide or its modified product or its related biological material in the preparation of a product for treating and / or preventing tumors in humans or animals, inhibiting the occurrence and development of tumors in humans or animals, inhibiting the proliferation of tumor cells in humans or animals, and promoting the apoptosis of tumor cells in humans or animals. Cotton JAZ protein has a significant inhibitory effect on a variety of human tumor cell lines and has a significant inhibitory effect on the occurrence and development of tumors in mice in vivo. JAZ protein has a strong interaction with the human proto-oncogene MYCs. JAZ protein can target various types of human and animal tumors and can be developed into a targeted drug for human and animal tumors. The present invention provides a new drug or adjuvant drug for the treatment of various human and animal tumors.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and particularly to the application of plant JAZ proteins and derived polypeptides in the prevention and treatment of tumors in humans and animals. Background Art

[0002] Plant callus culture is one of the key technical links in the breeding process of gene editing technology. Plant callus is a physiological tumor formed during the wound healing process after a plant is injured under adversity. It is a mass of amorphous parenchyma cells that have lost their original structure and function and regained the ability to divide, and can proliferate indefinitely. Plants can produce "tumors" to heal wounds, and the rapid proliferation of cells will be inhibited after the wound is healed, without the phenomenon of malignant proliferation. During the development of plant callus (plant tumor), the JA signaling pathway is the key signaling pathway that regulates the development of plant "tumors" and promotes wound healing. The plant-specific JAZ protein is the only negative regulator of the jasmonic acid signaling pathway and is the key factor that prevents the development of "plant tumors". JAZ protein is a plant-specific protein and is abundant in cotton. It has been found that overexpression of cotton JAZ can inhibit the growth of cotton callus during tissue culture, and JA inhibitors can also inhibit the growth of cotton callus. Moreover, the inhibition of JAZ on JA signaling is achieved by inhibiting the transcriptional factor function of MYC.

[0003] "Plant tumors" are similar to human tumors, such as similar morphology, both are induced by external stimuli, have fast cell division, rapid proliferation, and are both regulated by the transcriptional factor MYC. The growth of "plant tumors" is strictly regulated by the JA signal and cannot develop malignantly. Different from the controllable "plant tumors", human tumors often deteriorate and develop into cancer due to uncontrollability. Cancer seriously affects human health, and it is urgent to explore the mechanism of cancer occurrence and develop cancer treatment strategies. The activation of the proto-oncogene MYC is related to the occurrence and development of various types of cancers and is the most widely studied oncogene. Given the key role of MYC in the occurrence and development of cancer, screening drugs that target and inhibit MYC activity is an important strategy for treating malignant tumors. Currently, for drugs targeting MYC, the most studied is to inhibit the interaction between MYC / MAX, such as small molecule inhibitors Mycro 3, SI-3716, MYCMI-6, sAJM589, etc., polypeptide drug OmoMYC, natural products Stauprimide, Lusianthridin, etc. Since MYC and MAX can form a complex structure composed of four helical bundles, the interaction between the targets is complex, and it is difficult to develop drugs based on this structure. Therefore, it is of great significance to find natural proteins that are easy to produce and can effectively inhibit MYC. Summary of the Invention

[0004] The object of the present invention is to provide a new drug for tumor prevention and treatment, namely JAZ protein, JAZ homologous protein and their derived polypeptides, etc., which have important pharmaceutical application value in tumor prevention and treatment.

[0005] The technical problem to be solved by the present invention is to overcome the problems such as the difficulty in drug research and development based on the complex interaction between the proto-oncogene MYCs (c-Myc, N-Myc, L-Myc) targets in human and animal tumors, aiming to explore drugs for preventing and treating tumors against the harm of tumors to humans and animals for a long time.

[0006] The present invention provides a mechanism and natural products for multi-target anti-tumor, which solves the problems of single target, drug resistance and low efficiency of traditional anti-tumor drugs.

[0007] In the first aspect, the present invention claims the application of a) or b) or c) or d) in any one of the following P1-P4:

[0008] a), plant JAZ protein.

[0009] b), a derived polypeptide of the plant JAZ protein.

[0010] c), a modified product of the plant JAZ protein described in a) or the derived polypeptide described in b); the modified product can be a product obtained by modifying the plant JAZ protein or the derived polypeptide by any one of the following modifications: glycosylation, phosphorylation, N-methylation, myristoylation, palmitoylation, biotinylation, fluorescent labeling, polyethylene glycol (PEG) modification, multiple antigen peptide (MAP), isoprenylation cyclization, cyclization and other modifications.

[0011] d), related biological materials of the plant JAZ protein described in a) or the derived polypeptide described in b); the related biological materials are any one of the following: 1) a nucleic acid molecule encoding the plant JAZ protein or the derived polypeptide; 2) an expression cassette, an expression vector or a recombinant microorganism containing the nucleic acid molecule.

[0012] P1, preparing a product for treating and / or preventing human or animal tumors.

[0013] P2, preparing a product for inhibiting the occurrence and / or development of human or animal tumors.

[0014] P3, preparing a product for inhibiting the proliferation of human or animal tumor cells.

[0015] P4, preparing a product for promoting the apoptosis of human or animal tumor cells.

[0016] In this application, the plant JAZ protein can interact with the MYC protein of humans or animals.

[0017] Furthermore, the MYC protein can be c-Myc protein, NMYC protein, and / or LMYC protein. Specifically, the plant JAZ protein can interact with the N-terminus of c-Myc protein through the NT domain.

[0018] In this application, the plant JAZ protein has all or part of the following functions: arresting the cell cycle of human or animal tumor cells at the G2-M phase; regulating c-Myc downstream genes in human or animal tumor cells; interacting with the cancer cell surface receptor uPAR (the plant JAZ protein can interact with the D3 domain of uPAR through the NT domain); interacting with the ligand uPA of the cancer cell surface receptor uPAR (interacting with the ATF domain of uPA); entering the nucleus of human or animal tumor cells.

[0019] In the present invention, the regulation of c-Myc downstream genes is specifically embodied as all or part of the following: downregulation of apoptosis-related genes Bcl2, BAX, HK2, FASN, MCM5 downstream of c-Myc, and upregulation of the P21 gene; downregulation of cell cycle-related genes CCNA2, CCNB1, CCND1, and upregulation of CCNE1.

[0020] Furthermore, the derivative polypeptide of the plant JAZ protein can be the NT domain, ZIM domain, or Jas domain of the plant JAZ protein, or a polypeptide containing the NT domain, ZIM domain, or Jas domain of the plant JAZ protein.

[0021] In a specific embodiment of the present invention, the plant JAZ protein is specifically a JAZ protein derived from cotton.

[0022] Furthermore, the JAZ protein derived from cotton can be any of the following:

[0023] (A1) A protein with an amino acid sequence shown in any one of SEQ ID No.1 to SEQ ID No.35;

[0024] (A2) A protein obtained by substituting and / or deleting and / or adding one or several amino acid residues to the amino acid sequence defined in (A1) and having the same function and derived from cotton;

[0025] (A3) A protein having more than 99%, more than 95%, more than 90%, more than 85%, or more than 80% identity with the amino acid sequence defined in any one of (A1)-(A2) and having the same function and derived from cotton;

[0026] (A4) A fusion protein obtained by connecting a protein tag to the N-terminus and / or C-terminus of the protein defined in any one of (A1)-(A3).

[0027] Among the above-mentioned proteins, the protein-tag refers to a polypeptide or protein that is fused and expressed with the target protein by using in vitro DNA recombination technology, so as to facilitate the expression, detection, tracing and / or purification of the target protein. The protein-tag can be a Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag, SUMO tag, etc.

[0028] Among the above-mentioned proteins, identity refers to the identity of amino acid sequences. The identity of amino acid sequences can be determined by using homology search sites on the Internet, such as the BLAST web page of the NCBI home page website. For example, in Advanced BLAST 2.1, by using blastp as the program, setting the Expect value to 10, setting all Filters to OFF, using BLOSUM62 as the Matrix, setting the Gap existence cost, Per residue gap cost and Lambda ratio to 11, 1 and 0.85 (default values) respectively, and performing a search to calculate the identity of a pair of amino acid sequences, and then the identity value (%) can be obtained.

[0029] Among the above-mentioned proteins, the homology of more than 95% can be identity of at least 96%, 97%, 98%. The homology of more than 90% can be identity of at least 91%, 92%, 93%, 94%. The homology of more than 85% can be identity of at least 86%, 87%, 88%, 89%. The homology of more than 80% can be identity of at least 81%, 82%, 83%, 84%.

[0030] Correspondingly, the nucleic acid molecule encoding the plant JAZ protein (i.e., the JAZ protein derived from cotton) can be any of the following:

[0031] (B1) A DNA molecule shown in any one of SEQ ID No. 36 to SEQ ID No. 70;

[0032] (B2) A DNA molecule that hybridizes with the DNA molecule defined in (B1) under stringent conditions and encodes the JAZ protein derived from cotton;

[0033] (B3) A DNA molecule that has a homology of more than 99%, more than 95%, more than 90%, more than 85% or more than 80% with the DNA sequence defined in any one of (B1)-(B2) and encodes the JAZ protein derived from cotton.

[0034] The DNA molecules shown in SEQ ID No. 36 to SEQ ID No. 70 encode the proteins shown in SEQ ID No. 1 to SEQ ID No. 35 in sequence.

[0035] In the above nucleic acid molecule, the stringent conditions may be as follows: Hybridization is carried out at 50°C in a mixed solution of 7% sodium dodecyl sulfate (SDS), 0.5 M Na3PO4 and 1 mM EDTA, and rinsing is carried out at 50°C in 2×SSC, 0.1% SDS; it may also be: Hybridization is carried out at 50°C in a mixed solution of 7% SDS, 0.5 M Na3PO4 and 1 mM EDTA, and rinsing is carried out at 50°C in 1×SSC, 0.1% SDS; it may also be: Hybridization is carried out at 50°C in a mixed solution of 7% SDS, 0.5 M Na3PO4 and 1 mM EDTA, and rinsing is carried out at 50°C in 0.5×SSC, 0.1% SDS; it may also be: Hybridization is carried out at 50°C in a mixed solution of 7% SDS, 0.5 M Na3PO4 and 1 mM EDTA, and rinsing is carried out at 50°C in 0.1×SSC, 0.1% SDS; it may also be: Hybridization is carried out at 50°C in a mixed solution of 7% SDS, 0.5 M Na3PO4 and 1 mM EDTA, and rinsing is carried out at 65°C in 0.1×SSC, 0.1% SDS; it may also be: Hybridization is carried out at 65°C in a solution of 6×SSC, 0.5% SDS, and then the membrane is washed once with 2×SSC, 0.1% SDS and once with 1×SSC, 0.1% SDS respectively.

[0036] In the above nucleic acid molecule, homology refers to the identity of nucleotide sequences. The identity of nucleotide sequences can be determined using homology search sites on the Internet, such as the BLAST web page on the NCBI home page website. For example, in Advanced BLAST 2.1, by using blastp as the program, setting the Expect value to 10, setting all Filters to OFF, using BLOSUM62 as the Matrix, setting the Gap existence cost, Per residue gap cost and Lambda ratio to 11, 1 and 0.85 (default values) respectively and performing a search to calculate the identity of a pair of nucleotide sequences, and then the identity value (%) can be obtained.

[0037] In the above nucleic acid molecule, the homology of more than 95% may be identity of at least 96%, 97%, 98%. The homology of more than 90% may be identity of at least 91%, 92%, 93%, 94%. The homology of more than 85% may be identity of at least 86%, 87%, 88%, 89%. The homology of more than 80% may be identity of at least 81%, 82%, 83%, 84%.

[0038] Furthermore, the human or animal tumors include but are not limited to the following: ovarian cancer, breast cancer, cervical cancer, lung cancer, etc.

[0039] Further, the human or animal tumor cells include, but are not limited to, the following: ovarian cancer cells, breast cancer cells, cervical cancer cells, lung cancer cells, etc.

[0040] In a specific embodiment of the present invention, the ovarian cancer cells are SKOV3 cells; the breast cancer cells are MCF7 cells; the cervical cancer cells are Hela cells; and the lung cancer cells are A549 cells.

[0041] In the above application, the product can be a drug.

[0042] Further, the drug is prepared by adding pharmaceutically acceptable excipients (such as excipients, diluents, etc.) on the basis of the above a), b), or c).

[0043] Further, the drug can also be a compound drug or an adjuvant drug.

[0044] Furthermore, the drug can be tablets, capsules, granules, syrups, etc. suitable for oral administration, or powder injections or solutions suitable for injection administration.

[0045] In a second aspect, the present invention claims the application of the above a), b), c), or d) in inhibiting the growth of plant callus.

[0046] Further, the plant callus is cotton callus.

[0047] In a specific embodiment of the present invention, the explant is a root segment or a hypocotyl.

[0048] In this application, the plant JAZ protein has all or part of the following functions: in the plant body, it interacts with NINJA protein, TPL protein, and HDA6 protein; in the presence of JA-Ile, it interacts with COI1 protein; and it can form a complex with MYC2 protein, NINJA protein, and TPL protein.

[0049] In a specific embodiment of the present invention, the plant is cotton.

[0050] In a third aspect, the present invention claims the application of JA synthesis inhibitors in inhibiting the growth of plant callus.

[0051] In a specific embodiment of the present invention, the JA synthesis inhibitor is copper reagent DICEA or ibuprofen.

[0052] Further, the plant callus is cotton callus.

[0053] In a specific embodiment of the present invention, the explant is a root segment or a hypocotyl.

[0054] Fourthly, the present invention claims to protect a method for inhibiting the growth of plant callus.

[0055] The method for inhibiting the growth of plant callus claimed by the present invention is to inhibit the growth of plant callus with the a), b), c), d) described in the first aspect above or the JA synthesis inhibitor described in the third aspect above.

[0056] In a specific embodiment of the present invention, the plant callus is cotton callus.

[0057] Furthermore, the method may include the following steps:

[0058] Experimental group: Overexpress the plant JAZ protein or the derivative polypeptide described in the first aspect above in the target plant to obtain transgenic positive plants; Take explants from the transgenic positive plants for tissue culture to obtain callus;

[0059] Control group: The difference from the experimental group is only that the explants taken during tissue culture are from the non-transgenic target plant;

[0060] The growth rate of the callus in the experimental group is slower than that in the control group.

[0061] Or,

[0062] Furthermore, the method may include the following steps:

[0063] Experimental group: Take explants from the target plant and place them on a callus induction medium supplemented with the JA synthesis inhibitor for tissue culture to obtain callus;

[0064] Control group: The difference from the experimental group is only that the callus induction medium used during tissue culture does not contain the JA synthesis inhibitor;

[0065] The growth rate of the callus in the experimental group is slower than that in the control group.

[0066] In a specific embodiment of the present invention, the explant is a root segment or a hypocotyl.

[0067] When the JA synthesis inhibitor is copper reagent DICEA, its final concentration in the callus induction medium can be 200 - 800 μM, such as 200 μM, 400 μM or 800 μM; When the JA synthesis inhibitor is ibuprofen, its final concentration in the callus induction medium is 20 - 80 μM, such as 20 μM, 40 μM or 80 μM.

[0068] Since cotton has a long tradition of being used as traditional Chinese medicine since ancient times (recorded in "Compendium of Materia Medica" etc.), and there are many JAZs in cotton. The plant-specific JAZ protein can block the activity of transcription factors such as MYC in plants, and MYC is highly conserved in evolution. Accordingly, the present invention proposes that cotton JAZ protein, JAZ homologous protein and their derived polypeptides, and modified products of the above proteins and polypeptides inhibit tumors in humans and animals by inhibiting human and animal proto-oncogenes MYCs (c-Myc, NMYC, LMYC), so as to achieve the purpose of preventing and treating tumors.

[0069] Experiments have proved that cotton JAZ protein, plant JAZ homologous protein and their derived polypeptides have significant inhibitory effects on human tumor cell lines such as human breast cancer MCF7, ovarian cancer SKOV3, cervical cancer Hela, and lung cancer A549; JAZ protein and its derived polypeptides have significant inhibitory effects on the occurrence and development of mouse in vivo tumors. Protein interaction experiments (yeast two-hybrid, immunoprecipitation, pull-down experiment, bimolecular fluorescence complementation, LUC experiment) confirmed that JAZ protein and its derived polypeptides have strong interactions with human proto-oncogenes MYCs (c-Myc, NMYC, LMYC). In summary, JAZ protein and its derived polypeptides can target various types of human and animal tumors and can be developed into targeted drugs for human and animal tumors. The present invention provides a new drug or adjuvant drug for the treatment of various human and animal tumors. Brief Description of the Drawings

[0070] Figure 1 Overexpression of GhJAZs inhibits the growth of cotton callus. A, "Zhongmiansuo 24" (WT) and callus overexpressing GhJAZ9 in "Zhongmiansuo 24" (left is the control WT, right is the overexpressing GhJAZ9 material), bar = 1 cm; B, enlarged view of the callus of wild-type "Zhongmiansuo 24", bar = 0.3 cm; C, enlarged view of the callus overexpressing GhJAZ9, bar = 0.3 cm; D, gene expression analysis of the GhJAZ9 gene in wild-type and overexpressing transgenic lines; E, callus weight statistics of wild-type and overexpressing transgenic lines; F, gene expression analysis of the GhJAZ1 gene in wild-type and overexpressing hairy roots of cotton; G, callus weight statistics of wild-type and overexpressing hairy roots of cotton; H, cell cycle gene expression analysis of callus in wild-type and overexpressing transgenic lines.

[0071] Figure 2Adding exogenous JA synthesis inhibitors can significantly inhibit the growth of cotton callus. A, Callus of "Zhongmian 24" grown on normal medium for 5 weeks; B, Callus of "Zhongmian 24" grown on medium containing DIECA for 5 weeks; C, Callus of Zhongmian 24 grown on medium supplemented with ibuprofen for 5 weeks; D, Statistical results of the weight of callus grown for 5 weeks (Ibu represents ibuprofen); E, Detection of JA content in callus (Ibu represents ibuprofen); F, Expression of cell cycle genes in callus on DIECA medium; G, Verification of the expression of cell cycle genes in callus on medium containing ibuprofen. Bar = 0.2 cm. In the figure, DIECA-1, DIECA-2, and DIECA-3 respectively represent three groups with addition amounts of 200 μM, 400 μM, and 800 μM in the callus induction medium in sequence; Ibu-1, Ibu-2, and Ibu3 respectively represent three groups with addition amounts of 20 μM, 40 μM, and 80 μM in the callus induction medium in sequence.

[0072] Figure 3 This is the distribution and phylogenetic tree of JAZ genes in various organisms. A, Evolutionary distribution of JAZ proteins in the biological world; B, Phylogenetic analysis of JAZ proteins in Arabidopsis and cotton.

[0073] Figure 4 This is the evolution and conserved domain analysis of MYC in the biological world. A, Evolutionary distribution of MYC proteins in the biological world; B and C, Conserved domain analysis of MYC proteins.

[0074] Figure 5 This is the interaction relationship between the main proteins and JAZ in the cotton JAZ protein transcriptional repression complex. A, Yeast two-hybrid interaction screening of cotton JAZ and MYC2 (AD represents the empty vector pGADT7 of yeast two-hybrid; BD represents the empty vector pGBKT7 of yeast two-hybrid, equivalent to the negative control); B, Luciferase experiment verification of the interaction between GhJAZ1 and GhMYC2; C, Luciferase experiment verification of the interaction between GhJAZ9 and GhMYC2; D, Verification of the interaction between mutants of GhJAZ1 and JAZ1 lacking NT, ZIM, and Jas respectively (marked as ΔNT, ΔZIM, and ΔJas in the figure represent the deletion of the corresponding domains) and GhMYC2, GhNINJA, GhTPL, GhHDA6, GhCOI (the upper side is without JA-Ile in the medium, and the lower side is with JA-Ile added in the medium). E, Luciferase experiment verification of the interaction between GhJAZ1 and GhNINJA; F, Luciferase experiment verification of the interaction between GhJAZ1 and GhCOI1; G, COIP experiment verification of the interaction between GhJAZ1 and GhTPL; H, COIP experiment verification of the interaction between GhJAZ1 and GhHDA6.

[0075] Figure 6It is the direct interaction between cotton JAZ and the proto-oncogene MYC. A. Similarities between plant "tumors" and animal tumors; B. Yeast two-hybrid interaction screening of cotton JAZ and MYC proto-oncogene (AD represents the empty vector pGADT7 of yeast two-hybrid; BD represents the empty vector pGBKT7 of yeast two-hybrid, equivalent to the negative control); C. Pull down verification of the interaction between GhJAZ1 and c-Myc; D. Co-IP verification of the interaction between GhJAZ1 and c-Myc; E. Co-IP verification of the interaction between GhJAZ1 and NMyc; F. LUC experiment verification of the interaction between GhJAZ1 and c-Myc; G. Bimolecular fluorescence complementation verification of the interaction between GhJAZ1 and c-Myc. Bar = 50 μm.

[0076] Figure 7 It is that the GhJAZ1 protein inhibits the proliferation of cancer cells. A. WB detection of the expression of GhJAZ1-His protein; B. The effect of GhJAZ1 on MCF7 cells, IC50 = 0.691 μM; C. The effect of GhJAZ1 on MCF10A cells, without inhibitory effect; D-G. Treatment of breast cancer cell line MCF7 with PBS; H-K. Treatment of breast cancer cell line MCF7 with GhJAZ1 protein (D and H: photographed under bright field; E and I: photographed under red fluorescence, and the red shows apoptotic cells; F and J: photographed under green fluorescence, and the green shows living cells; G and K: the result after the fusion of red fluorescence and green fluorescence); L. The effect of GhJAZ1 protein on A549 cells, IC50 = 0.697 μM; M. The effect of GhJAZ1 protein on Hela cells, IC50 = 0.553 μM; N. The effect of GhJAZ1 protein on SKOV3 cells, IC50 = 0.784 μM. Bar = 100 μm.

[0077] Figure 8 It is the influence of the GhJAZ1 protein on apoptosis and cell cycle of cancer cells and normal cells. A-C. The influence of the GhJAZ1 protein on apoptosis of MCF7 cells; D-F. The influence of the GhJAZ1 protein on apoptosis of MCF10A cells; G-I. The influence of the GhJAZ1 protein on the cell cycle of MCF7 cells; J-L. The influence of the GhJAZ1 protein on the cell cycle of MCF10A cells. Among them, A, D, G, J are the control groups (added with PBS), B, E, H, K are the GhJAZ1 treatment groups, and C, F, I, L are the statistical data.

[0078] Figure 9Regulation of c-Myc downstream genes by GhJAZ1. A, Heat map analysis of differentially expressed genes in MCF7 cells treated with PBS and GhJAZ1 protein; B, Detection of the expression levels of differentially expressed genes in MCF7 cells treated with PBS and GhJAZ1 protein; C, Detection of the expression levels of c-Myc downstream genes in MCF7 cells treated with PBS and GhJAZ1 protein; D, Detection of the expression levels of c-Myc downstream genes in MCF10A cells treated with PBS and GhJAZ1 protein.

[0079] Figure 10 Direct interaction between GhJAZ1 and cancer cell surface receptors. A, Yeast two-hybrid screening of the interaction between GhJAZ1 and cancer cell surface receptors (AD represents the empty vector pGADT7 of yeast two-hybrid; BD represents the empty vector pGBKT7 of yeast two-hybrid, equivalent to the negative control); B, COIP interaction verification of the interaction between GhJAZ1 and the cancer cell surface receptor uPAR; C and D, Interaction analysis of uPAR and GhJAZ1 with the ATF domain of uPA respectively.

[0080] Figure 11 Experimental verification of the entry of GhJAZ1 protein into cells and the nucleus. A, WB detection of GhJAZ1+RFP-His protein. Lane M: Protein blot marker; Lane NC: Cell lysate without induction; Lane 1: Cell lysate induced at 15 °C for 16 hours; Lane 2: Cell lysate induced at 37 °C for 4 hours; Lane 3: Supernatant of cell lysate induced at 15 °C for 16 hours; Lane 4: Supernatant of cell lysate induced at 37 °C for 4 hours; Lane 5: Fragment of cell lysate induced at 15 °C for 16 hours; Lane 6: Fragment of cell lysate induced at 37 °C for 4 hours. The primary antibody for immunoblotting is anti-histidine antibody (GenScript, Cat. No. A00186); B-E, Treatment of breast cancer cell MCF7 with PBS; F-I, Treatment of breast cancer cell MCF7 with IRP-803 (uPAR inhibitor); J-M, Treatment of breast cancer cell MCF7 with GhJAZ1 protein (B, F and J: Photographed under bright field; C, G and K: Photographed under blue fluorescence, and the blue shows the nucleus; D, H and L: Photographed under red fluorescence, H shows the red fluorescence of IRP-803 outside the nucleus, and L shows the red fluorescence of GhJAZ1+GFP, which is dot-like distributed; E, I and M: Results of the fusion of blue fluorescence and red fluorescence).

[0081] Figure 12 Results of the anti-living tumor experiment of GhJAZ1 protein. A, Photographs of living tumors in the drug injection group and the control group; B, Statistical analysis of the volumes of living tumors in the drug injection group and the control group; C, Photographs of living tumors in the drug injection group and the control group; D, Statistical analysis of the weights of living tumors in the drug injection group and the control group.

[0082] Figure 13 Results of experiments on the interaction domains of GhJAZ1 with uPAR and c-Myc. A. Yeast two-hybrid interaction analysis of GhJAZ1 and yeast vectors lacking the NT, ZIM, and JAS domains with the ATF domains of uPAR and uPA, respectively; B. Interaction analysis of uPAR and yeast vectors lacking the D1, D2, and D3 domains with GhJAZ1, respectively; C. Interaction analysis of GhJAZ1 and yeast vectors lacking the NT, ZIM, and JAS domains with full-length c-Myc and NMYC, respectively; D. Interaction analysis of c-Myc and c-Myc truncated 150 and 215 amino acids, respectively, with GhJAZ1; E. Verification of the interaction of c-Myc and c-Myc truncated 150 and 215 amino acids, respectively, with the LUC of GhJAZ1. DETAILED DESCRIPTION

[0083] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0084] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials and reagents used in the following examples, unless otherwise specified, were all commercially available.

[0085] Example 1: Analysis of the effect of overexpression of cotton JAZ on callus growth

[0086] 1. Creation of transgenic materials overexpressing GhJAZ1

[0087] Based on the full-length and domain conservation analysis of the Arabidopsis JAZ1 amino acid sequence in the cotton database, the cotton JAZ1 with the highest homology to AtJAZ1 was identified, with the gene number Gh_A05G0260, and the sequence is shown in SEQ ID No. 4. The full-length CDS of GhJAZ1 (SEQ ID No. 39) was ligated between the XbaI and PacI restriction sites of the modified pCAMBIA2300 (the original pCAMBIA2300 vector is a product of Youbao Biotechnology, catalog number: VT1383, and the specific modification is: replacing the MCS region of the original pCAMBIA2300 vector with the 2X35S-YFP sequence, and the 2X35S-YFP sequence is shown in SEQ ID No. 89). The recombinant positive plasmid verified by sequencing was transformed into Agrobacterium tumefaciens K599. Using cotton "Zhongmiansuo 24" as the material, transgenic hairy root cotton materials were created by the genetic transformation technique mediated by Agrobacterium rhizogenes. The steps were as follows: culturing sterile seedlings, preparing cotyledons, infecting cotton cotyledons with the Agrobacterium solution, and then growing roots. The obtained resistant roots were identified at the RNA level.

[0088] RNA level identification: RNA was extracted using an RNA extraction kit (RNAprep Pure Polysaccharide and Polyphenol Plant Total RNA Extraction Kit (DP441), manufacturer: Tiangen Biochemical Technology (Beijing) Co., Ltd.), and then cDNA was obtained using a two-step reverse transcription kit (a three-generation premix for removing genomic DNA and reverse transcription, catalog number MR05201). Using cDNA as a template, it was added to a 96-well plate, and Green I chimeric dye (Antibody Dye-based Quantitative PCR Premix, catalog number MQ10101) and gene primers were added for three-step qPCR amplification. Finally, the relative gene expression level was calculated based on the Ct value.

[0089] Using Actin as an internal reference, the detection primers are:

[0090] Q-Actin-F: 5’-GACCCAGATCATGTTTGAGACCT-3’;

[0091] Q-Actin-R: 5’-CAGTGTGGCTGACACCATCAC-3’.

[0092] The primers used to detect GhJAZ1 are:

[0093] Q-GhJAZ1-F: 5’-GATGATAGTCCCAACAAGTTGGAGCC-3’;

[0094] Q-GhJAZ1-R: 5’-TCGCTTCTCTAGGAACCGATGCA-3’.

[0095] The results showed that OEGhJAZ1-1 and OEGhJAZ1-8 were significantly increased compared with the control, which was 70-300 times that of the control ( Figure 1 (F) OEGhJAZ1-1 and OEGhJAZ1-8 are two positive lines randomly selected from transgenic progeny.

[0096] 2. Creation of transgenic materials overexpressing GhJAZ9

[0097] Based on the full-length and domain conservation analysis of the Arabidopsis JAZ9 amino acid sequence in the cotton database, the cotton JAZ9 with the highest homology to AtJAZ9 was identified, and the sequence is shown in SEQ ID No. 2. The full-length CDS of GhJAZ9 (SEQ ID No. 37) was ligated into the vector 8 / GW / TOPO (Invitrogen, Catalog No.: K252020) between EcoRI and EcoRI, through LR reaction (ThermoFisher, Gateway LR Clonase Enzyme mix, Catalog No. 11791019) connected to the pEarleyGate 303 vector (Ubao Bio, Catalog No.: VT9027), the recombinant positive plasmid verified by sequencing was transformed into Agrobacterium LBA4404, and the cotton "Zhongmian Institute 24" was used as the material to create transgenic materials using Agrobacterium-mediated genetic transformation technology. The steps are to culture sterile seedlings, prepare explants, infect cotton explants with Agrobacterium liquid, and then dedifferentiate to form resistant callus tissue, regenerate seedlings, and finally obtain resistant plants. The obtained resistant plants were identified at the RNA level. The RNA identification method is the same as above, and the primers used to detect GhJAZ9 are as follows:

[0098] Q-CO39-F1: 5'-CTTCATGCCTATATCAACAATGGATGCT-3';

[0099] Q-CO39-R1: 5'-CAAGGGTGGGAAGAACTGAATGTG-3'.

[0100] The results showed that the expression levels of OEGhJAZ9-2, OEGhJAZ9-3 and OEGhJAZ9-6 were significantly increased compared with the control group ( Figure 1 Middle D) Among them, OEGhJAZ9-2, OEGhJAZ9-3 and OEGhJAZ9-6 are three positive strains randomly selected from the transgenic progeny.

[0101] 3. Effect of cotton JAZ overexpression on callus growth

[0102] Cut the overexpressed positive roots and negative roots of GhJAZ1 (i.e., the roots of CCRI 24 transformed with the empty pCAMBIA2300 vector) identified in Step 1 into explants of the same length, and place them in the same culture dish for simultaneous cultivation. After one month, observe the growth of the callus, take photos with a camera and a microscope, measure the weight of the callus, and collect samples for fluorescence quantitative PCR to detect the expression of cell cycle-related genes.

[0103] The steps of RNA extraction, reverse transcription, and fluorescence quantitative PCR are the same as above, and the primer sequences are as follows:

[0104] Primers for detecting GhCYCA1:

[0105] Q-GhCYCA1-1-F: 5’-GTACAACCCGGTTCAGTTCCG-3’;

[0106] Q-GhCYCA1-1-R: 5’-GAATCAACAGCCGTAACATCATG-3’.

[0107] Primers for detecting GhCYCA2:

[0108] Q-GhCYCA2-4-F: 5’-GCACTTAATGGTCGAATCACACGT-3’;

[0109] Q-GhCYCA2-4-R: 5’-ATTATTGCAGCAAACATTTGTGACAT-3’. [[ID=2,3]]

[0110] Primers for detecting GhCYCB:

[0111] Q-GhCYCB-F: 5’-GGCTCAGATTCAGACTCATCGTC-3’;

[0112] Q-GhCYCB-R: 5’-CAGCAGGAGCAGCAACACTTC-3’.

[0113] Primers for detecting GhCYCD1-1:

[0114] Q-GhCYCD1-1-F: 5’-CTTCACGGACTTACTCTGCGC-3’;

[0115] Q-GhCYCD1-1-R: 5’-CACTGAAACCGAGCAAGGTAATC-3’.

[0116] Primers for detecting GhCYCD3-1:

[0117] Q-GhCYCD3-1-F: 5’-ATGGCAATACAGCAATATGAACAGC-3’;

[0118] Q-GhCYCD3-1-R: 5’-CCTGCTCTAACAACAACAGTGGG-3’.

[0119] Primers for detecting GhCYCD6-1:

[0120] Q-GhCYCD6-1-F: 5’-ACCCATTAACAAACTTCGATGACTT-3’;

[0121] Q-GhCYCD6-1-R: 5’-TCGAGATAATTGACAGCAAGATACG-3’.

[0122] Sterile seedlings were separately cultured from the seeds of the homozygous GhJAZ9 overexpression transgenic lines identified in step 2 and the seeds of "Zhongmiansuo 24". After 7 days, the hypocotyl segments were taken and cultured on the callus induction medium (without hormones) to induce callus. The explants of the GhJAZ9 overexpression lines and the explants of "Zhongmiansuo 24" were cut to the same length and placed in the same culture dish for simultaneous culture. After one month, the growth of the callus was observed, and photos were taken with a camera and a microscope, and the weight of the callus was counted. At the same time, the GhJAZ9 overexpressing callus was sampled for fluorescence quantitative PCR to detect the expression of cell cycle-related genes (the method steps and primers are the same as above).

[0123] The results showed that the callus growth of JAZ overexpressing cotton was slow, and the expression of cell cycle regulatory genes in callus cells was severely affected ( Figure 1 ), indicating that overexpression of cotton JAZ can inhibit the growth of cotton callus.

[0124] Example 2. Analysis of the effect of JA synthesis inhibitor on callus growth

[0125] The JA signaling pathway may also be the key signaling pathway regulating the formation of cotton callus. Reported JA synthesis inhibitors internationally include copper reagent DICEA and ibuprofen, etc. Sterile seedlings were cultured from the seeds of "Zhongmiansuo 24". After 7 days, the hypocotyl cuttings were taken and cultured for callus on callus induction media (control (not added); added DIECA (also known as copper reagent, Sangon Biotech, catalog number A600846), final concentrations set at 200 μM, 400 μM, 800 μM; added ibuprofen (also known as Ibuprofen, catalog number: HY-78131), final concentrations set at 20 μM, 40 μM, 80 μM). The explant cuttings of "Zhongmiansuo 24" had the same length and were cultured under the same conditions. After one month, the growth of the callus was observed, and photos were taken with a camera and microscope, the weight of the callus was counted, the JA content was measured, and samples were taken for fluorescence quantitative PCR to detect the expression of cell cycle-related genes (the method steps and primers were the same as in Example 1).

[0126] Method for measuring JA content:

[0127] 1) Method for extracting jasmonic acid: Weigh approximately 0.1 g of the sample, add 1 mL of pre-cooled Reagent 1 (80% acetonitrile), extract overnight at 4°C, centrifuge at 8000 g for 10 min. The residue was extracted with 0.5 mL of Reagent 1 (80% acetonitrile) for 2 hours. After centrifugation, the supernatant was taken out, and the two supernatants were combined. Nitrogen was blown dry at 40°C, 0.5 mL of water was added for reconstitution, 0.5 mL of Reagent 2 (petroleum ether) was added for extraction and decolorization at 60°C - 90°C for 3 times, and the upper ether phase was discarded. Reagent 3 (saturated citric acid aqueous solution) was added to adjust the pH to 2.5 - 3.0. After mixing, it was extracted three times with an equal volume of Reagent 4 (ethyl acetate), and the organic phases were combined and nitrogen was blown to dryness. 0.2 mL of Reagent 5 (ether:methanol = 9:1, volume ratio) was added for reconstitution, 20 μL of Reagent 6 (2 mmol / L trimethylsilyldiazomethane (dissolved in n-hexane)) was added, and after mixing, it was placed at room temperature (25°C) for 30 min. 20 μL of Reagent 7 (2 mol / L acetic acid n-hexane solution) was added, and after mixing, it was placed at room temperature (25°C) for 30 min. Nitrogen was blown dry in an ice-water bath. 0.5 mL of methanol was added, vortexed and shaken for dissolution, and filtered through a syringe filter for measurement.

[0128] 2) HPLC liquid phase conditions: Chromatograph: RIGOL L3000 high performance liquid chromatograph, ultraviolet wavelength 210 nm.

[0129] Chromatographic column: RIGOL C18 reversed-phase chromatographic column (250 mm * 4.6 mm, 5 μm)

[0130] Column temperature: 35°C, flow rate: 0.8 mL / min, injection volume: 10 μL. Preparation of the mobile phase: Mobile phase A: acetonitrile, B: 0.1% phosphoric acid aqueous solution, isocratic elution, (60% of A + 40% of B, % represents volume percentage).

[0131] The results are as follows Figure 2 shown: Both DIECA and ibuprofen can significantly inhibit the growth of cotton callus, the JA content also decreases significantly, and the cell cycle-related genes of the callus also change significantly, indicating that JA synthesis inhibitors can all inhibit the growth of cotton callus.

[0132] Example 3, Evolutionary analysis and functional study of JAZ genes in plants

[0133] 1. Distribution and evolutionary analysis of JAZ and MYC genes in various organisms

[0134] The MEGA7 software was used to perform gene homology analysis on JAZ proteins in the biological world and draw phylogenetic trees. The analysis results are as follows Figure 3 shown in A: JAZ only exists in autotrophic organisms (such as 9 in Physcomitrella patens of Bryophyta, 1 in Marchantia polymorpha of liverworts, 2 in Cycas micholitzii of gymnosperms, 7 in Spirodela polyrhiza of monocotyledons, and 35 in Gossypium hirsutum of dicotyledons), while it does not exist in mixotrophic organisms (such as Chlamydomonas reinhardtii) and heterotrophic organisms (such as viruses, bacteria, fungi, lower animals, higher animals, and humans). A total of 35 JAZ members were identified in Gossypium hirsutum (the amino acid sequences are shown as SEQ ID No.1 to SEQ ID No.35, and the corresponding coding genes are shown as SEQ ID No.36 to SEQ ID No.70 in sequence). According to the evolutionary analysis, they can be mainly divided into 6 major categories represented by GhJAZ1, GhJAZ9, GhJAZ7, GhJAZ10, GhJAZ11, and GhJAZ31 ( Figure 3 shown in B).

[0135] MYC belongs to the bHLH transcription factor family that exists ubiquitously in the biological world. From the perspective of evolutionary relationships, MYC exists in viruses, prokaryotes, and eukaryotes ( Figure 4 shown in A). The most studied in plants is MYC2, which acts as a regulatory hub within the JA signaling pathway. It contains a conserved bHLH domain in its carboxyl domain, which can form homodimers or heterodimers with other proteins and bind to the G-box (5'-CACGTG-3') in the promoter of target genes. MYC2 contains a transcriptional activation domain (TAD) at its amino terminus, which recruits the mediator complex required for transcriptional initiation through its TAD, and this complex specifically interacts with the plant mediator complex MED25. From the results of amino acid homology alignment, MYC is relatively conserved in the TAD and bHLH domains ( Figure 4 shown in B and C).

[0136] 2. Study on the proteins and protein complexes that interact with JAZ genes in cotton

[0137] Studies on Arabidopsis thaliana have shown that the inhibition of JA signaling by JAZ proteins is mainly achieved by inhibiting the transcriptional factor function of MYC. We verified this result through experiments in cotton. Based on the phylogenetic tree of JAZ in Arabidopsis thaliana and cotton, we screened six cotton JAZs (GhJAZ1, GhJAZ7, GhJAZ9, GhJAZ10, GhJAZ11, GhJAZ31) for yeast two-hybrid analysis with GhMYC2, GhNINJA, GhTPL, GhHDA6, and GhCOI1. The amino acid sequence of GhJAZ1 is shown in SEQ ID No. 4, and the corresponding coding gene sequence is shown in SEQ ID No. 39; the amino acid sequence of GhJAZ7 is shown in SEQ ID No. 7, and the corresponding coding gene sequence is shown in SEQ ID No. 42; the amino acid sequence of GhJAZ9 is shown in SEQ ID No. 2, and the corresponding coding gene sequence is shown in SEQ ID No. 37; the amino acid sequence of GhJAZ10 is shown in SEQ ID No. 3, and the corresponding coding gene sequence is shown in SEQ ID No. 38; the amino acid sequence of GhJAZ11 is shown in SEQ ID No. 5, and the corresponding coding gene sequence is shown in SEQ ID No. 40; the amino acid sequence of GhJAZ31 is shown in SEQ ID No. 9, and the corresponding coding gene sequence is shown in SEQ ID No. 44. The gene ID of GhMYC2 is Gh_A12G1893 (https: / / cottonfgd.org / ); the gene ID of GhNINJA is Gh_D05G0252 (https: / / cottonfgd.org / ); the gene ID of GhTPL is Gh_D05G1161 (https: / / cottonfgd.org / ); the gene ID of GhHDA6 is Gh_D03G0986 (https: / / cottonfgd.org / ); the gene ID of GhCOI1 is Gh_A05G2749 (https: / / cottonfgd.org / ).

[0138] Yeast two-hybrid assay method: Using BamH1 and EcoR1 restriction endonucleases to construct PGBKT7::GhJAZ1 (the DNA fragment shown in SEQ ID No. 39 is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid), PGBKT7::GhJAZ9 (the DNA fragment shown in SEQ ID No. 37 is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid), PGBKT7::GhJAZ7 (the DNA fragment shown in SEQ ID No. 42 is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid), PGBKT7::GhJAZ10 (the DNA fragment shown in SEQ ID No. 38 is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid), PGBKT7::GhJAZ11 (the DNA fragment shown in SEQ ID No. 40 is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid), PGBKT7::GhJAZ31 (the DNA fragment shown in SEQ ID No. 44 is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid), PGBKT7::GhJAZ1 ΔNT (GhJAZ1 shown in SEQ ID No. 74 ΔNT gene sequence is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid, GhJAZ1 ΔNT protein sequence is as shown in SEQ ID No. 73), PGBKT7::GhJAZ1 ΔZIM (GhJAZ1 shown in SEQ ID No. 76 ΔZIM gene sequence is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid, GhJAZ1 ΔZIM protein sequence is as shown in SEQ ID No. 75), PGBKT7::GhJAZ1 ΔJas (GhJAZ1 shown in SEQ ID No. 78 ΔJas gene sequence is inserted between BamH1 and EcoR1 of the pGBKT7 plasmid, GhJAZ1 ΔJasCotton JAZ yeast plasmids such as the protein sequence shown in SEQ ID No. 77), etc., and PGADT7::GhMYC2 (the DNA fragment of gene number Gh_A12G1893 (https: / / cottonfgd.org / ) was inserted between BamH1 and EcoR1 of the pGADT7 plasmid), PGADT7::GhNINJA (the DNA fragment of gene number Gh_D05G0252 (https: / / cottonfgd.org / ) was inserted between BamH1 and EcoR1 of the pGADT7 plasmid), PGADT7::GHTPL (the DNA fragment of gene number Gh_D05G1161 (https: / / cottonfgd.org / ) was inserted between BamH1 and EcoR1 of the pGADT7 plasmid), PGADT7::GhHDA6 (the DNA fragment of gene number Gh_D03G0986 (https: / / cottonfgd.org / ) was inserted between BamH1 and EcoR1 of the pGADT7 plasmid), PGADT7::GhCOI1 (the DNA fragment of gene number Gh_A05G2749 (https: / / cottonfgd.org / ) was inserted between BamH1 and EcoR1 of the pGADT7 plasmid), etc. After the vectors were verified to be correct by sequencing, they were co-transformed (one was pGBKT7 + gene plasmid, and the other was pGADT7 + gene plasmid) into the yeast competent Y2HGold (Shanghai Weidi Co., Ltd., product number: YC1002). Carrier DNA (95°C, 5 min), PEG / LiAC were added and mixed evenly. Incubate in a water bath at 30°C for 30 min (invert 6 - 8 times to mix evenly at 15 min), incubate in a water bath at 42°C for 15 min (invert 6 - 8 times to mix evenly at 7.5 min), centrifuge at 5000 rpm for 40 S, discard the supernatant, resuspend with ddH2O, centrifuge at 5000 rpm for 30 S, discard the supernatant, resuspend with ddH2O, and spread on the double-deficient medium (SD / -Leu / -Trp). Culture at 28°C for 1 - 2 days. After colonies appeared, pick the colonies and transfer them to the quadruple-deficient medium (SD-Ade / -His / -Leu / -Trp, supplemented with Aba, X-α-gal). Observe the interaction situation after culturing at 28°C for 3 - 4 days.

[0139] Luciferase assay method: Subclone GhJAZ1 (the corresponding coding gene sequence is shown in SEQ ID No. 39) between the KpnI and SalI restriction sites of the pCAMBIA-NLuc vector (Chen, H., et al. "Firefly Luciferase Complementation Imaging Assay for Protein-Protein Interactions in Plants." Plant Physiology 146.2 (2008): 368-376.). Subclone transcription factors such as GhMYC2 (gene number: Gh_A12G1893 (https: / / cottonfgd.org / )), GhNINJA (gene number: Gh_D05G0252 (https: / / cottonfgd.org / )), and GhCOI1 (gene number: Gh_A05G2749 (https: / / cottonfgd.org / )) into the KpnI and SalI restriction sites of pCAMBIA-NLuc (Chen, H., et al. "Firefly Luciferase Complementation Imaging Assay for Protein-Protein Interactions in Plants." Plant Physiology 146.2 (2008): 368-376.) respectively. Then transform the constructed plasmids into Agrobacterium tumefaciens GV3101 (pSoup-p19) (Shanghai Weidi Biotechnology Co., Ltd., product number: AC1003). Incubate on ice for 5 min, in liquid nitrogen for 5 min, in a 37°C water bath for 5 min, on ice for 5 min. Add LB liquid medium and culture at 28°C, 220 rpm for 2-3 h, then spread on an LB solid plate containing the corresponding antibiotics (kanamycin + rifampicin) and culture at 28°C for 2-3 days. After colonies appear, pick positive monoclonal colonies for culture. Then mix the corresponding NLUC and CLUC vectors and inject them into the leaves of tobacco (1 day in the dark / 1 day in the light). Before taking pictures, apply luciferase substrate to the infected leaves, place them in a dark room for at least 6 min to quench fluorescence, and then observe under the luciferase-5min condition by clicking on the IndigoBasic software in the Nightowl-II LB983 instrument.

[0140] COIP experimental method steps: Add non-denaturing lysis buffer to a 293T cell (Procell, catalog number: CL-0005) culture plate and completely lyse the cells at 4°C. Centrifuge the lysate at 12,000 rpm for 10 minutes and collect the supernatant. Add the corresponding antibody to the obtained non-denaturing protein solution and incubate it overnight at 4°C. Wash Pierce TM Protein A / G Agarose Beads (Thermo Fisher Scientific, Inc.) with lysis buffer, add the pretreated beads to the cell lysate, and incubate at room temperature for 2 hours. Centrifuge the mixture at 2500 rpm for 5 minutes at 4°C, remove the supernatant, and wash the agarose beads 5 times with 1 ml of lysis buffer. Add an appropriate amount of protein loading buffer to the pellet and incubate the sample in a 100°C water bath for 10 minutes. Detect the target band by Western Blot.

[0141] Yeast two-hybrid results showed that GhJAZ1 interacted strongly with GhMYC2, and GhJAZ9 interacted weakly with GhMYC2 ( Figure 5 in A). Luciferase assay results showed that when Agrobacterium mixtures of GhJAZ1 and GhMYC2 were co-injected into tobacco, strong fluorescence could be observed after applying the luciferase substrate, while the control had no fluorescence ( Figure 5 in B); when Agrobacterium mixtures of GhJAZ9 and GhMYC2 were co-injected into tobacco, weak fluorescence could be observed after applying the luciferase substrate, while the control had no fluorescence ( Figure 5 in C); which further demonstrated that there was a strong interaction between GhJAZ1 and GhMYC2, and a weak interaction between GhJAZ9 and GhMYC2.

[0142] Yeast two-hybrid was used to explore the interaction relationship between GhJAZ1 and the main proteins in the JA signaling pathway transcriptional repression complex. The results showed that GhJAZ1 interacted with GhNINJA, GhTPL, and GhHDA6 ( Figure 5 in D), and in the presence of JA-Ile, GhJAZ1 interacted with GhCOI1 ( Figure 5 in D); Luciferase assay results showed that when Agrobacterium mixtures of GhJAZ1 and GhNINJA were co-injected into tobacco, relatively bright fluorescence could be observed after applying the luciferase substrate, while the control had no fluorescence ( Figure 5 in E); when Agrobacterium mixtures of GhJAZ1 and GhCOI1 were co-injected into tobacco, weak fluorescence could be observed after applying the luciferase substrate, while the control had no fluorescence ( Figure 5 in F), proving that GhJAZ1 interacted with GhNINJA and GhCOI1. COIP results showed that GhJAZ1 interacted with GhTPL and GhHDA6 ( Figure 5In G and H). The above results indicate that in cotton, JAZ1 forms a transcriptional repression complex with MYC2, NINJA, TPL, etc., thereby inhibiting the transcriptional activity of MYC2.

[0143] Exploring the interacting functional domains is crucial for developing functional polypeptides. The interaction relationships between GhJAZ1 lacking functional domains and GhMYC2, GhNINJA, GhTPL, GhHDA6, and GhCOI1 were studied by yeast two-hybrid. The results showed that the interaction between GhJAZ1 lacking the NT and ZIM domains and GhMYC2 was significantly weakened; the interaction between GhJAZ1 lacking the ZIM domain and GhNINJA was significantly weakened; the interaction between GhJAZ1 lacking the NT and Jas domains and GhTPL was significantly weakened; the interaction between GhJAZ1 lacking the NT domain and GhHDA6 was significantly weakened; the interactions between GhJAZ1 lacking the NT, ZIM, and Jas domains and GhCOI1 were all significantly weakened, proving that the NT of GhJAZ1 is mainly involved in the interactions with GhMYC2, GhTPL, GhHDA6, and GhCOI1, the ZIM domain is involved in the interactions with GhMYC2, GhNINJA, and GhCOI1, and the Jas domain is involved in the interactions with GhTPL and GhCOI1 ( Figure 5 In D).

[0144] Example 4. Analysis of the anti-tumor effect of cotton JAZ protein

[0145] 1. Analysis of the similarities and differences between plant "tumors" and animal tumors

[0146] There are many similarities between plant callus (plant tumor) and human tumors, such as similar morphology; induced by endogenous and exogenous stimuli; having persistent regeneration and self-replication abilities; relatively fast cell division, rapid self-proliferation, and unstable cell growth; both being regulated by the transcription factor MYC, etc. The growth of "plant tumors" is strictly regulated by the JA signal and cannot develop malignantly. Different from the controllable "plant tumors", human tumors often deteriorate and develop into cancer due to uncontrollability. Plant callus cells are totipotent and can differentiate into various other tissue cells; while the cells differentiated from cancer cells are still cancer cells, and they also have the characteristics of loss of contact inhibition, weakened adhesion between cancer cells, and decreased adhesion to the wall ( Figure 6 In A).

[0147] 2. Screening and verification of the interaction between cotton JAZ protein and human MYC protein

[0148] The coding genes of 6 JAZ representatives of cotton (GhJAZ1, GhJAZ7, GhJAZ9, GhJAZ10, GhJAZ11, GhJAZ31, for the specific sequences, see Example 3) were respectively constructed between EcoRI and BamHI of the pGBKT7 vector (Youbao Biotech, product number: VT1638). Meanwhile, the MYC oncogenes (c-Myc (NCBI accession number: NM_002467.6), NMYC (NCBI accession number: NP_005369.2), and LMYC (NCBI accession number: NP_001028254.2)) were respectively constructed between EcoRI and BamHI of the pGADT7 vector (Youbao Biotech, VT1639) for yeast two-hybrid interaction analysis (the method is the same as that in Example 3). The results showed that GhJAZ1 interacted with both c-Myc and NMYC ( Figure 6 in B).

[0149] Meanwhile, the interaction was verified by GST Pull down experiment. First, construct pET-30a-GhJAZ1 (insert the DNA sequence shown in SEQ ID No. 39 between NdeI and HindIII of the pET-30a(+) vector (Youbao Biotech, product number: VT1212)) and pGEX-4T-1-c-Myc vector (insert the CDS sequence of c-Myc (NCBI accession number: NM_002467.6) between BamHI and EcoRI of the pGEX-4T-1 vector (Youbao Biotech, product number: VT1253)), transform Escherichia coli BL21, incubate on ice for 30 min, heat shock at 42 °C for 60 s, incubate on ice for 2 min, add LB medium, culture at 37 °C and 200 rpm for 1 h, spread on LB plates with corresponding antibiotics, and culture at 37 °C. After colonies appeared, pick monoclonal colonies for shaking culture, and then expand the culture. When the medium began to turn slightly white, add IPTG at different concentrations for induction and set different temperatures for overnight culture. Then centrifuge at 5000 rpm and 4 °C for 10 min, resuspend with an equal volume of PBS (add PMSF and DTT). Ultrasonically disrupt it, disrupt for 5 s and incubate on ice for 5 s until the resuspended solution becomes significantly clear. Centrifuge at 13,000 rpm and 4 °C for 10 min to separate the supernatant and inclusion bodies. Transfer the supernatant to a protein purification column, incubate, wash, and elute, and detect and collect the protein drop by drop using a protein prestained solution. After obtaining the GST-c-Myc fusion protein and His-GhJAZ1 fusion protein through the above expression and purification, wash the GST beads, add 2 μg of the GST-c-Myc fusion protein for incubation for 2 h; then wash again, add 1 μg of the His-GhJAZ1 fusion protein, and aspirate a part as Input and incubate for 1 h; finally, wash again and then add loading buffer to run WB to detect the results. The results also proved that GhJAZ1 interacted with c-Myc (Figure 6 in C).

[0150] Using the co-immunoprecipitation method (the experimental method was the same as in Example 3). The COIP results showed that GhJAZ1 interacted with c-Myc and NMYC in HEK293 cells ( Figure 6 in D and E).

[0151] Using the LUC experimental method, c-Myc (NCBI accession number: NM_002467.6) was subcloned between the KpnI and SalI restriction sites of the pCAMBIA-NLuc vector (Chen, H., et al. "Firefly Luciferase Complementation Imaging Assay for Protein-Protein Interactions in Plants." Plant Physiology 146.2 (2008): 368-376.), and GhJAZ1 (SEQ ID No. 39) was subcloned between the KpnI and SalI restriction sites of the pCAMBIA-CLuc vector (Chen, H., et al. "Firefly Luciferase Complementation Imaging Assay for Protein-Protein Interactions in Plants." Plant Physiology 146.2 (2008): 368-376.). The constructed recombinant plasmid verified positive by sequencing was transformed into Agrobacterium tumefaciens GV3101 (pSoup-p19) (Shanghai Weidi Biotechnology Co., Ltd., product number: AC1003). After co-transforming tobacco leaves, 2-3 days after injection, before taking pictures, the infected leaves smeared with luciferin substrate were placed in the darkroom of the machine for at least 6 min to quench the fluorescence. Subsequently, on the Nightowl-II LB983 instrument, click on the IndigoBasic software and enter the luciferase-5min condition for observation. After co-infection and cultivation, if there is an interaction, the LUC luciferase will be expressed, and luminescence can be detected after smearing the substrate. As Figure 6 shown in F, GhJAZ1 interacted with c-Myc.

[0152] Through bimolecular fluorescence complementation experiments, GhJAZ1 (SEQ ID No. 39) was subcloned between the BamHI and XbaI restriction sites of the pUC-SPYNE vector (Song, Yun, et al. "BIN2 negatively regulates plant defense against Verticillium dahliae in Arabidopsis and cotton." Plant Biotechnology Journal (2021)) to construct pUC-SPYNE-GhJAZ1. c-Myc (NCBI accession number: NM_002467.6) was subcloned between the BamHI and XbaI restriction sites of the pUC-SPYCE vector (Song, Yun, et al. "BIN2 negatively regulates plant defense against Verticillium dahliae in Arabidopsis and cotton." Plant Biotechnology Journal (2021)) to construct pUC-SPYCE-c-Myc. pUC-SPYNE-GhJAZ1 and pUC-SPYCE-c-Myc were co-transformed into Agrobacterium tumefaciens GV3101 for transient infiltration of tobacco leaf tissues, and YFP signals were observed under a laser confocal microscope. As shown in G of Figure 6 , there was an interaction between GhJAZ1 and c-Myc in the tobacco cell nucleus.

[0153] 3. Test on the inhibitory effect of GhJAZ1 protein on tumor cells

[0154] CCK8 cell viability assay: A cell suspension with a concentration of 5×10 4 cells / ml (100 μL / well) was seeded in a 96-well plate. The culture plate was pre-incubated in an incubator (37 °C, 5% CO2). After 24 h of cell seeding, control and protein agents were added to each well. After 24 h in the incubator, 10 μL of CCK8 solution was added to each well, and the culture plate was incubated in the incubator for 0.5 - 2 hours. The absorbance at 450 nm was measured using an enzyme-linked immunosorbent assay (ELISA) reader.

[0155] GhJAZ1 has strong interactions with the proto-oncogenes c-Myc and NMYC. After constructing a prokaryotic expression plasmid of GhJAZ1 with a His tag to express the protein (see step 2 of Example 4), WB detection found that the GhJAZ1 protein with 6 His tags was approximately 30 KD in size ( Figure 7In A), breast cancer cells (MCF7) and mammary epithelial cells (MCF10A) were treated with the purified GhJAZ1 protein, and cell viability was measured using CCK8. The results showed that as the concentration of the GhJAZ1 protein increased, the viability of MCF7 cells decreased significantly ( Figure 7 In B), there was no significant change in the viability of MCF10A cells ( Figure 7 In C).

[0156] Cell apoptosis staining method. Fluorescence microscopy detection: a. Inoculation and culture. Inoculate cells in multi-well plates such as 96-well plates, cell culture dishes or cell slides, and perform certain treatments on the cells according to the experimental design. b. Washing (optional). For adherent cells, aspirate the culture medium and wash the cells once with PBS; for suspension cells, centrifuge at 250 - 1000×g for 5 min at room temperature, aspirate the supernatant, and wash once with PBS. Phenol red or serum has certain interference with the detection of this kit. It is best to use a vacuum pump when aspirating the culture medium and PBS. If the residual liquid can be fully aspirated, PBS washing can be omitted. c. Staining. Add an appropriate volume of Calcein AM / PI detection working solution (Beyotime Biotechnology, Calcein / PI Cell Viability and Cytotoxicity Detection Kit, Cat. No.: C2015). Usually, add 100 μl to each well of a 96-well plate, 250 μl to each well of a 24-well plate, 500 μl to each well of a 12-well plate, and 1 ml to each well of a 6-well plate. Incubate at 37 °C in the dark for 30 min. The optimal incubation time varies for different cells. Taking 30 min as the initial incubation time, the staining time can be appropriately adjusted and optimized according to the actual staining effect later to obtain a more ideal staining effect. d. Detection. After the incubation, observe the staining effect under a fluorescence microscope (Calcein AM is green fluorescence, Ex / Em = 494 / 517 nm; PI is red fluorescence, Ex / Em = 535 / 617 nm). Note that the entire process needs to be carried out under light protection.

[0157] Apoptosis staining was performed on the cells, and the results showed that the mortality rate of MCF7 cells increased significantly after treatment with the GhJAZ1 protein ( Figure 7 In D - K). We further treated lung cancer cells A549, ovarian cancer cells SKOV3, and cervical cancer cells Hela with the GhJAZ1 protein, and found that these cancer cells could all be inhibited by the GhJAZ1 protein ( Figure 7 In L - N). These experimental results indicate that the GhJAZ1 protein can inhibit cancer cells efficiently and broadly.

[0158] 4. Analysis of the effects of the GhJAZ1 protein on the cell cycle and apoptosis of tumor cells

[0159] To further explore the molecular mechanism of GhJAZ1 protein in inhibiting tumor cells, apoptosis and cell cycle were detected 24 hours after treating MCF7 breast cancer cells and normal mammary epithelial cells MCF10A with GhJAZ1 protein (using the IC50 concentration, i.e., 0.691 μM).

[0160] Apoptosis detection: Collect the cells in a 1.5 EP tube, centrifuge at 300 g for 5 min, discard the supernatant, collect the cells, wash with PBS and resuspend. Take 1×10 5 resuspended cells, centrifuge at 300 g for 5 min, discard the supernatant. Wash with PBS, add 500 μl of diluted 1×Annexin V Binding Buffer working solution to resuspend the cells, add 5 μL of Annexin V-FITC and 5 μL of propidium iodide (PI) staining solution, vortex and mix well, then incubate in the dark at room temperature for 15 - 20 min, and detect by machine. Specifically, use the Annexin V-FITC / PI fluorescence double staining cell apoptosis detection kit (Wuhan Punosai Life Technology Co., Ltd., product number: P-CA-201).

[0161] Cell cycle detection: Digest the cells moderately with trypsin, centrifuge to collect the cells, remove the supernatant, wash the cells with PBS, centrifuge at 1500 rpm for 5 min to collect, and prepare a single cell suspension (concentration: 1×10 6 / ml). Then take 1 ml of the single cell suspension and centrifuge, discard the supernatant, add 500 μl of pre-cooled 70% ethanol to fix the cells, and fix overnight at 4°C. The next day, centrifuge to collect the cells, resuspend with 1 ml of PBS and centrifuge, add 100 μl of RNase A solution to the cell pellet to resuspend, incubate in a water bath at 37°C for 30 min, add 400 μl of PI staining solution and mix well, incubate in the dark at 4°C for 30 min, and detect by machine, and record the red fluorescence at the excitation wavelength of 488 nm. Specifically, use the DNA content detection kit (cell cycle) (Solarbio, product number: CA1510).

[0162] The apoptosis detection results showed that GhJAZ1 led to an increased apoptosis rate of cancer cells ( Figure 8 in A - C), while having no significant effect on normal cells ( Figure 8 in D - F). The cell cycle detection results showed that treatment with GhJAZ1 led to a decrease in the proportion of cancer cells in the S phase, an increase in the proportion of cells in the G2 - M phase, and cell cycle arrest in the G2 - M phase ( Figure 8 in G - I), while there was no G2 - M phase arrest in MCF10A cells ( Figure 8 in J - L).

[0163] 5. Regulation of GhJAZ1 on c-Myc downstream genes

[0164] To further explore the mechanism by which GhJAZ1 inhibits cancer cell proliferation, we performed transcriptome sequencing on MCF7 cells treated with GhJAZ1 protein (using the IC50 concentration, i.e., 0.691 μM) for 24 h. The results showed that the expression of cell cycle-related genes was downregulated ( Figure 9 in A), and RT-PCR (the method steps were the same as in Example 1) verification was consistent with the transcriptome results ( Figure 9 in B). Among them, the RT-PCR primer sequences are as follows:

[0165] For the internal reference gene Aactin, the detection primers are:

[0166] Human-actin-F: 5-CATGTACGTTGCTATCCAGGC-3;

[0167] Human-actin-R: 5-CTCCTTAATGTCACGCACGAT-3.

[0168] The primers used to detect DHFR are:

[0169] Q-DHFR-F: 5-ATGGTTGGTTCGCTAAACTGCATC-3;

[0170] Q-DHFR-R: 5-CCTTGAGTTCTCTGCTGAGAACTAAATTAA-3.

[0171] The primers used to detect CDC45 are:

[0172] Q-CDC45-F: 5-CTTTGACTACGAGCAGTATGAATATCATG-3;

[0173] Q-CDC45-R: 5-TGTGCAGTCCACGGAGAGTGTG-3.

[0174] The primers used to detect CDCA7 are:

[0175] Q-CDCA7-F: 5-CTGATGACAGTTGTGACAGCTTTGC-3;

[0176] Q-CDCA7-R: 5-GAATCGGAGTTGGAATCAGTCACA-3.

[0177] The primers used to detect CDC16 are:

[0178] Q-CDC16-F: 5-ATTGTATGAAGCATGTCGTTACCTTGC-3;

[0179] Q-CDC16-R: 5-TGTAGGTAGCCAGGGTTCGGTTAT-3.

[0180] The primers for detecting POLD1 are:

[0181] Q-POLD1-F: 5-AGGATGCCTACCTGCCACTG-3;

[0182] Q-POLD1-R: 5-GGTACAGCGAGGAGAAGTCCAGG-3.

[0183] The primers for detecting POLK are:

[0184] Q-POLK-F: 5-AGACAAGCTGTGATGGACTTCATCAA-3;

[0185] Q-POLK-R: 5-CTCTTCCGCTTTATTTATCTCACTGAATG-3.

[0186] According to the c-Myc downstream genes reported in the literature, further verification was carried out by RT-PCR. The results showed that in cancer cells MCF7, after treatment with GhJAZ1, the apoptosis-related genes Bcl2, BAX, HK2, FASN, MCM5 downstream of c-Myc were significantly down-regulated, and the P21 gene was significantly up-regulated; the cell cycle-related genes CCNA2, CCNB1, CCND1 were significantly down-regulated, and CCNE1 was significantly up-regulated ( Figure 9 in C). In normal cells MCF10A, only Bcl2 and MCM5 decreased significantly, but the decrease amplitude was low, and P21C, CNB1, CCND1, and CCNE1 all increased significantly, but the increase amplitude was also small, and there was no obvious overall trend ( Figure 9 in D). Among them, the primer sequences are:

[0187] For the internal reference gene Aactin, the detection primers are:

[0188] Human-actin-F: 5-CATGTACGTTGCTATCCAGGC-3;

[0189] Human-actin-R: 5-CTCCTTAATGTCACGCACGAT-3.

[0190] The primers for detecting c-MYC are:

[0191] Q-c-MYC-F: 5-ATGGTGACCGAGCTGCTGGG-3;

[0192] Q-c-MYC-R: 5'-CCACCGAGGGGTCGATGCA-3'.

[0193] The primers for detecting P21 are:

[0194] Q-P21-F: 5'-GATGTCCGTCAGAACCCATGCG-3';

[0195] Q-P21-R: 5'-CGCTCCCAGGCGAAGTCACC-3'.

[0196] The primers for detecting Bcl2 are:

[0197] Q-Bcl2-F: 5'-GGGTACGATAACCGGGAGATAGTGATG-3';

[0198] Q-Bcl2-R: 5'-GCACCGGGCTGAGCGCAG-3'.

[0199] The primers for detecting BAX are:

[0200] Q-BAX-F: 5'-GAGGCACCCGAGCTGGCC-3';

[0201] Q-BAX-R: 5'-TGATCAGTTCCGGCACCTTGGT-3'.

[0202] The primers for detecting HK2 are:

[0203] Q-HK2-F: 5'-CGCTGTGGTGGACAGGATACGAG-3';

[0204] Q-HK2-R: 5'-CTATCGCTGTCCAGCCTCACGGA-3'.

[0205] The primers for detecting FASN are:

[0206] Q-FASN-F: 5'-GGAAGCTGCCAGAGTCGGAGAACTT-3';

[0207] Q-FASN-R: 5'-GTCCACGATGGCTTCATAGGTGACTT-3'.

[0208] The primers for detecting MCM5 are:

[0209] Q-MCM5-F: 5'-ATGTCGGGATTCGACGATCCTG-3';

[0210] Q-MCM5-R: 5-TGTAATGCCGCTTGAGTTCATCCC-3.

[0211] The primers for detecting CCNA2 are:

[0212] Q-CCNA2-F: 5-CTATCCTCGTGGACTGGTTAGTTG-3;

[0213] Q-CCNA2-R: 5-AACTTTGAGGCTAACAGCATAGCA-3.

[0214] The primers for detecting CCNB1 are:

[0215] Q-CCNB1-F: 5-GAAATTCAGGTTGTTGCAGGAGAC-3;

[0216] Q-CCNB1-R: 5-GTGCTTAGTATAAGTGTTGTCAGTCACAAA-3.

[0217] The primers for detecting CCND1 are:

[0218] Q-CCND1-F: 5-AGCTCCTGTGCTGCGAAGTGGA-3;

[0219] Q-CCND1-R: 5-GACAGGAAGCGGTCCAGGTAGTTCA-3.

[0220] The primers for detecting CCNE1 are:

[0221] Q-CCNE1-F: 5-TTTACCCAAACTCAACGTGCAAG-3;

[0222] Q-CCNE1-R: 5-TAGACTTCACACACCTCCATTAACCA-3.

[0223] Example 5. Analysis of the entry of GhJAZ1 protein into cancer cells and cell nuclei

[0224] 1. Screening and verification of the interaction between GhJAZ1 and tumor cell surface receptor proteins

[0225] To explore how GhJAZ1 enters cancer cells, we cloned most of the tumor surface receptor genes, including uPAR (NCBI database accession number NM_002659.4), CCK1R (NP_000721.1), CCK2R (NP_001350481.1), HER2 (NP_004439.2), GRPR (NP_005305.1), MUC1 (NP_001358649.1), VEGFR1 (NP_002010.2), Insulin receptor (NM_000208.4), NTR (XM_011528827.2), PSMA (XM_017017432.1), TFR1 (NM_001128148.3), TFR2 (NM_003227.4), PEPT1 (NM_005073.4), APN (NM_001150.3). Then, receptor proteins that interact with GhJAZ1 were screened by yeast two-hybrid (the experimental procedure is the same as in Example 3). The results showed that GhJAZ1 interacted with uPAR ( Figure 10 in A). Further COIP experiments (the method steps are the same as in Example 3) were carried out, and the results showed that GhJAZ1 interacted with uPAR in vivo ( Figure 10 in B). uPA is the ligand of uPAR. We detected the interaction between GhJAZ1 and uPA (NCBI database accession number NP_002649.2) and uPAR by yeast two-hybrid (the experimental procedure is the same as in Example 3). The results showed that GhJAZ1 interacted with the ATF domain of uPA, and the ATF domain of uPA also interacted with uPAR ( Figure 10 in C and D).

[0226] 2. Nuclear entry detection of GhJAZ1 protein

[0227] GhJAZ1 enters cells through the cancer cell surface receptor uPAR. Can it enter the nucleus? We further designed and constructed a prokaryotic expression vector, ligated GhJAZ1+RFP (amino acid sequence shown in SEQ ID No. 71, corresponding coding gene sequence such as SEQ ID No. 72) between NdeI and HindIII of the pET-30a(+) vector (Youbao Biotech, product number: VT1212), and prokaryotically expressed and purified the GhJAZ1+RFP protein with red fluorescence (the method steps are the same as in Step 2 of Example 4) ( Figure 11 [[ID=1�]]in A), and then treated MCF7 cells with the GhJAZ1+RFP protein and observed under a fluorescence microscope. The results showed that there was no light in the control under red fluorescence ( Figure 11In B-E), with the addition of IPR803 (uPAR inhibitor, concentration 50 μM) (MedChemExpress, catalog number: HY-111192), there is red fluorescence outside the cell nucleus. Figure 11 In F-I), the fluorescence of the added GhJAZ1+RFP protein (concentration 0.5 μM) shows a punctate distribution and coincides with the blue fluorescence of the cell nucleus. Figure 11 In J-M), it is proved that GhJAZ1 enters the cell nucleus.

[0228] Example 6. Analysis of the in vivo anti-tumor effect of GhJAZ1 on a mouse model of ovarian cancer

[0229] Human ovarian cancer cells SKOV3 were cultured. Before tumor inoculation, the cells were digested with trypsin, washed twice with PBS, and then resuspended in PBS to a concentration of 1×10 8 cells / mL. The cell suspension was injected subcutaneously into the axilla of nude mice, and the inoculation amount for each nude mouse was 0.2 mL. After inoculation, the nude mice were randomly divided into 2 groups, namely the PBS group and the GhJAZ1 protein (5 mg / kg) treatment group, with 8 mice in each group. After tumor inoculation, when the tumor volume reached about 50 - 100 mm 3 treatment began. The drug was injected into the tumor once every 2 days. Before each drug administration, the length and width of the tumor were measured. The drug administration cycle was 14 days, 7 times. During the whole experiment, the body weight of the nude mice was monitored while administering the drug. The tumor size was measured using a vernier caliper, and the tumor volume was calculated using the formula V = 1 / 2 × length × width × width, and the tumor growth curve was plotted. After 14 days, the tumors were taken out, photographed, and weighed.

[0230] The results of intratumoral injection showed that as the injection days were delayed, the tumor volume of the mice in the GhJAZ1 protein injection group was significantly lower than that of the control group. Figure 12 In B), Figure 12 A and C in the figure show the mice and the removed tumors 14 days after injection. The weight of the tumors in the injection group was extremely significantly lower than that of the control group. Figure 12 In D).

[0231] Example 7. Analyzing the interaction domains between GhJAZ1 and uPAR, c-Myc by yeast two-hybrid

[0232] Using the yeast two-hybrid competent cells Y2HGold from Shanghai Weidi Company, constructs PGBKT7::GhJAZ1, PGBKT7::GhJAZ1 ΔNT 、PGBKT7::GhJAZ1 ΔZIM 、PGBKT7::GhJAZ1 ΔJasEqual cotton JAZ yeast plasmids (see Example 3) and vector plasmids such as PGADT7::uPAR (NCBI database accession number of uPAR: NM_002659.4), PGADT7::uPAR-D1D2 (coding gene sequence of uPAR-D1D2 is as shown in SEQ ID No. 80, corresponding amino acid sequence is as shown in SEQ ID No. 79), PGADT7::uPAR-D1HD3 (coding gene sequence of uPAR-D1HD3 is as shown in SEQ ID No. 82, corresponding amino acid sequence is as shown in SEQ ID No. 81), PGADT7::uPAR-D2D3 (coding gene sequence of uPAR-D2D3 is as shown in SEQ ID No. 84, corresponding amino acid sequence is as shown in SEQ ID No. 83), PGADT7::c-Myc (NCBI accession number of c-Myc: NM_002467.6), PGADT7::NMyc (NCBI accession number of NMyc: NP_005369.2), PGADT7::c-Myc-150aa (coding gene sequence of c-Myc-150aa is as shown in SEQ ID No. 86, corresponding amino acid sequence is as shown in SEQ ID No. 85), PGADT7::c-Myc-215aa (coding gene sequence of c-Myc-215aa is as shown in SEQ ID No. 88, corresponding amino acid sequence is as shown in SEQ ID No. 87) (the CDS of the above genes was subcloned between the EcoRI and BamHI restriction sites of pGADT7 (Youbao Biotech, VT1639) or pGBKT7 (Youbao Biotech, VT16398)) were co-transformed into yeast competent cells respectively, and then the experiment was carried out according to the competent cell instruction manual (the method steps are the same as Example 3) to detect the interaction domains between cotton GhJAZ1 and uPAR, c-Myc.

[0233] The yeast two-hybrid results showed that the interaction between GhJAZ1 lacking the NT domain and uPAR was significantly weakened ( Figure 13 in A); the interaction between uPAR lacking the D3 domain and GhJAZ1 was significantly weakened to none ( Figure 13 in B), indicating that the NT domain of GhJAZ1 and the D3 domain of uPAR are involved in the interaction. The interaction between GhJAZ1 lacking the NT domain and c-Myc was significantly weakened ( Figure 13 in C); there were strong interactions between GhJAZ1 and c-Myc truncated at the first 150 amino acids and the first 215 amino acids ( Figure 13 in D and E), indicating that the NT domain of GhJAZ1 interacts with the N-terminus of c-Myc.

[0234] The present invention has been described in detail above. For those skilled in the art, without departing from the gist and scope of the present invention and without the need for unnecessary experiments, the present invention can be implemented within a relatively wide range under equivalent parameters, concentrations, and conditions. Although specific embodiments of the present invention are given, it should be understood that the present invention can be further improved. In short, according to the principle of the present invention, this application intends to cover any modifications, uses, or improvements to the present invention, including those that depart from the scope disclosed in this application but are made using conventional techniques known in the art. Some basic features can be applied within the scope of the following appended claims. <110> Institute of Cotton Research, Chinese Academy of Agricultural Sciences <120> Application of Plant JAZ Protein and Derived Polypeptides in Preventing and Treating Tumors in Humans and Animals <130> GNCLN211750 <160> 89 <170> PatentIn version 3.5 <210> 1 <211> 209 <212> PRT <213> Artificial sequence <400> 1 Met Ser Arg Ala Thr Val Glu Leu Asp Phe Phe Gly Met Glu Lys Ala 1 5 10 15 Asn Ser Cys Lys Ser Gln Phe Gln Lys Phe Leu Asp Arg Arg Arg Ser 20 25 30 Phe Arg Gly Ile Gln Gly Ala Ile Ser Lys Met Asn Pro Glu Leu Ile 35 40 45 Lys Ser Val Ile Ala Ser Gly Ser Thr Thr Arg Asn Pro Val Asp Trp 50 55 60 Arg Lys Ser Phe Ser Val Pro Ser Ser Pro Lys Glu Asp Arg Ser Thr 65 70 75 80 Ser Leu Pro Ser Leu Pro Leu Leu Asn Pro Ala Leu Arg Ser Ile Pro 85 90 95 Ser Glu Glu Ser Pro Glu Thr Ala Pro Leu Thr Ile Phe Tyr Asn Gly 100 105 110 Thr Val Ser Val Ile Asn Val Pro Arg Asp Lys Val Arg Gly Ser Ile 115 120 125 Phe Lys Leu Ala Val Glu Gly Ser Ser Lys Asn Ile Glu Ser Val Asp 130 135 140 Ser Ser Lys Ala Ala Asn Pro Ser Ser Asp Gln Gln Asn Leu Leu Glu 145 150 155 160 Ala Arg Asn Gly Asp Leu Pro Ile Ala Arg Arg Lys Ser Leu Gln Arg 165 170 175 Phe Leu Glu Lys Arg Lys Glu Arg Leu Asp Ile Leu Leu Cys Trp Ile 180 185 190 Arg Ile Tyr Arg Ile Phe Thr Asn Phe Leu Lys Lys Lys Lys Asn Asn 195 200 205 Asn <210> 2 <211> 363 <212> PRT <213> Artificial sequence <400> 2 Met Glu Arg Asp Phe Leu Gly Leu Asn Ser Asn Glu Pro Leu Ala Val 1 5 10 15 Val Lys Asp Asp Val Asn Thr Asp Lys Tyr Lys Gln Ile Gly Phe Thr 20 25 30 Lys Ser Ser Gly Ile Gln Trp Pro Phe Ser Asn Lys Val Phe Ala Val 35 40 45 Pro Gln Leu Met Asn Phe Asn Phe Ala Gln Gly Asp Lys Thr Lys Lys 50 55 60 Thr Gly Tyr Asp Ser Lys Val Ser Pro Leu Phe Met Pro Ile Ser Thr 65 70 75 80 Met Asp Ala Ser Glu Leu Gln Lys Ser Phe Asn His Asn Trp Asn Gly 85 90 95 Gly Gly His Phe Ser Leu Thr Asp Ser His Val Gln His Asn Thr Asn 100 105 110 Met Phe Pro Ala Ser Asn Gln Thr Ile Ser Val Ser Gly Ser Asn Pro 115 120 125 Phe Val Lys Asn His Phe Thr Thr Thr Gly Gln Asn Phe Pro Ala Asn 130 135 140 Thr Ile Lys Pro Gln Phe Leu Gly Gly Val Pro Val Thr Thr Pro His 145 150 155 160 Ser Val Leu Pro Thr Leu Gly Thr Val Gly Gly Ser Val Glu Pro Cys 165 170 175 Ala Gln Thr Ser Gly Ser Pro Ala Gln Leu Thr Ile Phe Tyr Ala Gly 180 185 190 Glu Val Asn Val Phe Asp Asp Ile Thr Pro Glu Lys Ala Gln Ala Ile 195 200 205 Met Phe Leu Ala Gly Asn Gly Ser Ser Met Ala Ser Asn Ser Ala Tyr 210 215 220 Pro Lys Pro Pro Val Gln Thr Pro Ile Leu Lys Pro Val Gln Val Asp 225 230 235 240 Ser Val Pro Ala Asn Gln Leu Ile Asn Thr Gln Leu Ser Phe Gly Met 245 250 255 Pro Ser Pro Leu Pro Val Ser Ser His Ala Gly Met Gln Ser Trp Ser 260 265 270 Gly Ser Thr Ser Thr Glu Glu Gln Ile Ile Cys Lys Ala Ser Val Pro 275 280 285 Pro Thr Pro Ser Thr Pro Ile Ser Lys Leu Glu Ser Pro Asn Leu Val 290 295 300 Asn Thr Met Gly Ser Asp Ala Ala Thr Gly Met Met Pro Ser Val Pro 305 310 315 320 Gln Ala Arg Lys Ala Ser Leu Ala Arg Phe Leu Glu Lys Arg Lys Gly 325 330 335 Arg Ile Met Ser Thr Ala Ser Pro Tyr Asn Leu Ile Ser Lys Lys Ser 340 345 350 Leu Asp Tyr Ala Thr Thr Met Glu Ser Asn Ala 355 360 <210> 3 <211> 197 <212> PRT <213> Artificial sequence <400> 3 Met Ser Arg Ala Ser Val Glu Leu Asp Phe Phe Gly Met Glu Lys Gln 1 5 10 15 Asn Phe Cys Lys Ser Arg Phe Gln Lys Ser Leu Asp Arg Arg Leu Ser 20 25 30 Phe Arg Gly Leu Gln Gly Ala Leu Ser Lys Val Asn Pro Glu Leu Ile 35 40 45 Lys Thr Val Ile Ala Ser Ser Leu Lys Asn Pro Gln Gly Gln Asp Asn 50 55 60 Val Tyr Gln Met Asp Ser Asn Lys Ser Phe Ser Val Pro Ser Ser Pro 65 70 75 80 Lys Glu Thr Gln Ser Leu Phe Pro Ala Leu Pro Leu Leu Asn Pro Ala 85 90 95 Ala Arg Ala Thr Ser Glu Asn Gly Pro Glu Ile Ala Pro Leu Thr Ile 100 105 110 Phe Tyr Asn Gly Thr Val Ser Val Phe Asn Val Pro Arg Asp Lys Ala 115 120 125 Glu Ser Ile Leu Lys Leu Ala Val Glu Val Glu Gly Asn Ser Lys Asn 130 135 140 Val Glu Pro Ile Asp Ser Lys Val Ala Ser Pro Pro Ser Asp Arg Gln 145 150 155 160 Gln Leu Leu Glu Thr Leu Asn Gly Asp Leu Pro Ile Ala Arg Arg Lys 165 170 175 Ser Leu Gln Arg Phe Leu Glu Lys Arg Lys Glu Arg Met Thr Cys Ala 180 185 190 Ser Pro Tyr Ala Cys 195 <210> 4 <211> 240 <212> PRT <213> Artificial sequence <400> 4 Met Ser Ser Cys Ser Glu Ser Thr Ala Met Lys Pro Ala Arg Ser Pro 1 5 10 15 Glu Lys Pro Ser Phe Ala Gln Thr Cys Asn Leu Leu Ser Gln Tyr Leu 20 25 30 Lys Glu Lys Gly Ser Phe Gly Asp Leu Ser Leu Gly Met Thr Cys Asn 35 40 45 Val Glu Ala Asn Gly Thr Pro Val Val Pro Pro Pro Thr Met Asn Leu 50 55 60 Phe Pro Leu Asn Asp Lys Ser Asp Asp Val Cys Gly Arg Asn Gly Gly 65 70 75 80 Asn Pro Lys Asn Leu Thr Ser Met Asp Leu Phe Pro Gln Gln Ala Gly 85 90 95 Leu Ala Ser Lys Asp Asp Ser Pro Asn Lys Leu Glu Pro Gln Thr Ala 100 105 110 Pro Met Thr Ile Phe Tyr Gly Gly Gln Val Ile Val Phe Asn Asp Phe 115 120 125 Pro Ala Asn Lys Ala Lys Glu Ile Met Leu Leu Ala Ser Asn Ser Ser 130 135 140 Ser Gln Ser Asn Asn Ser Phe Asn Pro Ile Pro Phe Thr Ser Ser Ile 145 150 155 160 Ala Arg Ser Pro Ile Glu Ser Ser Ile Gly Val Pro Pro Thr Ser Lys 165 170 175 Pro Val His Pro Ala Gln Arg Ala Val Pro Gly Asp Leu Pro Ile Ala 180 185 190 Arg Arg Ala Ser Leu His Arg Phe Leu Glu Lys Arg Lys Asp Arg Ile 195 200 205 Thr Ala Lys Ala Pro Tyr Gln Ile Asn Asn Ser Ala Ala Ala Pro Ser 210 215 220 Leu Ser Gly Asp Asn Lys Ser Trp Leu Gly Leu Ala Ala Gln Ser Pro 225 230 235 240 <210> 5 <211> 216 <212> PRT <213> Artificial sequence <400> 5 Met Glu Gly Glu Ala Gly Ser Tyr Glu Asp Val Lys Pro Asn Val Val 1 5 10 15 Val Lys Gln Ser Asn Gly Asp Val Val Gly Asp Asn Gly Val Gly Asn 20 25 30 Leu Gly Ser Val Asp Ala Pro Asp Phe Leu Ser Lys Lys Asn Phe Gln 35 40 45 Asn Cys Pro Ala Pro Ser Gln Leu Thr Ile Phe Tyr Asp Gly His Val 50 55 60 Cys Val Phe Asp Ala Ile Pro Val Glu Lys Val Arg Glu Ile Met Leu 65 70 75 80 Ile Ala Ala Thr Ala Ala Ala Gly Ala Ala Asn Ser Val Asp Met Lys 85 90 95 Lys Val Ala Thr Asp Cys Ala Thr Thr Ser Pro Val Leu Thr Arg Ser 100 105 110 Pro Ser Leu Gln Ser Thr Ala Thr Ala Thr Ala Leu Ala Ser Pro Gln 115 120 125 Ala Gln Val Tyr Pro Ile Asn Arg Thr Pro Phe Cys Lys Leu Lys Glu 130 135 140 Leu Pro Ile Ala Arg Arg His Ser Leu Gln Arg Phe Phe Glu Lys Arg 145 150 155 160 Arg Asp Arg Leu Val Asn Arg Asn Pro Tyr Pro Asn Pro Ser Thr Pro 165 170 175 Lys Ser Phe Asp Asp Thr Lys Ala Asn Leu Ser Ala Ala Thr Ser Pro 180 185 190 Glu Ser Gly Cys Phe Gly Lys Ser Pro Val Ala Gln Glu Glu Phe His 195 200 205 Pro Lys Ala Pro Ala His Val Ala 210 215 <210> 6 <211> 226 <212> PRT <213> Artificial sequence <400> 6 Met Thr Lys Asn Phe Leu Pro Asn Met Glu Asn Ser Tyr Glu Ser Leu 1 5 10 15 Lys Pro Asn Leu Gly Ala Ser Thr Ser Lys Ser Asn Ala Lys Arg Cys 20 25 30 Gly Phe Phe Pro Glu Ile Arg Ser Phe Gly Val Ser Ser Ser Lys Glu 35 40 45 Asp Thr Asn Arg Met Thr Asp Phe Arg Lys Pro Ala Lys Val Glu Pro 50 55 60 Lys Asn Ser Gln Met Thr Ile Phe Phe Gly Ser Gln Val Ala Val Phe 65 70 75 80 Asn Asp Phe Pro Ala Asp Lys Phe Lys Glu Ile Met Asp Leu Leu Pro 85 90 95 Ser His Gly Cys Ser Thr Ala Ser Gly Gly Val Val Asp Thr Val Met 100 105 110 Glu Lys Val Lys Ser Lys Ile Val Gln Ile Glu Pro Ser Asn His Glu 115 120 125 Ile Pro Asp Leu Asn Val Ala Thr Ala Thr Gly Asn Ser Pro Pro Pro 130 135 140 Pro His Asp Ser Ser Val Glu Trp His Gln Tyr Gly Gly Ser Gly Ser 145 150 155 160 Ser Asp Leu Arg Ile Ala Arg Arg Asn Ser Leu His Lys Phe Phe Glu 165 170 175 Lys Arg Lys Glu Arg Ala Thr Ala Arg Ala Pro Tyr Gln Val Asn Asn 180 185 190 Ala Arg Gly Ser Thr Pro Pro Pro Lys Pro Asn Glu Asn Lys Ser Ser 195 200 205 His Glu Glu Gly Gln Ser Ser Lys Glu Ala Ser Arg Asp Leu Asp Leu 210 215 220 Lys Leu 225 <210> 7 <211> 119 <212> PRT <213> Artificial sequence <400> 7 Met Arg Arg Asn Cys Asn Leu Glu Leu Arg Leu Leu Pro Ser Ser Tyr 1 5 10 15 Pro Ser Asp Ser His Asp Met Met Glu Glu Arg Ile Glu Ser Pro Lys 20 25 30 Thr Gln Gln Gln Gln Leu Thr Ile Phe Tyr Asn Gly Arg Val Cys Val 35 40 45 Ser Asp Val Thr Glu Leu Gln Ala Lys Ala Ile Leu Met Leu Ala Asn 50 55 60 Arg Glu Arg Asp Glu Arg Met Lys Ser Pro Thr Gly Trp Glu Pro Val 65 70 75 80 Ser Pro Thr Leu Lys Ser Gln Val Lys Cys Pro Asn Thr Ala Leu Ser 85 90 95 Met Lys Arg Ser Leu Gln Arg Phe Leu Gln Lys Arg Lys Thr Arg Ile 100 105 110 Gln Ala Thr Ser Pro Tyr His 115 <210> 8 <211> 364 <212> PRT <213> Artificial sequence <400> 8 Met Glu Arg Asp Phe Leu Gly Leu Asn Ser Lys Gln Ser Phe Pro Leu 1 5 10 15 Val Lys Glu Glu Val Glu Glu Ile Gly Phe Thr Lys Ser Leu Gly Ile 20 25 30 Gln Trp Pro Ile Ser Asn Lys Val Ser Ser Ala Val Pro Pro Gln Gln 35 40 45 Met Ser Phe Asp Phe Ala Gln Val Asp Asn Ala Lys Arg Ile Gly Tyr 50 55 60 Asp Ser Ile Val Ser Pro Ala Phe Met His Ile Ser Pro Pro Asp Ala 65 70 75 80 Ala Gln Leu Gln Lys Ser Phe Asn His Asn Arg Gln Arg Val Gly Asn 85 90 95 His Phe Pro Phe Thr Ala Ala Ser Val Gln His Asp Ala His His Val 100 105 110 Gln Arg Pro Tyr Asp Met Lys Met Phe Pro Val Ser Asn Gln Ser Val 115 120 125 Pro Val Ser Thr Thr Asn Pro Phe Met Asn Asn His Phe Thr Thr Thr 130 135 140 Ala Met Lys Ser Gln Leu Leu Gly Gly Ile Pro Val Thr Ser Pro Pro 145 150 155 160 His Ser Val Leu Pro Thr Leu Ser Ser Phe Gly Gly Ser Ile Glu Pro 165 170 175 Arg Lys Ser Val Arg Gly Leu Gly Asp Ser Arg Ser Pro Val Gln Leu 180 185 190 Thr Ile Phe Tyr Ala Gly Thr Val Asn Val Tyr Asp Asp Ile Thr Pro 195 200 205 Glu Lys Ala Gln Ala Ile Met Leu Leu Ala Gly Asn Gly Ser Ser Leu 210 215 220 Thr Ser Asn Val Ala His Pro Lys Val Gln Val Gln Ala Pro Ile Ser 225 230 235 240 Lys Pro Leu Gln Phe Glu Asn Leu Pro Thr Asn His Phe Thr Asn Glu 245 250 255 Gln Leu Cys Ser Gly Ile Pro Ser Pro Leu Ser Val Ser Ser His Thr 260 265 270 Gly Val Gln Ser Arg Ser Gly Ser Thr Ser Thr Asp Glu Lys Thr Val 275 280 285 Cys Lys Thr Thr Gly Ser Leu Thr Thr Pro Ile Ser Leu Val Glu Ser 290 295 300 Pro Lys Leu Ala Asn Thr Met Gly Pro Val Thr Pro Thr Ser Ile Met 305 310 315 320 Pro Thr Val Pro Gln Ala Arg Lys Ala Ser Leu Ala Arg Phe Leu Glu 325 330 335 Lys Arg Lys Glu Arg Ile Met Thr Ala Ser Pro Tyr Asp Leu Ser Lys 340 345 350 Lys Pro Pro His Cys Thr Thr Gln Gly Ser Asn Ala 355 360 <210> 9 <211> 263 <212> PRT <213> Artificial sequence <400> 9 Met Asn Met Ser Cys Ser Pro Glu Phe Leu Val Gln Lys Pro Thr Arg 1 5 10 15 Ser Pro Glu Lys Thr Ser Phe Thr Gln Thr Cys Asn Leu Leu Ser Gln 20 25 30 Tyr Leu Lys Glu Lys Gly Ser Phe Gly Asp Leu Ser Leu Gly Met Thr 35 40 45 Cys Asn Asp Glu Ala Asn Glu Thr Pro Glu Met Leu Arg Pro Thr Met 50 55 60 Asn Leu Phe Pro Val Asn Glu Lys Ser Gly Asp Asp Cys Leu Ala Ala 65 70 75 80 Pro Pro Pro Arg Lys Leu Arg Ser Met Asp Leu Phe Pro Asn Gln Ala 85 90 95 Ala Phe Ser Ser Pro Lys Asp Asp Ala Leu Lys Ser Thr Met Asn Lys 100 105 110 Leu Gly Ser Ser Val Glu Pro Gln Thr Ala Gln Met Thr Ile Phe Tyr 115 120 125 Gly Gly Gln Val Ile Val Phe Asn Asp Phe Pro Ala Asp Lys Ala Lys 130 135 140 Glu Ile Met Leu Leu Ala Gly Lys Gly Ser Ser Gln Ser Asn Ser Phe 145 150 155 160 Asn Thr Asn Pro Pro His Ile Asn Ala Pro Phe Thr Ser Thr Ile Ala 165 170 175 Thr Ser Pro Ile Glu Ser Gly Ile Gly Val Pro Pro Thr Pro Asn Phe 180 185 190 Ser Thr Thr Val Thr Gln Glu Cys Ile Arg Ser Ala Gln Arg Ser Ile 195 200 205 Pro Gly Asp Leu Pro Ile Ala Arg Arg Ala Ser Leu His Arg Phe Leu 210 215 220 Glu Lys Arg Lys Asp Arg Met Thr Thr Arg Ala Pro Tyr Glu Ile Ser 225 230 235 240 Asn Ser Thr Ala Ser Ser Ser Lys Pro Gly Asp Asp Lys Ser Trp Leu 245 250 255 Gly Leu Ala Ala Gln Ser Pro 260 <210> 10 <211> 228 <212> PRT <213> Artificial sequence <400> 10 Met Glu Glu Glu Ala Glu Ser Arg Glu Glu Val Lys Pro Asn Val Val 1 5 10 15 Val Lys Glu Thr Asn Gly Asp Ile Val Gly Asp Asn Asp Ala Gly Lys 20 25 30 Leu Gly Thr Ile Glu Thr Pro Asp Phe Leu Ser Gln Lys Ile Phe His 35 40 45 Asn Cys Ser Leu Pro Ile Leu Ala Ser Gly Val Ile Thr Thr Ser Pro 50 55 60 Ala Gln Ser Gln Leu Thr Ile Phe Tyr Gly Gly Lys Val Ser Val Phe 65 70 75 80 Asp Ala Ile Ser Ala Glu Lys Ile Gln Glu Ile Met Leu Ile Ala Ala 85 90 95 Thr Val Ala Ala Ala Asp Val Gly Ser Val Asp Met Lys Asn Ala Ala 100 105 110 Thr Asp Tyr Ala Thr Ile Ser Pro Ala Leu Thr Arg Cys Pro Ser Leu 115 120 125 Gln Ser Thr Ala Thr Ala Ser Ala Ser Pro Gln Ala Gln Leu Tyr Pro 130 135 140 Leu Pro Arg Thr Ser Phe Ser Lys Leu Gln Ala Glu Leu Pro Ile Ala 145 150 155 160 Arg Arg His Ser Leu Gln Arg Phe Leu Glu Lys Arg Arg Asp Arg Leu 165 170 175 Val Asn Lys Asn Pro Tyr Pro Gly Pro Ser Thr Pro Lys Met Ala Asp 180 185 190 Asp Val Lys Ala Asp Val Ser Ala Thr Thr Ser Pro Glu Ser Gly Tyr 195 200 205 Phe Lys Ala Ser Pro Ile Arg Gln Glu Asp Ile Gln Pro Lys Ala Pro 210 215 220 Ala His Val Ala 225 <210> 11 <211> 297 <212> PRT <213> Artificial sequence <400> 11 Met Ser Thr Gly Glu Met Val Ser Arg Ser Pro Leu Tyr Lys Pro Leu 1 5 10 15 Asn Gln Leu Thr Glu Asp Asp Ile Ser Gln Val Thr Arg Glu Asp Cys 20 25 30 Arg Leu Tyr Leu Lys Glu Lys Gly Met Arg Arg Pro Ser Trp Asn Lys 35 40 45 Ser Gln Ala Ile Gln Gln Val Ile Ser Leu Lys Thr Leu Leu Glu Thr 50 55 60 Thr Ser Asp Ser Glu Ala Val Glu Ala Ser Lys Lys Leu His Val Pro 65 70 75 80 Phe Pro Gln Asn Pro Pro Arg Phe Val Ser Asp Ser Thr Val Gln Pro 85 90 95 Asn Glu Thr Thr Arg His Lys Gly Ile Ser Val Pro Val Asn Glu Ser 100 105 110 Val Pro Arg Ile Arg Ser Asp Pro Ser Glu Phe Lys Phe Ser Arg Glu 115 120 125 Asn Ser Val Gln Thr Ala Val Ser Ala Asn Asp Ser Val Ser Pro Arg 130 135 140 Ser Ala Ser Val Ala Lys Glu Pro Ser Gly Gln Met Thr Ile Phe Tyr 145 150 155 160 Cys Gly Lys Val Asn Val Tyr Asp Asn Ile Pro Gly Cys Lys Ala Glu 165 170 175 Ala Ile Leu Gln Phe Ala Ala Ser Pro Val Ser Phe Pro Gln Glu Thr 180 185 190 Leu Val Asp Gln Arg Thr Ser Pro Leu Ser Ile Pro Cys His Val Gln 195 200 205 Ala Ala Gly Asp Lys Val Ser Gln Arg Ser Pro Gly Val Ile Leu Ser 210 215 220 Ser Met Gln Ala Val Lys Val Ala Glu Asn Cys Arg Phe Pro Arg Asp 225 230 235 240 Asp Cys Asn Val Ser Tyr Glu Asp Ser Leu Glu Gly Pro Thr Ser Arg 245 250 255 Asn Ala Leu Leu Gln Arg Tyr Leu Glu Lys Lys Lys Asp Arg Tyr Ile 260 265 270 Arg Ser Met Met Leu Asp Gln His Ser Ala Ile Ile Ala His Val Asp 275 280 285 Met Asp Gly Ala Gly Val Pro Thr Glu 290 295 <210> 12 <211> 252 <212> PRT <213> Artificial sequence <400> 12 Met Phe Gly Ser Pro Glu Tyr Thr Cys Leu Lys Pro Ala Ser Leu Pro 1 5 10 15 Glu Lys Pro Leu Phe Lys Arg Thr Cys Ser Leu Leu Ser Gln Tyr Leu 20 25 30 Lys Glu Lys Cys Ser Phe Gly Asp Leu Thr Leu Gly Ile Thr Cys Asn 35 40 45 Asn Asn Val Glu Lys Gly Met Pro Glu Ile Val Arg Pro Ala Thr Glu 50 55 60 Thr Thr Thr Thr Thr Met Asp Leu Phe Pro Arg Asp His Val Ser Gly 65 70 75 80 Val Met Arg Asn Ser Arg Ser Met Asp Leu Phe Pro Gln Gly Ala Gly 85 90 95 Phe Ser Ala Asp Asn Gly Ser Arg Arg Val Gly Val Asp Leu Glu Thr 100 105 110 Glu Asn Ala Ala Ala Pro Met Thr Ile Phe Tyr Cys Gly Gln Val Ile 115 120 125 Val Phe Asn Asp Phe Pro Ala Asp Lys Ala Lys Glu Ile Met Ala Leu 130 135 140 Ala Ser Lys Cys Ser Ser Glu Asn Pro Lys Thr Asn Thr Phe Val Pro 145 150 155 160 Gly Ser Pro Asn Glu Ser Gly Leu Gly Val His Ser Asn Ser Asn Asp 165 170 175 Gln Val Pro Asn Ile Arg Thr Asn Val Ser Thr Ser Arg Glu Cys Val 180 185 190 Arg Ser Ile Pro Gly Asp Leu Pro Ile Ala Arg Arg Ala Ser Leu His 195 200 205 Arg Phe Leu Glu Lys Arg Lys Asp Arg Ile Thr Ser Lys Ala Pro Tyr 210 215 220 Pro Ile Asn Gly Ser Ala Gly Ala Ser Pro Pro Lys Pro Gly Asn Ser 225 230 235 240 Lys Pro Trp Leu Gly Leu Ala Val Glu Ser Leu Gln 245 250 <210> 13 <211> 125 <212> PRT <213> Artificial sequence <400> 13 Met Arg Arg Asn Cys Asn Leu Glu Leu Glu Leu Phe Pro Ser Arg Met 1 5 10 15 Phe Pro Gly His Arg Gln Asn Met Val Glu Glu Ser Lys Arg Ser Pro 20 25 30 Gln Asn Gln Gln Gln Gln Leu Thr Ile Phe Tyr Asn Gly Arg Val Cys 35 40 45 Val Cys Asp Ile Thr Glu Leu Gln Ala Arg Ala Ile Leu Met Arg Ala 50 55 60 Asn Gln Glu Thr Asp Glu Arg Ile Lys Thr Pro Thr Gly Ser Glu Pro 65 70 75 80 Asp Ser Pro Thr Ser Ser Ser Ser Thr Ser Leu Ser Arg Leu Cys Ser 85 90 95 Pro Asn Ala Gly Leu Ser Met Lys Lys Ser Leu Gln Arg Phe Leu Gln 100 105 110 Lys Arg Lys Asn Arg Ile Gln Ala Thr Ser Pro Tyr His 115 120 125 <210> 14 <211> 120 <212> PRT <213> Artificial sequence <400> 14 Met Arg Arg Asn Cys Asn Leu Glu Leu Arg Leu His Pro Ser Ser Tyr 1 5 10 15 Ser Gly Gly Asp Arg Val Glu Glu Ser Ser Glu Asn Pro Glu Asn Gln 20 25 30 Gln Gln Gln Leu Thr Ile Phe Tyr Asn Gly Arg Val Cys Val Cys Asp 35 40 45 Val Thr Glu Ile Gln Ala Arg Ala Ile Leu Met Ile Ala Asn Arg Glu 50 55 60 Thr Asp Glu Arg Leu Arg Thr Pro Arg Gln Gly Ser Glu Pro Ala Ser 65 70 75 80 Pro Met Val His Ser Gln Ile Asn Ser Pro Asn Asn Gly Leu Ser Thr 85 90 95 Ser Met Lys Arg Ser Leu Gln Arg Phe Leu Gln Lys Arg Lys Asn Arg 100 105 110 Ile Gln Ala Thr Ser Pro Tyr His 115 120 <210> 15 <211> 270 <212> PRT <213> Artificial sequence <400> 15 Met Ser Asn Leu Gly Gln Lys Ser Pro Asp Arg Ala Ser Phe Val Asn 1 5 10 15 Leu Leu Ser Gln Tyr Leu Lys Glu Lys Arg Asn Leu Gly Asp Phe Ser 20 25 30 Leu Gly Met Thr Ser Lys Pro Asp Ala Lys Gly Leu Glu Thr Ser Arg 35 40 45 Gln Gln Ala Lys Asn Met Asn Phe Leu Ser Asn Met Pro Asn Cys Ser 50 55 60 Glu Ser Ser Arg Pro Asn Leu Val Ala Ser Thr Ser Asn Val Lys Ser 65 70 75 80 Ser Asp Phe Phe Pro Glu Ile Gly Ser Phe Cys Ala Ser Ser Ser Lys 85 90 95 Glu Asp Thr Ile Asn Lys Thr Asp Phe Met Lys Ser Ala Ala Val Glu 100 105 110 Pro Lys Asn Ala Gln Leu Thr Ile Phe Phe Gly Gly Gln Val Leu Val 115 120 125 Tyr Asn Asp Phe Pro Ala Asp Lys Val Lys Glu Ile Met Ala Val Ala 130 135 140 Asn Arg Gly Trp Ser Thr Ala Cys Ser Gly Val Val Ala Asp Ser Ser 145 150 155 160 Met Glu Lys Leu Asn Ala Asn Leu Asp Lys Ile Asp Tyr Ser Ser Pro 165 170 175 His Ile Pro Asp Leu Asn Ile Thr Ser Ala Thr Val Asn Ser Ser Ala 180 185 190 Gln Asp Pro Ser Val Glu Arg His Gln Tyr Val Gly Ser Asp Leu Arg 195 200 205 Ile Ala Arg Arg Asn Ser Leu His Lys Phe Phe Glu Lys Arg Lys Asp 210 215 220 Arg Ala Ala Met Arg Ala Pro Tyr Gln Leu Ile Asn His Gln Gly Ser 225 230 235 240 Pro Pro Pro Pro Lys Pro Asp Glu Asn Lys Pro Ser His Glu Glu Gly 245 250 255 Gln Ser Ser Lys Glu Thr Pro Arg Asp Ile Asp Leu Asn Leu 260 265 270 <210> 16 <211> 337 <212> PRT <213> Artificial sequence <400> 16 Met Ser Pro Gly Glu Thr Val Ser Arg Ser Pro Leu Asp Lys Pro Leu 1 5 10 15 Asn Gln Leu Thr Glu Asp Asp Ile Ser Gln Val Thr Arg Glu Asp Cys 20 25 30 Arg Arg Tyr Leu Lys Glu Lys Gly Met Arg Arg Pro Ser Trp Asn Lys 35 40 45 Ser Gln Ala Ile Gln Gln Val Ile Ser Leu Lys Thr Leu Leu Glu Thr 50 55 60 Thr Ser Asp Ser Asp Ala Val Glu Ala Arg Lys Lys Leu Tyr Pro Pro 65 70 75 80 Cys Pro Glu Tyr Pro Pro Arg Val Val Ser Asp Ser Asn Val Leu Pro 85 90 95 Lys Glu Ile Thr Pro Asn Asn Gly Ile Leu Val Pro Val Pro Glu Ser 100 105 110 Val Pro Cys Pro His Ser Asn Pro Ser Lys Ser Asp Phe Ser Gly Asp 115 120 125 Asn Ser Gly Arg Thr Val Ile Ser Gly Asn Asp Ser Val Ser Pro Arg 130 135 140 Ile Ala Gly Ala Pro Lys Glu Ser Ala Gly Gln Met Thr Ile Phe Tyr 145 150 155 160 Cys Gly Lys Val Asn Val Tyr Asp Asp Met Pro Gly Cys Lys Ala Glu 165 170 175 Ala Ile Leu Gln Leu Ala Ala Ser Pro Val Ser Phe Pro His Glu Ile 180 185 190 Leu Ala Asp Gln Arg Thr Thr Pro Trp Ser Ile Pro Cys His Ser Gln 195 200 205 Ala Ala Ser Val Lys Thr Thr Pro Cys Ser Gln Met Val Ile Leu Pro 210 215 220 Pro Gln Gln Thr Glu Asn Cys Gln Phe Pro Arg Glu Glu Ser Asn Ala 225 230 235 240 Ser Leu Glu Asp Ser Leu Glu Gly Pro Thr Arg Arg Lys Ala Ser Val 245 250 255 Gln Arg Tyr Leu Glu Lys Lys Lys Asp Arg Phe Lys Asn Lys Arg Lys 260 265 270 Leu Ala Met Ser Ser Ser Pro Thr Leu Asp Ile Tyr Leu Asn Gln Val 275 280 285 Gly Asp Gln Phe Ser Asn Glu Gln Leu Lys Gln Ser Glu Pro Tyr Tyr 290 295 300 Ser Pro Gln Ala Glu Val Gln Arg Met Pro Leu Glu Cys Ser Ser Ile 305 310 315 320 Glu Asn Val Ala Lys Ile Pro Arg Leu Thr Thr Asp Gly Lys Gly Asp 325 330 335 Gln <210> 17 <211> 370 <212> PRT <213> Artificial sequence <400> 17 Met Glu Arg Asp Phe Ile Gly Leu Arg Ser Lys Asn Ala Ala Ile Thr 1 5 10 15 Ile Lys Val Glu Pro Ser Asn Ala Gln Pro Asp Asp Ser Val Leu Leu 20 25 30 Arg Gly Ser Gly Met Gln Leu Ser Phe Ser Asn Lys Val Ser Thr Val 35 40 45 Pro Gln Phe Leu Ser Phe Asp Gly Ala Arg Asp Asp Lys Pro Arg Lys 50 55 60 Ala Thr His Asp Thr Leu Leu Ser Ser Gly Phe Met Thr Ile Pro Ala 65 70 75 80 Ala Asp Ser Asn Lys Lys Pro His Pro Gly Leu Thr Gln Lys Gln Gly 85 90 95 Gly Asn His Tyr Ala Ala Thr Thr Tyr Gly Leu Gln Gln Leu Asp Gly 100 105 110 His Gln Ser Arg Leu Ser His Glu Ala Arg Ile Phe Pro Ser Ser Ser 115 120 125 Gln Pro Asn His Thr Ile Thr Val Ser Met Asn Thr Pro Leu Leu Gln 130 135 140 Pro His Leu Ala Ser Pro Gly Gln Asn Ile Ile Gly His Thr Ile Asn 145 150 155 160 Pro Gln Pro Phe Thr Gly Val Pro Ile Met Ala Ala Pro Val Ser Val 165 170 175 Val Pro Pro Ser Ser Pro Ile Ile Gly Thr Thr Asp Leu Arg Asn Ala 180 185 190 Ala Lys Ser Ser Arg Ala Pro Ala Gln Leu Thr Ile Phe Tyr Ala Gly 195 200 205 Ser Val Cys Val Tyr Asp Asp Val Ser Pro Asp Lys Ala Gln Ala Ile 210 215 220 Met Leu Leu Ala Gly Asn Val Ser Ser Thr Thr Gln Ser Lys Thr Ala 225 230 235 240 Pro Val Thr Gln Pro Gln Thr His Ile Pro Arg Pro Cys Thr Leu Ser 245 250 255 Pro Phe Ser Gly Leu Pro Asn His Leu Ser Val Thr Ser His Val Ser 260 265 270 Leu Gln Pro Val Ala Gly Ser Ser Gly Thr Asn Glu Leu Thr Ala Ala 275 280 285 Thr Arg Ile Gly Ala Leu Ala Ser Thr Asn Asn Gln Pro Asp Pro Pro 290 295 300 Lys Leu Val Asn Pro Ala Val Ala Val Pro Gln Ala Arg Lys Ala Ser 305 310 315 320 Leu Ala Arg Phe Leu Glu Lys Arg Lys Glu Arg Val Ser Asn Thr Ser 325 330 335 Pro Tyr Asn Ile Cys Lys Arg Ser Glu Ser Gly Pro Leu Ala Ser Asp 340 345 350 Gly Ile Ser Phe Ser Val Thr Ser Ala Gly Ser Ser Pro Leu Gln Ala 355 360 365 Ile Asn 370 <210> 18 <211> 206 <212> PRT <213> Artificial sequence <400> 18 Met Ser Arg Ala Thr Val Glu Leu Asp Phe Phe Gly Met Glu Lys Ala 1 5 10 15 Asn Ser Cys Lys Ser Gln Phe Gln Lys Phe Leu Asp Arg Arg Arg Ser 20 25 30 Phe Arg Gly Ile Gln Gly Ala Ile Ser Lys Met Asn Pro Glu Leu Ile 35 40 45 Lys Ser Val Ile Ala Ser Gly Ser Thr Asn Arg Asn Pro Val Asp Trp 50 55 60 Arg Lys Ser Phe Ser Val Pro Ser Ser Pro Lys Glu Asp Arg Ser Thr 65 70 75 80 Ser Leu Pro Ser Leu Pro Leu Leu Asn Pro Ala Leu Arg Ser Thr Pro 85 90 95 Ser Glu Asp Ser Pro Glu Thr Ala Pro Leu Thr Ile Phe Tyr Asn Gly 100 105 110 Thr Val Ser Val Ile Asn Val Pro Arg Asp Lys Ala Glu Ser Ile Phe 115 120 125 Lys Leu Ala Val Glu Gly Ser Ser Lys Asn Ile Glu Ser Val Asp Ser 130 135 140 Ser Lys Ala Ala Asn Pro Ser Ser Asp Gln Gln Asn Leu Leu Glu Ala 145 150 155 160 Arg Asn Gly Asp Leu Pro Ile Ala Arg Arg Lys Ser Leu Gln Arg Phe 165 170 175 Leu Glu Lys Arg Lys Glu Arg Leu Asp Ile Leu Leu Cys Trp Arg Arg 180 185 190 Ile Tyr Arg Ile Phe Thr Asn Phe Leu Lys Lys Asn Asn Asn 195 200 205 <210> 19 <211> 362 <212> PRT <213> Artificial sequence <400> 19 Met Glu Arg Asp Phe Leu Gly Leu Asn Ser Asn Glu Pro Leu Ala Val 1 5 10 15 Val Lys Asp Asp Val Asn Thr Asp Lys Tyr Lys Glu Ile Gly Phe Thr 20 25 30 Lys Ser Ser Gly Ile Gln Trp Pro Phe Ser Asn Lys Val Phe Ala Val 35 40 45 Pro Gln Leu Met Asn Phe Asn Phe Ala Gln Gly Asp Lys Thr Lys Lys 50 55 60 Thr Gly Tyr Asp Ser Lys Val Ser Pro Leu Phe Met Pro Ile Ser Thr 65 70 75 80 Met Asp Ala Ala Glu Leu Gln Lys Ser Phe Asn His Lys Trp Asn Gly 85 90 95 Gly Asn His Phe Ser Leu Thr Asp Ser His Val Gln His Asn Thr Asn 100 105 110 Met Phe Pro Ala Ser Asn Gln Thr Ile Ser Val Ser Gly Ser Asp Pro 115 120 125 Phe Val Lys Asn His Tyr Thr Thr Thr Gly Gln Lys Phe Pro Ala Asn 130 135 140 Thr Ile Lys Pro Gln Phe Phe Gly Gly Val Pro Val Thr Thr Pro His 145 150 155 160 Ser Val Leu Pro Thr Leu Gly Ser Val Gly Gly Ser Val Glu Pro Cys 165 170 175 Thr Lys Ala Ser Gly Ser Pro Ala Gln Leu Thr Ile Phe Tyr Ala Gly 180 185 190 Glu Val Asn Val Phe Asp Asp Ile Thr Pro Glu Lys Ala Gln Ala Ile 195 200 205 Methionine, Phenylalanine, Leucine, Alanine, Glycine, Asparagine, Glycine, Serine, Serine, Methionine, Alanine, Serine, Asparagine, Serine, Alanine, Tyrosine 210 215 220 Proline, Lysine, Proline, Proline, Valine, Glutamine, Threonine, Proline, Isoleucine, Leucine, Lysine, Proline, Valine, Glutamine, Valine, Aspartic acid 225 230 235 240 Serine, Valine, Proline, Alanine, Asparagine, Glutamine, Leucine, Isoleucine, Asparagine, Threonine, Glutamine, Leucine, Serine, Phenylalanine, Glycine, Lysine 245 250 255 Proline, Serine, Proline, Leucine, Proline, Valine, Serine, Serine, Histidine, Alanine, Glycine, Threonine, Glutamine, Serine, Tryptophan, Serine 260 265 270 Glycine, Serine, Threonine, Serine, Threonine, Glutamic acid, Glutamic acid, Glutamine, Isoleucine, Isoleucine, Cysteine, Lysine, Alanine, Serine, Valine, Proline 275 280 285 Serine, Threonine, Proline, Serine, Threonine, Proline, Isoleucine, Serine, Lysine, Leucine, Glutamic acid, Serine, Proline, Asparagine, Leucine, Valine 290 295 300 Asparagine, Threonine, Methionine, Glycine, Serine, Aspartic acid, Alanine, Alanine, Threonine, Glycine, Methionine, Methionine, Proline, Serine, Valine, Proline 305 310 315 320 Glutamine, Alanine, Arginine, Lysine, Alanine, Serine, Leucine, Alanine, Arginine, Phenylalanine, Leucine, Glutamic acid, Lysine, Arginine, Lysine, Glycine 325 330 335 Arginine, Isoleucine, Methionine, Serine, Threonine, Alanine, Serine, Proline, Tyrosine, Asparagine, Leucine, Serine, Lysine, Lysine, Serine, Leucine 340 345 350 Aspartic acid, Tyrosine, Alanine, Threonine, Threonine, Methionine, Glutamic acid, Serine, Asparagine, Alanine 355 360 <210> 20 <211> 140 <212> PRT <213> Artificial sequence <400> 20 Met Ser His Asn Ile Asp Leu Phe Arg Lys Tyr Leu Leu Ser Lys Ser 1 5 10 15 Glu Asn His Thr Val Gly Lys Thr Asn Lys Glu Glu Ser Ala Val Met 20 25 30 Lys His Arg Ser Ser Pro Pro Leu Thr Pro Leu Ile Met Pro Pro Lys 35 40 45 Phe Ala Ser Ser Ile Ser Arg Pro Leu Ser Leu Leu Glu Arg Arg Leu 50 55 60 Leu Gln Pro Thr Arg Tyr Leu Gln Lys Ser Ser Thr Thr Gln Leu Thr 65 70 75 80 Ile Phe Tyr Ala Gly Val Val Asn Val Tyr Asp Asn Val Pro Thr Asp 85 90 95 Lys Ala Gln Ala Ile Met Leu Leu Ala Gly Glu Ser Cys Leu Thr Lys 100 105 110 Pro Thr Ala Lys Glu Lys Thr Leu Val Glu Ala Lys Ala Ser Pro Asn 115 120 125 Gln Gln Gln Ala Val Gly Asn Asn Phe Asp Arg His 130 135 140 <210> 21 <211> 194 <212> PRT <213> Artificial sequence <400> 21 Met Ser Arg Ala Ser Val Glu Leu Asp Phe Phe Gly Met Glu Glu Gln 1 5 10 15 Ile Cys Cys Lys Ser Arg Phe Gln Lys Ser Leu Asp Arg Arg Leu Ser 20 25 30 Phe Arg Gly Leu Gln Gly Ala Leu Ser Lys Val Asn Pro Glu Leu Ile 35 40 45 Lys Ser Val Ile Ala Ser Gly Leu Lys Asn Pro Gln Gly Gln Asp Asn 50 55 60 Val Tyr Gln Met Asp Ser Asn Lys Ser Phe Ser Val Pro Ser Ser Pro 65 70 75 80 Lys Glu Thr Gln Ser Leu Phe Pro Ala Leu Pro Leu Leu Thr Pro Ala 85 90 95 Ala Arg Ala Thr Ser Glu Asn Gly Pro Glu Thr Ala Pro Leu Thr Ile 100 105 110 Phe Tyr Asn Gly Thr Val Ser Val Phe Asn Val Pro Arg Asp Lys Ala 115 120 125 Glu Ser Ile Leu Lys Leu Ala Val Glu Val Glu Gly Asn Ser Lys Asn 130 135 140 Val Glu Pro Ile Asp Ser Lys Val Ala Ser Pro Pro Ser Asp Arg Gln 145 150 155 160 Gln Leu Leu Glu Thr Leu Asn Gly Asp Leu Pro Ile Ala Arg Arg Lys 165 170 175 Ser Leu Gln Arg Phe Leu Glu Lys Arg Lys Glu Arg Tyr Phe Leu Phe 180 185 190 Cys Phe <210> 22 <211> 240 <212> PRT <213> Artificial sequence <400> 22 Met Ser Ser Cys Ser Glu Ser Thr Ala Met Lys Pro Ala Arg Ser Leu 1 5 10 15 Glu Lys Pro Ser Phe Ala Gln Thr Cys Asn Leu Leu Ser Gln Tyr Leu 20 25 30 Lys Glu Lys Gly Ser Phe Gly Gly Leu Ser Leu Gly Met Thr Cys Asn 35 40 45 Val Glu Ala Asn Gly Thr Pro Val Val Pro Pro Pro Thr Met Asn Leu 50 55 60 Phe Pro Val Asn Asp Lys Ser Asp Asn Val Cys Gly Arg Asn Gly Gly 65 70 75 80 Asn Pro Lys Asn Leu Ala Ser Met Asp Leu Phe Pro Gln Gln Ala Gly 85 90 95 Leu Ala Ser Lys Asp Asp Ser Pro Asn Lys Leu Glu Pro Gln Thr Ala 100 105 110 Pro Met Thr Ile Phe Tyr Gly Gly Arg Val Ile Val Phe Asn Asp Phe 115 120 125 Pro Ala Asn Lys Ala Lys Glu Ile Met Leu Leu Ala Ser Asn Ser Ser 130 135 140 Ser Gln Ser Asn Asp Ser Phe Asn Pro Ile Pro Phe Thr Ser Ser Ile 145 150 155 160 Ala Arg Ser Pro Ile Lys Ser Ser Ile Gly Val Pro Pro Thr Ser Lys 165 170 175 Pro Val His Pro Ala Gln Arg Ala Val Pro Gly Asp Leu Pro Ile Ala 180 185 190 Arg Arg Ala Ser Leu His Arg Phe Leu Glu Lys Arg Lys Asp Arg Ile 195 200 205 Thr Ala Lys Ala Pro Tyr Gln Ile Ser His Ser Ala Ala Ala Pro Ser 210 215 220 Leu Ser Gly Asp Asn Lys Ser Trp Leu Gly Leu Ala Ala Gln Ser Pro 225 230 235 240 <210> 23 <211> 226 <212> PRT <213> Artificial sequence <400> 23 Met Glu Gly Glu Ala Gly Ser Tyr Glu Asp Val Lys Pro Asn Val Val 1 5 10 15 Ala Lys Gln Ser Asn Gly Asp Val Val Gly Asp Asn Gly Val Gly Asn 20 25 30 Leu Gly Ser Val Glu Ala Pro Asp Phe Leu Ser Lys Lys Asn Phe Gln 35 40 45 Asn Cys Ser Val Ala Thr Pro Ala Ser Gly Leu Asn Ser Thr Gly Pro 50 55 60 Ala Pro Ser Gln Leu Thr Ile Phe Tyr Asp Gly His Val Cys Val Phe 65 70 75 80 Asp Ala Ile Pro Val Glu Lys Val Arg Glu Ile Met Leu Ile Ala Ala 85 90 95 Thr Gly Ala Ala Asn Ser Val Asp Met Lys Lys Val Ala Thr Asp Cys 100 105 110 Ala Thr Thr Ser Pro Val Leu Thr Arg Ser Pro Ser Leu Gln Ser Thr 115 120 125 Ala Thr Ala Thr Ala Leu Ala Ser Pro Gln Ala Gln Val Tyr Pro Thr 130 135 140 Asn Arg Thr Pro Phe Cys Lys Leu Lys Glu Leu Pro Ile Ala Arg Arg 145 150 155 160 His Ser Leu Gln Arg Phe Phe Glu Lys Arg Arg Asp Arg Leu Val Asn 165 170 175 Arg Asn Pro Tyr Pro Asn Pro Ser Thr Pro Lys Ser Phe Asp Asp Thr 180 185 190 Lys Ala Asn Leu Ser Ala Ala Thr Ser Ser Glu Ser Gly Cys Phe Gly 195 200 205 Lys Ser Pro Val Ala Gln Glu Glu Phe His Pro Lys Ala Pro Ala His 210 215 220 Val Ala 225 <210> 24 <211> 226 <212> PRT <213> Artificial sequence <400> 24 Met Thr Lys Asn Phe Leu Pro Asn Met Asp Asn Ser Tyr Glu Ser Leu 1 5 10 15 Lys Pro Asn Leu Gly Ala Ser Thr Ser Lys Ser Asn Ala Lys Pro Cys 20 25 30 Gly Phe Phe Pro Glu Ile Arg Ser Phe Gly Ala Ser Ser Ser Lys Glu 35 40 45 Asp Thr Asn Met Met Thr Asp Phe Arg Lys Pro Ala Lys Val Glu Pro 50 55 60 Lys Asn Ser Gln Met Thr Ile Phe Phe Gly Gly Gln Val Ala Val Phe 65 70 75 80 Asn Asp Phe Pro Ala Asp Lys Phe Lys Glu Ile Met Asp Leu Leu Ala 85 90 95 Ser His Gly Cys Ser Thr Ala Ser Gly Val Val Val Asp Thr Val Met 100 105 110 Glu Lys Val Lys Ser Lys Thr Val Gln Ile Glu Pro Ser Asn His Glu 115 120 125 Ile Pro Asp Leu Asn Val Ser Thr Ala Thr Gly Asn Ser Pro Pro Pro 130 135 140 Pro His Asp Ser Ser Val Glu Trp His Gln Tyr Gly Gly Ser Gly Pro 145 150 155 160 Ser Asp Leu Arg Ile Ala Arg Arg Asn Ser Leu His Lys Phe Phe Glu 165 170 175 Lys Arg Lys Glu Arg Ala Thr Ala Arg Ala Pro Tyr Gln Val Asn Asn 180 185 190 Ala Arg Gly Ser Thr Leu Pro Pro Lys Pro Asp Glu Asn Lys Ser Ser 195 200 205 His Glu Glu Gly Gln Ser Ser Lys Glu Ala Ser Arg Asp Leu Asp Leu 210 215 220 Lys Leu 225 <210> 25 <211> 365 <212> PRT <213> Artificial sequence <400> 25 Met Glu Arg Asp Phe Leu Gly Leu Asn Ser Lys Gln Ser Phe Pro Leu 1 5 10 15 Val Lys Glu Glu Val Glu Glu Ile Gly Phe Thr Lys Ser Leu Gly Ile 20 25 30 Gln Trp Pro Ile Ser Asn Lys Val Ser Ser Ala Val Pro Pro Gln Gln 35 40 45 Met Ser Phe Asp Phe Ala Gln Gly Asp Asn Ala Lys Arg Met Gly Tyr 50 55 60 Asp Ser Ile Val Ser Ser Thr Phe Met His Ile Ser Pro Pro Asp Ala 65 70 75 80 Ala Glu Leu Gln Lys Ser Phe Asn His Asn Arg Gln Arg Val Gly Asn 85 90 95 His Phe Pro Phe Thr Ala Ala Ser Ser Val Gln His Asp Val His His 100 105 110 Val Gln Arg Pro Tyr Asp Met Lys Met Phe Pro Val Ser Asn Gln Ser 115 120 125 Val Pro Val Ser Thr Thr Asn Pro Phe Met Asn Ser His Phe Thr Thr 130 135 140 Thr Ala Met Lys Ser Gln Leu Leu Gly Gly Ile Pro Val Thr Thr Pro 145 150 155 160 Pro His Ser Val Leu Pro Thr Leu Gly Ser Phe Gly Gly Ser Ile Glu 165 170 175 Pro Arg Lys Ser Val Arg Gly Leu Gly Asp Ser Arg Ser Pro Val Gln 180 185 190 Leu Thr Ile Phe Tyr Ala Gly Thr Val Asn Val Tyr Asp Asp Ile Thr 195 200 205 Pro Glu Lys Ala Gln Ala Ile Met Leu Leu Ala Gly Asn Gly Ser Ser 210 215 220 Leu Thr Ser Asn Val Ala His Pro Asn Val Gln Val Gln Ala Pro Ile 225 230 235 240 Ser Lys Pro Leu Gln Phe Glu Thr Leu Pro Thr Asn His Phe Thr Asn 245 250 255 Ala Glu Leu Cys Ser Gly Ile Pro Ser Pro Leu Ser Val Ser Ser His 260 265 270 Thr Gly Val Gln Ser Arg Ser Gly Ser Thr Ser Thr Asp Glu Lys Thr 275 280 285 Val Cys Lys Thr Thr Gly Ser Leu Thr Thr Pro Ile Ser Leu Val Glu 290 295 300 Ser Pro Lys Leu Ala Asn Thr Met Gly Pro Val Thr Pro Thr Ser Met 305 310 315 320 Met Pro Ser Val Pro Gln Ala Arg Lys Ala Ser Leu Ala Trp Phe Leu 325 330 335 Glu Lys Arg Lys Glu Arg Ile Met Thr Ala Ser Pro Tyr Asp Leu Ser 340 345 350 Lys Lys Pro Pro His Cys Thr Thr Gln Gly Ser Asn Ala 355 360 365 <210> 26 <211> 119 <212> PRT <213> Artificial sequence <400> 26 Met Arg Arg Asn Cys Asn Leu Glu Leu Arg Leu Leu Pro Ser Ser Tyr 1 5 10 15 Pro Ser Asp Ser His Asp Met Met Glu Ala Arg Ile Glu Ser Pro Glu 20 25 30 Thr Gln Gln Gln Gln Leu Thr Ile Phe Tyr Asn Gly Arg Val Cys Val 35 40 45 Ser Asp Val Thr Glu Leu Gln Ala Lys Ala Ile Leu Ile Leu Ala Asn 50 55 60 Arg Glu Arg Asp Glu Arg Met Lys Ser Pro Thr Gly Cys Glu Pro Val 65 70 75 80 Ser Pro Thr Leu Lys Ser Gln Val Asn Ser Pro Asn Thr Ser Leu Ser 85 90 95 Met Lys Arg Ser Leu Gln Arg Phe Leu Gln Lys Arg Lys Thr Arg Ile 100 105 110 Gln Ala Thr Ser Pro Tyr His 115 <210> 27 <211> 263 <212> PRT <213> Artificial sequence <400> 27 Met Asn Met Ser Cys Ser Pro Glu Phe Met Val Gln Lys Pro Ala Arg 1 5 10 15 Ser Pro Glu Lys Thr Ser Phe Thr Gln Thr Cys Asn Leu Leu Ser Gln 20 25 30 Tyr Leu Lys Glu Lys Gly Ser Phe Gly Asp Leu Ser Leu Gly Met Thr 35 40 45 Cys Asn Val Glu Ala Asn Glu Thr Pro Glu Met Leu Arg Pro Thr Met 50 55 60 Asn Leu Phe Pro Val Asn Gly Lys Ser Gly Asp Asp Cys His Ala Ala 65 70 75 80 Pro Pro Pro Arg Lys Leu Arg Ser Met Asp Leu Phe Pro Asn Gln Ala 85 90 95 Ala Phe Ser Ser Pro Lys Asp Asp Ala Leu Lys Ser Thr Met Asn Lys 100 105 110 Leu Gly Ser Ser Val Glu Pro Gln Thr Ala Gln Met Thr Ile Phe Tyr 115 120 125 Gly Gly Gln Val Ile Val Phe Asn Asp Phe Pro Ala Asp Lys Ala Lys 130 135 140 Glu Ile Met Leu Leu Ala Gly Lys Gly Ser Ser Gln Asn Asn Ser Phe 145 150 155 160 Asn Pro Asn Pro Pro His Ile Asn Ala Pro Phe Thr Ser Thr Ile Ala 165 170 175 Thr Ser Pro Ile Glu Ser Gly Ile Gly Val Pro Pro Thr Pro Asn Phe 180 185 190 Ser Thr Thr Val Thr Gln Glu Cys Ile Arg Ser Ala Gln Arg Pro Ile 195 200 205 Pro Gly Asp Leu Pro Ile Ala Arg Arg Ala Ser Leu His Arg Phe Leu 210 215 220 Glu Lys Arg Lys Asp Arg Met Thr Thr Ser Ala Pro Tyr Gln Ile Ser 225 230 235 240 Asn Ser Thr Ala Ser Ser Ser Lys Pro Gly Asn Asp Lys Ser Trp Leu 245 250 255 Gly Leu Ala Ala Gln Ser Pro 260 <210> 28 <211> 228 <212> PRT <213> Artificial sequence <400> 28 Met Glu Glu Glu Ala Glu Ser Arg Glu Glu Val Lys Pro Asn Val Val 1 5 10 15 Val Lys Glu Thr Asn Gly Asp Ile Val Gly Asp Asn Asp Ala Gly Lys 20 25 30 Leu Gly Thr Ile Glu Thr Pro Asp Phe Leu Ser Gln Lys Ile Ser His 35 40 45 Asn Cys Ser Leu Pro Ile Leu Ala Ser Gly Val Asn Thr Thr Ser Leu 50 55 60 Ala Gln Ser Gln Leu Thr Ile Phe Tyr Gly Gly Lys Val Ser Val Phe 65 70 75 80 Asp Ala Ile Ser Ala Glu Lys Ile Gln Glu Ile Met Leu Ile Ala Ala 85 90 95 Ala Val Ala Ala Ala Asp Val Gly Ser Val Asp Met Lys Asn Ala Ala 100 105 110 Thr Asp Tyr Ala Thr Ile Ser Pro Ala Leu Thr Arg Cys Pro Ser Leu 115 120 125 Gln Ser Thr Ala Thr Ala Leu Ala Ser Pro Gln Ala Gln Leu Tyr Pro 130 135 140 Phe Pro Arg Thr Ser Phe Ser Lys Leu Gln Ala Glu Leu Pro Ile Ala 145 150 155 160 Arg Arg His Ser Leu Gln Arg Phe Leu Glu Lys Arg Arg Asp Arg Leu 165 170 175 Val Asn Lys Asn Pro Tyr Pro Gly Pro Ser Thr Pro Lys Met Val Asp 180 185 190 Gly Ala Lys Ala Asp Val Ser Ala Thr Thr Ser Pro Glu Ser Gly Cys 195 200 205 Phe Lys Ala Ser Pro Ile Arg Gln Glu Asp Ile Gln Pro Lys Ala Pro 210 215 220 Ala His Val Ala 225 <210> 29 <211> 223 <212> PRT <213> Artificial sequence <400> 29 Met Phe Gly Ser Pro Glu Lys Pro Met Phe Lys Arg Thr Cys Ser Leu 1 5 10 15 Leu Ser Gln Tyr Leu Lys Glu Lys Gly Ser Phe Gly Asp Leu Thr Leu 20 25 30 Gly Ile Thr Cys Ser His Asn Val Glu Lys Gly Met Pro Glu Ile Val 35 40 45 Arg Pro Ala Thr Glu Thr Arg Thr Thr Thr Met Asp Leu Phe Pro Gln 50 55 60 Gly Ala Gly Phe Ser Ala Asp Asn Gly Ser Arg Arg Val Gly Val Asp 65 70 75 80 Val Asp Pro Gln Asn Ala Ala Ala Pro Met Thr Ile Phe Tyr Cys Gly 85 90 95 Gln Val Ile Val Phe Asn Asp Phe Pro Ala Asp Lys Ala Lys Glu Ile 100 105 110 Met Ala Leu Ala Ser Lys Cys Ser Ser Glu Asn Pro Lys Thr Asn Thr 115 120 125 Phe Val Pro Gly Ser Pro Asn Glu Ser Gly Leu Gly Val His Ser Asn 130 135 140 Ser Asn Asp Gln Val Pro Asn Ile Arg Thr Asn Leu Pro Thr Ser Arg 145 150 155 160 Glu Cys Val Arg Pro Ile Pro Gly Asp Leu Pro Ile Ala Arg Arg Ala 165 170 175 Ser Leu His Arg Phe Leu Glu Lys Arg Lys Asp Arg Ile Thr Ser Lys 180 185 190 Ala Pro Tyr Pro Ile Asn Gly Ser Ala Gly Ala Ser Pro Pro Lys Pro 195 200 205 Gly Asn Ser Lys Pro Trp Leu Gly Leu Ala Val Glu Ser Leu Gln 210 215 220 <210> 30 <211> 125 <212> PRT <213> Artificial sequence <400> 30 Met Arg Arg Lys Cys Asn Leu Glu Leu Glu Leu Phe Pro Ser Arg Ile 1 5 10 15 Phe Pro Gly His Arg Gln Asn Met Val Glu Glu Ser Lys Arg Ile Pro 20 25 30 Gln Asn Gln Gln Gln Gln Leu Thr Ile Phe Tyr Asn Gly Arg Val Cys 35 40 45 Val Cys Asp Val Thr Glu Leu Gln Ala Arg Ala Ile Leu Met Arg Ala 50 55 60 Asn Arg Glu Ala Asp Glu Arg Ile Lys Thr Pro Thr Gly Ser Glu Pro 65 70 75 80 Asp Ser Pro Thr Ser Ser Ser Ser Thr Ser Pro Ser Arg Leu Cys Ser 85 90 95 Pro Asn Ala Gly Leu Ser Met Lys Lys Ser Leu Gln Arg Phe Leu Gln 100 105 110 Lys Arg Lys Asn Arg Ile Gln Ala Thr Ser Pro Tyr His 115 120 125 <210> 31 <211> 120 <212> PRT <213> Artificial sequence <400> 31 Met Arg Arg Asn Cys Asn Leu Glu Leu Arg Leu His Pro Ser Ser Tyr 1 5 10 15 Ser Gly Gly Asp Arg Val Glu Glu Ser Ser Glu Asn Pro Glu Asn Gln 20 25 30 Gln Gln Gln Leu Thr Ile Phe Tyr Asn Gly Arg Val Cys Val Cys Asp 35 40 45 Val Thr Glu Ile Gln Ala Arg Ala Ile Leu Met Ile Ala Asn Arg Glu 50 55 60 Thr Asp Glu Arg Leu Arg Thr Pro Arg Ser Gly Ser Gly Pro Ala Ser 65 70 75 80 Pro Thr Val His Ser Gln Ile Asn Ser Pro Asn Asn Gly Leu Ser Thr 85 90 95 Ser Met Lys Arg Ser Leu Gln Arg Phe Leu Gln Lys Arg Lys Asn Arg 100 105 110 Ile Gln Ala Thr Ser Pro Tyr His 115 120 <210> 32 <211> 270 <212> PRT <213> Artificial sequence <400> 32 Met Ser Asn Leu Gly Gln Lys Ser Pro Asp Arg Ala Ser Phe Val Asn 1 5 10 15 Leu Leu Ser Gln Tyr Leu Lys Glu Lys Arg Asn Leu Gly Asp Phe Ser 20 25 30 Leu Gly Met Thr Ser Lys Pro Asp Ala Lys Gly Leu Glu Thr Ser Arg 35 40 45 Gln His Ala Lys Asn Met Asn Phe Leu Ser Asn Met Pro Asn Cys Ser 50 55 60 Glu Ser Ser Arg Pro Asn Leu Val Ala Ser Thr Ser Asn Val Lys Ser 65 70 75 80 Ser Asp Phe Phe Pro Glu Ile Gly Ser Phe Gly Ala Ser Ser Ser Lys 85 90 95 Glu Asp Thr Phe Asn Lys Thr Asp Phe Met Lys Ser Ala Ala Val Glu 100 105 110 Pro Lys Asn Ala Gln Leu Thr Ile Phe Phe Gly Gly Gln Val Phe Val 115 120 125 Tyr Asn Asp Phe Pro Ala Asp Lys Val Lys Glu Ile Met Ala Val Ala 130 135 140 Asn Arg Gly Trp Ser Thr Ser Cys Ser Gly Val Val Ala Asp Ser Ala 145 150 155 160 Met Glu Lys Leu Asn Ala Asn Leu Asp Lys Ile Asp Tyr Ser Ser Pro 165 170 175 His Ile Pro Asp Leu Asn Ile Thr Ser Ala Thr Ala Asn Ser Pro Ala 180 185 190 Gln Asp Pro Ser Val Glu Arg Cys Gln Tyr Val Gly Ser Asp Leu Arg 195 200 205 Ile Ala Arg Arg Asn Ser Leu His Lys Phe Phe Glu Lys Arg Lys Asp 210 215 220 Arg Ala Ala Met Arg Ala Pro Tyr Gln Leu Asn Asn His Gln Gly Ser 225 230 235 240 Pro Pro Pro Pro Lys Pro Asp Glu Asn Lys Pro Ser His Glu Glu Gly 245 250 255 Gln Ser Ser Lys Glu Thr Pro Arg Asp Ile Asp Leu Asn Leu 260 265 270 <210> 33 <211> 365 <212> PRT <213> Artificial sequence <400> 33 Met Glu Ala Gly Val Thr Thr Thr Ala Thr Thr Thr Ala Ser Phe Ser 1 5 10 15 Ser Ile Leu Asp Lys Pro Leu Ser Gln Leu Thr Glu Glu Asp Ile Ser 20 25 30 Gln Leu Thr Arg Glu Asp Cys Arg Lys Phe Leu Lys Glu Lys Gly Met 35 40 45 Arg Arg Pro Ser Trp Asn Lys Ser Gln Ala Ile Gln Gln Val Ile Ser 50 55 60 Phe Lys Ala Leu Leu Glu Ser Asn Glu Asp Ser Gly Ala Gly Ala Arg 65 70 75 80 Arg Lys Ile Leu Val Cys Pro Pro Pro Ser His Phe Pro Pro Gln Asn 85 90 95 Ala Val Ala Ser Asn Ser Gly Glu Ser Val Lys Glu Ala Val Phe Gly 100 105 110 Glu Glu Glu Ser Leu Tyr Gly Gln Lys Asp Leu Ser Leu Lys Ala Ala 115 120 125 Pro Val Val Gln Met Asn Cys Gln Gly Gly Asp Thr Asp Asp Lys Thr 130 135 140 Leucine, Serine, Proline, Serine, Leucine, Glycine, Serine, Proline, Arginine, Glutamic acid, Tyrosine, Serine, Lysine, Leucine, Proline, Glycine 145 150 155 160 Arginine, Serine, Glutamine, Cysteine, Glutamic acid, Threonine, Asparagine, Glutamic acid, Leucine, Glycine, Glycine, Glutamine, Methionine, Threonine, Isoleucine, Phenylalanine 165 170 175 Tyrosine, Cysteine, Glycine, Lysine, Isoleucine, Asparagine, Valine, Tyrosine, Aspartic acid, Glycine, Valine, Proline, Leucine, Alanine, Lysine, Alanine 180 185 190 Arginine, Alanine, Isoleucine, Methionine, Histidine, Leucine, Alanine, Alanine, Serine, Proline, Isoleucine, Aspartic acid, Phenylalanine, Proline, Glutamine, Glycine 195 200 205 Asparagine, Leucine, Cysteine, Asparagine, Glutamine, Asparagine, Glycine, Alanine, Phenylalanine, Arginine, Serine, Phenylalanine, Leucine, Glycine, Histidine, Valine 210 215 220 Glutamine, Glutamic acid, Alanine, Glutamic acid, Aspartic acid, Lysine, Asparagine, Aspartic acid, Leucine, Threonine, Serine, Serine, Isoleucine, Alanine, Leucine, Asparagine 225 230 235 240 Leucine, Asparagine, Serine, Histidine, Isoleucine, Methionine, Histidine, Threonine, Glutamic acid, Lysine, Methionine, Threonine, Glutamic acid, Tyrosine, Glutamine, Glutamine 245 250 255 Glutamine, Phenylalanine, Arginine, Glycine, Lysine, Alanine, Asparagine, Isoleucine, Serine, Arginine, Aspartic acid, Serine, Aspartic acid, Valine, Aspartic acid, Glycine 260 265 270 Glutamine, Valine, Serine, Arginine, Lysine, Valine, Serine, Leucine, Glutamine, Arginine, Tyrosine, Leucine, Glutamic acid, Lysine, Arginine, Lysine 275 280 285 Aspartic acid, Arginine, Glycine, Arginine, Phenylalanine, Phenylalanine, Lysine, Glycine, Arginine, Lysine, Asparagine, Alanine, Glycine, Glutamine, Threonine, Leucine 290 295 300 Ser Ser Ser Glu Met Tyr Leu Asn His Gln Ile Arg Ala His Tyr Leu 305 310 315 320 Asn Gly Gln Thr Asn Gln Ser Arg Thr Ser Ser Pro Pro Gln Ser Gly 325 330 335 Val Pro His Ala Phe Tyr Ser Ser Ala Asp Asn Gln Glu Leu Val Asn 340 345 350 Phe Ser Val Asp Leu Asn Asp Glu Gly Gly Gln Glu His 355 360 365 <210> 34 <211> 337 <212> PRT <213> Artificial sequence <400> 34 Met Ser Pro Gly Glu Thr Val Ser Arg Ser Pro Leu Asp Lys Pro Leu 1 5 10 15 Asn Gln Leu Thr Glu Asp Asp Ile Ser Gln Val Thr Arg Glu Asp Cys 20 25 30 Arg Arg Tyr Leu Lys Glu Lys Gly Met Arg Arg Pro Ser Trp Asn Lys 35 40 45 Ser Gln Ala Ile Gln Gln Val Ile Ser Leu Lys Thr Leu Leu Glu Thr 50 55 60 Thr Ser Asp Ser Asp Ala Val Glu Ala Arg Lys Lys Leu Tyr Pro Pro 65 70 75 80 Cys Pro Glu Tyr Pro Pro Arg Val Val Ser Asp Ser Asn Val Leu Pro 85 90 95 Arg Glu Met Thr Pro Asn Asn Gly Ile Leu Val Pro Val Ser Glu Ser 100 105 110 Val Pro Cys Pro Leu Ser Asn Pro Ser Lys Ser Asp Phe Ser Gly Asp 115 120 125 Asn Ser Gly Arg Thr Val Ile Ser Gly Asn Asp Ser Val Ser Pro Arg 130 135 140 Ile Ala Gly Ala Ala Lys Glu Pro Ala Gly Gln Met Thr Ile Phe Tyr 145 150 155 160 Cys Gly Lys Val Asn Val Tyr Asp Asp Met Pro Gly Cys Lys Ala Glu 165 170 175 Ala Ile Met Gln Leu Ala Ala Ser Pro Val Ser Phe Pro His Glu Ile 180 185 190 Leu Ala Asp Gln Arg Ser Thr Pro Trp Ser Ile Pro Cys His Ser Gln 195 200 205 Ala Ala Ser Val Lys Thr Thr Pro Cys Ser Gln Met Val Ile Leu Pro 210 215 220 Pro Gln Gln Thr Glu Asn Cys Gln Phe Pro Arg Glu Glu Ser Asn Ala 225 230 235 240 Ser Leu Glu Asp Ser Leu Glu Gly Pro Thr Ser Arg Lys Ala Leu Val 245 250 255 His Arg Tyr Leu Glu Lys Lys Lys Asp Arg Phe Lys Asn Lys Arg Lys 260 265 270 Leu Ala Met Ser Ser Ser Pro Thr Leu Asp Ile Tyr Leu Asn Gln Val 275 280 285 Gly Asp Gln Phe Ser Asn Glu Gln Leu Lys Gln Ser Glu Pro Tyr Tyr 290 295 300 Ser Pro Gln Ala Glu Ala His Arg Met Pro Leu Glu Cys Ser Ser Ile 305 310 315 320 Glu Asn Val Ala Lys Ile Pro Arg Leu Thr Thr Asp Gly Lys Gly Asn 325 330 335 Arg <210> 35 <211> 371 <212> PRT <213> Artificial sequence <400> 35 Met Glu Arg Asp Phe Leu Gly Leu Arg Ser Lys Asn Ala Ala Ile Thr 1 5 10 15 Ile Lys Val Glu Pro Ser Asp Ala Gln Pro Asn Asp Ser Val Trp Leu 20 25 30 Arg Gly Ser Gly Met Gln Leu Ser Phe Thr Asn Lys Val Ser Thr Val 35 40 45 Pro Gln Phe Leu Ser Phe Asp Gly Ala Arg Asp Asp Lys Pro Arg Lys 50 55 60 Ala Thr His Asp Thr Leu Leu Ser Ser Gly Phe Met Thr Ile Pro Thr 65 70 75 80 Thr Asp Ser Asn Lys Lys Pro His Pro Gly Leu Thr Gln Lys Gln Gly 85 90 95 Gly Asn His Tyr Ala Ala Thr Thr Tyr Gly Leu Gln Gln Leu Asp Val 100 105 110 His Gln Ser Arg Leu Ser His Glu Ala Arg Ile Phe Pro Ser Ser Ser 115 120 125 Gln Leu Asn His Thr Ile Thr Val Ser Met Asn Thr Pro Leu Leu Gln 130 135 140 Pro His Leu Ala Ser Pro Gly Gln Asn Ile Ile Gly His Thr Ile Asn 145 150 155 160 Pro Gln Pro Phe Thr Gly Val Pro Ile Met Ala Ala Pro Val Ser Val 165 170 175 Val Pro Pro Ser Ser Pro Ile Ile Gly Thr Thr Asp Leu Arg Asn Ala 180 185 190 Ala Lys Ser Ser Lys Ala Pro Ala Gln Leu Thr Ile Phe Tyr Ala Gly 195 200 205 Ser Val Cys Val Tyr Asp Asp Val Ser Pro Asp Lys Ala Gln Ala Ile 210 215 220 Met Leu Leu Ala Gly Asn Gly Ser Ser Ala Thr Gln Ser Lys Thr Ala 225 230 235 240 Pro Val Thr Gln Arg Gln Thr His Ile Pro Arg Pro Cys Thr Leu Ser 245 250 255 Pro Phe Ser Gly Leu Pro Asn His Leu Ser Ala Thr Ser His Val Ser 260 265 270 Leu Gln Pro Val Ala Gly Ser Ser Gly Thr Asn Glu Leu Thr Ala Ala 275 280 285 Thr Arg Ile Gly Ala Leu Ala Ser Ala Asn Asn Gln Pro Asp Pro Pro 290 295 300 Lys Leu Val Asn Pro Ala Val Ala Val Pro Gln Ala Arg Lys Ala Ser 305 310 315 320 Leu Ala Arg Phe Leu Glu Lys Arg Lys Glu Arg Val Ser Asn Thr Ser 325 330 335 Pro Tyr Asn Ile Cys Lys Arg Ser Pro Glu Ser Gly Pro Leu Ala Ser 340 345 350 Asp Gly Ile Ser Phe Pro Val Thr Ser Ala Gly Ser Ser Pro Leu Gln 355 360 365 Ala Ile Asn 370 <210> 36 <211> 630 <212> DNA <213> Artificial sequence <400> 36 atgtccagag ctaccgtcga gcttgatttc ttcggcatgg agaaagccaa ttcctgcaaa 60 tctcagttcc agaaattcct tgatcgccgc cggagcttcc gaggtattca aggcgccatt 120 tcgaagatga atccagagct tatcaaatct gtgattgctt ccggttcgac gacccggaat 180 ccggttgatt ggagaaaatc cttttcggtt ccgtcgagtc ctaaagaaga tcggagtact 240 tcgcttcctt ctttacctct actcaatcct gctttgaggt ctattccttc tgaggagagt 300 cctgaaacag ctccgttgac gattttttac aacggaacgg tttccgtaat caatgtacct 360 cgagacaagg ttcgcggaag catcttcaaa cttgccgttg aaggaagctc aaaaaacatt 420 gaatcagtag actcatcaaa agctgcaaat ccttcaagtg atcaacaaaa cctcctcgaa 480 gctcgtaatg gagatttgcc aattgctaga agaaagtcgc tgcaaagatt tctggaaaag 540 cgcaaagaga ggttagatat attactgtgt tggatccgaa tttacagaat ttttactaat 600 tttctaaaaa aaaaaaaaaa caacaactga 630 <210> 37 <211> 1092 <212> DNA <213> Artificial sequence <400> 37 atggagagag attttctggg tttgaattca aatgaaccat tggctgtggt taaggatgat 60 gttaacactg ataagtacaa acaaataggt tttactaaaa gctcagggat acaatggccc 120 ttttcaaaca aagttttcgc tgttccacag ctgatgaact ttaattttgc ccaaggggat 180 aaaaccaaaa agactggata tgattcaaaa gtatcccctc tcttcatgcc tatatcaaca 240 atggatgctt ctgaacttca gaaatcgttc aatcacaatt ggaatggtgg gggtcatttt 300 tcattgactg attctcatgt tcaacacaac acgaacatgt ttccggcatc aaaccagaca 360 atttctgttt ctgggagcaa cccgttcgtg aagaatcatt tcactaccac tggtcaaaac 420 tttcctgcta atactataaa accacaattt cttggagggg ttccagtgac aactccacat 480 tcagttcttc ccacccttgg cactgttggt ggaagtgttg aaccatgcgc ccagacctct 540 ggatcacctg ctcaattgac tatcttttat gcgggtgaag tgaatgtctt tgatgatatt 600 acccctgaga aggctcaagc tatcatgttt ttggctggga acgggtcttc catggcttct 660 aattcagctt atccaaaacc cccggtccag acacctattt tgaaaccagt tcaagttgat 720 agtgttcctg cgaaccagct cataaataca caactgagct ttggtatgcc aagcccttta 780 cctgtttcat ctcatgctgg tatgcagtct tggagtggat ccaccagtac cgaagaacag 840 ataatatgca aggcctcagt acctccaact ccatctaccc ctattagcaa attggagagt 900 ccaaatttgg taaataccat gggatctgat gctgccaccg gcatgatgcc ttctgttcca 960 caggctcgca aagcttcctt ggctcggttt ttggagaagc gcaaggggag gataatgagt 1020 acagcatcac catacaatct tattagcaag aagtctctag attatgcaac taccatggaa 1080 tcgaatgcat aa 1092 <210> 38 <211> 594 <212> DNA <213> Artificial sequence <400> 38 atgtcgagag ctagcgtcga gcttgatttc tttggaatgg agaaacagaa cttctgcaaa 60 tctcggtttc agaaatctct cgatcgccgt ctaagctttc gaggtcttca aggcgccctt 120 tcgaaggtga atcctgagct tattaaaacc gtgattgcct ctagtttgaa gaacccgcag 180 ggtcaggata atgtgtatca gatggactcc aacaagtctt tttcggtccc ttccagtcct 240 aaagaaactc agagtctgtt tccagcttta cctcttctta accctgctgc cagggctaca 300 tccgagaatg gtcctgaaat agctccctta accattttct acaacggaac ggtttccgtt 360 tttaatgtac ctcgagataa ggcggagagc atcttaaaac tcgcggttga agttgaagga 420 aactcaaaaa atgttgaacc aatagactca aaagtggcaa gtcctcctag cgatcgacaa 480 cagcttcttg aaactctcaa tggagatttg ccaatcgccc ggagaaagtc tttgcagaga 540 tttctggaaa agcgcaaaga gaggatgact tgtgcatctc catatgcttg ctag 594 <210> 39 <211> 723 <212> DNA <213> Artificial sequence <400> 39 atgtcgtctt gctcggaatc tacggctatg aaaccggcta ggtcaccgga gaagccaagt 60 Met - Ser - Leu - Ala - Arg - Ile - Tyr - Gly - Tyr - Glu - Thr - Gly - Leu - Val - Thr - Gly - Glu - Ala - Lys tttgcccaaa cttgtaattt gttgagccag tacttgaagg agaaaggtag ctttggggat 120 Phe - Ala - Gln - Leu - Val - Asn - Phe - Leu - Glu - Pro - Thr - Leu - Lys - Glu - Lys - Gly - Ser - Leu - Gly - Asp ctatctctgg gaatgacatg caacgttgaa gctaatggga caccagtggt gccgcctccg 180 Leu - Ile - Leu - Gly - Asn - Asp - Met - Gln - Arg - Glu - Ala - Asn - Gly - Thr - Gln - Gly - Ala - Ala - Pro - Ser accatgaatc tgttccctct caatgacaag tccgatgatg tttgtggccg aaacggtggg 240 Thr - Met - Asn - Ser - Val - Pro - Ser - Gln - Asp - Gln - Ser - Asp - Met - Phe - Val - Gly - Glu - Thr - Gly - Gly aatcctaaga acttgacatc catggatttg ttcccccaac aagctggttt ggcttctaaa 300 Asn - Pro - Lys - Asn - Leu - Asp - Ile - His - Gly - Phe - Val - Pro - Asn - Lys - Leu - Val - Trp - Ala - Ser - Lys gatgatagtc ccaacaagtt ggagcctcag actgcaccaa tgaccatctt ttatggtgga 360 Asp - Asp - Ser - Pro - Asn - Lys - Leu - Glu - Pro - Gln - Thr - Ala - Gln - Met - Thr - Ile - Phe - Met - Gly - Glu caagtgattg tgttcaatga ttttccagca aacaaagcta aggaaatcat gcttttagct 420 Gln - Val - Ile - Val - Phe - Asn - Asp - Phe - Pro - Gln - Asn - Lys - Ala - Lys - Glu - Ile - Met - Leu - Ala agcaacagca gctcacaaag caacaacagc ttcaacccca tcccttttac ttctagcata 480 Ser - Asn - Ser - Ala - His - Lys - Ala - Asn - Asn - Ser - Phe - Asn - Pro - Ile - Pro - Phe - Thr - Phe - Ser - Ile gccagaagtc caatcgaatc aagcattgga gttcctccta cctcaaaacc ggttcaccct 540 Ala - Glu - Val - Pro - Asn - Arg - Ile - Lys - His - Gly - Val - Pro - Pro - Thr - Gln - Thr - Gly - Ser - Thr - Pro gctcagcgag ctgtccctgg cgatctgcca attgcaagga gagcttcact gcatcggttc 600 Ala - Gln - Glu - Leu - Val - Pro - Gly - Asp - Leu - Pro - Ile - Ala - Lys - Glu - Leu - His - Ala - Arg - Phe ctagagaagc gaaaggatag gatcactgca aaagcgccat accagataaa taactcagct 660 Leu - Arg - Glu - Arg - Lys - Asp - Arg - Asp - His - Ala - Lys - Ala - His - Thr - Gln - Asp - Asn - Thr - Ser - Ala gcagctcctt ccctgtccgg cgataacaag tcatggctcg gtttggctgc tcaatcacca 720 Ala - Ala - Pro - Pro - Cys - Pro - Gly - Asp - Asn - Lys - Ser - Met - Ala - Arg - Phe - Gly - Cys - Ser - Asn - Thr tag 723 Stop <210> 40 <211> 651 <212> DNA <213> Artificial sequence <400> 40 atggaagggg aagctggttc atacgaggac gttaagccaa acgtcgtcgt taaacaaagt 60 aatggcgatg ttgtcgggga taatggtgtt ggaaatctgg gatccgttga cgcccccgat 120 tttctttcca agaaaaattt ccagaactgc cctgctccat ctcaactaac catcttctat 180 gatggacacg tttgtgtgtt tgacgcaatt cctgtggaaa aggtgcggga gattatgctt 240 attgccgcaa ctgctgctgc tggtgctgct aactctgttg acatgaagaa agttgcaact 300 gattgtgcca ccacctcacc tgttcttacg aggtctcctt cgcttcaaag tactgctact 360 gcaactgctc tggcttcacc acaggcacag gtgtacccta ttaacaggac ccctttctgc 420 aaactgaaag aactaccaat tgcaaggaga cactcccttc agcgattttt tgagaagcgg 480 cgggacaggc tggtgaacag gaatccatat cctaatccat caacaccaaa atcatttgat 540 gacaccaaag ctaacctcag tgctgcaact tcaccagaat cgggttgctt tggtaaatcg 600 cctgttgctc aagaagaatt ccatccaaaa gctccagctc atgttgcata a 651 <210> 41 <211> 681 <212> DNA <213> Artificial sequence <400> 41 atgacgaaga actttttgcc caacatggaa aattcttatg agagtttaaa accgaatctc 60 ggggcatcta catcgaaatc aaatgcgaaa cgttgtggtt ttttcccaga gattaggagt 120 tttggtgtct ccagttccaa ggaagatacc aacaggatga cagatttcag gaaaccagca 180 aaagtggaac caaagaattc ccagatgacc atattttttg gcagtcaagt ggcagtattt 240 aatgacttcc cagctgacaa gttcaaggaa atcatggact tattacctag ccatggatgc 300 tcaaccgcaa gcggtggtgt tgttgatact gtcatggaaa aagttaagtc taaaatagtc 360 caaatcgagc ccagcaatca tgaaattcca gacctgaatg ttgctactgc gactgggaat 420 agtcctcctc cccctcatga ttcttccgtt gagtggcatc aatatggcgg ttcaggatct 480 tcagatcttc gaattgcaag aagaaattca cttcacaagt ttttcgaaaa gagaaaagaa 540 agggcaacag caagagcgcc ataccaagta aacaacgcaa gaggatcgac tccaccacct 600 aaacccaatg aaaacaaatc atctcacgag gaaggtcaat catcgaaaga agcctcaaga 660 gatcttgatc tcaagttata g 681 <210> 42 <211> 360 <212> DNA <213> Artificial sequence <400> 42 atgagacgaa actgcaactt ggaacttcgt cttcttcctt ctagttatcc cagcgacagt 60 cacgacatga tggaagaaag aatcgaaagc ccaaaaaccc agcagcaaca gctaacaatt 120 ttctacaatg gaagagtttg cgtatctgat gttacagagc ttcaggcaaa agccattcta 180 atgttagcaa accgggaaag agatgaaaga atgaaaagtc caacgggatg ggaaccggtg 240 tcgccgacgt taaaatctca ggtgaaatgc ccaaacacag cgctttccat gaagagatcg 300 cttcaacggt tcctccagaa acgaaagacg cgaatccaag ctacctctcc ttaccattaa 360 <210> 43 <211> 1095 <212> DNA <213> Artificial sequence <400> 43 atggagagag attttctggg tttgaactca aaacaatcct tccctttggt taaagaagaa 60 gttgaagaaa ttggttttac caaaagcttg gggattcaat ggccaatttc aaacaaggtt 120 tcttcagctg ttccaccaca gcagatgagt tttgattttg ctcaagtaga taacgctaaa 180 agaattggat atgattcaat agtatcccct gctttcatgc atatctcacc accagatgct 240 gctcaacttc agaaatcctt caatcacaac aggcaacgag taggcaatca tttcccattc 300 acagctgctt ctgttcagca tgatgcacat catgttcaac ggccttatga catgaaaatg 360 tttccggtct cgaaccaatc agttcccgtt tcaaccacca acccatttat gaataatcat 420 ttcactacta ctgctatgaa atcacaattg cttggaggaa ttccagttac gagtccgccg 480 cattcggttc ttcccaccct cagttccttt ggtggaagca tcgaaccacg gaaaagtgtc 540 aggggccttg gggactccag atcacctgtt caattgacta tcttttatgc tggtaccgtg 600 aatgtttacg atgatatcac ccccgagaag gctcaggcta tcatgttact tgctgggaat 660 ggatcttcct tgacttctaa tgtggcacat ccaaaagtcc aagtccaggc accaatttca 720 aaaccacttc aattcgaaaa tcttcccacg aatcatttca cgaatgaaca actttgttct 780 ggtatcccaa gccctttatc ggtttcatct cacactggtg ttcaatccag gagtggatcc 840 actagtactg atgaaaagac cgtatgcaag accacaggaa gtctgacaac ccctattagc 900 ctagtggagt ctcctaaatt ggcaaatacc atgggacctg ttaccccaac cagcattatg 960 cctactgtgc cacaggctcg caaagcatcc ttggctcggt ttttggagaa gcgcaaggag 1020 aggattatga ctgcatcacc atatgatcta agcaagaaac ctccacattg tacaacccag 1080 ggatcaaatg catga 1095 <210> 44 <211> 792 <212> DNA <213> Artificial sequence <400> 44 atgaatatgt cgtgttcacc ggaatttttg gttcagaaac cgacgaggtc accggagaag 60 acaagcttta cccaaacttg taacttgttg agtcagtact tgaaggagaa aggtagcttt 120 ggagatctaa gtctgggtat gacatgcaac gatgaagcta atgagactcc cgagatgctg 180 cgtcccacca tgaatttgtt tcctgtcaat gagaaatccg gtgatgattg ccttgcggcg 240 cctcctcctc ggaaactgag atccatggat ttgttcccta atcaagctgc tttttcatca 300 cctaaagatg atgctctgaa aagcaccatg aacaaattgg gttcttcagt ggagcctcaa 360 actgcacaga tgaccatctt ttatggtgga caagtgattg tgttcaacga tttcccggcc 420 gacaaagcta aggagatcat gcttttagct ggcaaaggca gctctcaaag caatagcttc 480 aacaccaatc ctccccacat caatgccccc tttacttcta ccatagcaac aagtccaatt 540 gagtccggta tcggggttcc tcctacccct aatttcagca ccacggtaac tcaagaatgc 600 atacgatctg ctcagcgatc cattcccggc gatctaccaa ttgcaaggag agcttcactc 660 caccggttcc ttgagaagag aaaggatagg atgaccacga gggcaccata cgagataagt 720 aactccacag catcttcatc caagccaggc gacgacaagt catggctcgg tttggctgct 780 caatctccgt ag 792 <210> 45 <211> 687 <212> DNA <213> Artificial sequence <400> 45 atggaagaag aggctgaatc acgcgaggaa gtcaagccaa acgtcgtcgt taaagagaca 60 aatggagata ttgtcggcga taacgatgca ggaaaattgg gtaccattga aacccctgat 120 tttctttccc agaaaatatt ccataactgt tccctaccaa ttctggcatc tggagtaatt 180 accacaagcc ctgctcaatc ccaactcacc atcttctatg gcggaaaagt ttctgtattt 240 gatgcaattt ccgctgaaaa gatacaagaa attatgctta ttgctgctac tgtggctgct 300 gctgatgttg gctctgttga catgaagaat gctgcaaccg attatgccac catttcacct 360 gctcttacaa ggtgtccttc acttcaaagt actgcaactg cctcggcttc gccacaggca 420 cagttgtatc ctctccccag gacatctttt agcaaactgc aagctgaact accaattgca 480 aggagacact cacttcagcg ttttcttgag aagcggcggg acaggctggt gaacaagaac 540 ccatatcctg gtccatcaac accaaaaatg gctgatgacg tcaaagctga cgtgagtgcc 600 acaacttcac cagagtctgg ctacttcaag gcatcaccca ttcgtcaaga agatatccaa 660 ccaaaagctc cagctcatgt tgcctaa 687 <210> 46 <211> 894 <212> DNA <213> Artificial sequence <400> 46 atgtcaacgg gagaaatggt ttcccggtca cctctataca agcctctcaa ccagctcacc 60 gaggatgaca tttctcaggt cactcgcgaa gattgccgcc tttacctcaa agaaaaaggg 120 atgcggagac cttcgtggaa caaatcgcag gcgattcagc aggtcatctc actgaaaact 180 cttctagaaa cgacatcgga ttctgaagct gtcgaagctt ccaagaaact tcacgttccc 240 ttcccgcaaa atccgcctcg tttcgtttct gattcaaccg ttcaaccgaa tgaaacgaca 300 cggcataaag ggatctcggt cccggtaaac gaatccgttc ctcgcatccg ttcagatccc 360 tcggaattca aattttctcg cgaaaattcc gttcaaaccg ccgtctctgc taatgattct 420 gtttctccaa gatctgcgag tgtagccaaa gagccatcag gacagatgac aattttttat 480 tgtgggaaag tgaatgtcta tgataatata cctggttgta aggcggaagc aatcttgcag 540 tttgctgcaa gcccagtctc atttccacag gaaactctag ttgatcaaag gacctcgcca 600 ttgtccattc catgccatgt acaggctgca ggtgataaag taagccaacg ttcaccaggg 660 gttatattgt catcaatgca agcagtgaag gttgcagaaa actgtcgatt tcctcgagat 720 gactgcaatg tatcttatga agacagcctt gaaggtccca ctagcagaaa cgcattgttg 780 caaagatatc ttgagaaaaa gaaagacaga tatattcgaa gtatgatgtt ggaccagcat 840 tcagcgatta tagcccatgt tgacatggat ggcgctggag tgccgacaga gtag 894 <210> 47 <211> 759 <212> DNA <213> Artificial sequence <400> 47 atgtttggtt caccggaata tacatgtctg aaacctgcaa gtttgccgga gaaaccactg 60 tttaagcgaa cttgtagctt gttaagtcag tacttgaagg aaaaatgtag ctttggtgat 120 cttactttgg gaatcacatg caataacaac gttgaaaaag ggatgccgga gatagtccgt 180 ccggcgacgg agacgacgac gaccaccatg gatttatttc cgagggatca tgtttctggt 240 gttatgagga actcaagatc catggatttg ttccctcaag gagccggttt ttctgccgat 300 aacggttcga gacgggtcgg ggtcgacttg gagactgaaa acgccgcagc gccgatgacg 360 atcttttact gcggacaagt gattgtgttc aatgatttcc cagctgataa agctaaggaa 420 atcatggctt tagcaagcaa gtgcagctcc gaaaacccga aaacgaacac gtttgtaccc 480 ggtagtccga acgaatccgg cctaggagtt cattcgaatt ccaacgatca ggttcctaat 540 atccgaacca atgtatcaac aagtcgtgaa tgcgttcgtt ccattcccgg cgatctaccc 600 attgctcgaa gagcttcgct tcatcgattc ctcgagaaga gaaaggatag gatcacttca 660 aaagcaccat acccaataaa cggctccgcc ggggcatcac ctccgaaacc cggaaatagc 720 aagccgtggc tcggtttagc tgttgaatca ctacagtag 759 <210> 48 <211> 378 <212> DNA <213> Artificial sequence <400> 48 atgagacgaa actgcaactt ggaactggag ctttttcctt cacgtatgtt tccaggccat 60 cgtcaaaaca tggtggaaga gagcaaaaga agcccacaaa atcaacagca gcagctaaca 120 attttctata atggaagagt ttgcgtttgt gatattacag agcttcaggc aagagctatt 180 ttaatgaggg caaatcaaga aacggatgaa agaataaaaa ctccaacagg atcggaaccg 240 gattcaccga catcgtcatc atcgacatcc ctgtctcgat tatgtagccc taatgctggt 300 ctttccatga agaaatcact tcaaaggttc cttcaaaaaa gaaagaacag aatccaagct 360 acgtcccctt accattaa 378 <210> 49 <211> 363 <212> DNA <213> Artificial sequence <400> 49 atgagacgaa actgcaactt ggagcttcgt ctccatcctt ctagttattc cggcggcgac 60 cgggtggaag agagcagtga aaacccagaa aatcaacagc aacagttaac gattttctac 120 aatggaagag tttgtgtttg tgatgttaca gagatccagg caagagccat tcttatgatt 180 gcaaaccgag aaacggatga acgactaagg actccgaggc agggatcgga accggcttca 240 ccgatggtac actctcagat aaatagtccg aataatggac tttccacaag catgaagaga 300 tcgcttcaac ggttcctcca aaaacgaaag aatcgaatcc aagctacctc tccttaccat 360 taa 363 <210> 50 <211> 813 <212> DNA <213> Artificial sequence <400> 50 atgtctaatt tagggcaaaa atctcctgac agagcaagtt ttgtcaatct tctaagccag 60 tacttgaaag aaaaaaggaa tcttggggat ttcagccttg gaatgacttc aaaaccagat 120 gctaaagggc ttgaaacatc taggcaacaa gcaaaaaaca tgaatttttt gtccaatatg 180 ccgaattgtt ctgagagttc aagaccaaat cttgtggcgt ctacctcaaa tgtgaaatct 240 tcagatttct ttccagagat tggcagtttt tgtgcttcca gttccaagga agataccatt 300 aacaagacag atttcatgaa atcagcagca gtggaaccaa agaatgctca gctgaccata 360 ttttttggtg gccaagtgct tgtatataat gattttcccg cggacaaggt gaaggagatc 420 atggccgtag ctaaccgtgg atggtcaact gcatgtagtg gtgttgttgc cgactcttcc 480 atggaaaaac tgaatgctaa ccttgacaaa attgactaca gcagtcctca tatccctgac 540 ttgaacatca cctctgcaac cgtgaatagt tcagctcaag atccttcggt tgagcgacac 600 caatatgtcg gttcagattt gagaattgca agaagaaact cacttcacaa gttctttgaa 660 aagagaaaag acagggcggc catgagagct ccataccaac tgatcaacca ccaaggttcc 720 ccgcctccac ctaaacccga cgaaaacaag ccatctcacg aggaaggtca atcatcaaaa 780 gaaacaccaa gagacattga tctcaactta tag 813 <210> 51 <211> 1014 <212> DNA <213> Artificial sequence <400> 51 atgtctccgg gagaaacggt ctcccggtca cctctagaca agcctcttaa ccagcttact 60 gaggatgaca tttctcaggt cactcgcgaa gattgccgcc gttacctcaa agaaaaaggg 120 atgcggagac cgtcttggaa caaatcgcag gcgattcagc aggtgatctc gttgaaaact 180 cttctagaaa cgacgtctga ttccgacgcc gttgaagctc ggaagaaact ttaccctccc 240 tgcccggaat atccgcctcg cgtcgtttct gattcaaacg ttctaccgaa ggaaattaca 300 ccgaataacg ggatcttggt tccagttccc gaatccgttc cttgtcccca ttcaaatccc 360 tcgaaatccg atttttccgg cgacaattct ggccgaacgg tcatctctgg aaatgattct 420 gtttctccaa gaattgcagg cgcaccaaaa gagtcagcag gacagatgac aatcttttac 480 tgtgggaaag tgaatgtcta tgatgatatg cctggttgta aggctgaagc aatcttgcag 540 cttgctgcaa gcccagtctc atttcctcac gaaattctag ctgatcaaag gaccacacca 600 tggtccattc catgccattc acaggctgca agtgtcaaaa caaccccatg ctcacaaatg 660 gttatattgc cacctcagca aacagaaaac tgtcaatttc ctcgagaaga gagcaatgca 720 tctcttgaag acagccttga aggacctact cgcagaaaag catcggtgca aagatatctt 780 gagaagaaga aagacaggtt taagaacaag agaaagttgg caatgtcttc atctccgacc 840 ttagacatct acttaaatca agtgggagat cagttttcaa atgagcagtt gaaacaaagt 900 gaaccatatt attctcccca agcagaagtg caacgcatgc ctcttgagtg cagctccatt 960 gaaaatgttg caaagattcc ccgtcttact actgatggaa aaggtgacca gtaa 1014 <210> 52 <211> 1113 <212> DNA <213> Artificial sequence <400> 52 atggagagag attttatcgg tttgaggtcc aaaaatgctg caattacaat caaagtagag 60 ccttccaatg ctcagcctga tgattcagtg ttgttgaggg gttcagggat gcagttgtct 120 ttctcaaaca aggtatctac agttcctcag ttcttgtcgt ttgatggtgc tcgagacgat 180 aaaccgagaa aagccacgca tgacactctg ttgtcatccg gttttatgac tattccagct 240 gctgattcta acaagaaacc gcatcctggc ttgactcaga aacaaggggg aaaccactat 300 gcggctacca cctatggtct ccaacaactc gatggacacc aatcccgcct ttcccacgaa 360 gcaaggatat ttccaagttc aagtcagccg aatcacacga ttacggtttc catgaacact 420 cctcttctcc aaccccatct tgcttccccc ggacagaaca taatcggtca tactataaac 480 ccgcaacctt ttaccggagt tccaatcatg gctgcacctg tatctgttgt tcctccatca 540 agtcctatca tcggtactac cgatcttagg aatgctgcca aatcttctag ggcaccagct 600 caattgacta tcttttatgc tggatctgta tgtgtgtacg atgatgtatc tcccgacaag 660 gcccaagcta ttatgttact tgctggaaac gtttcttcta cgactcaaag taaaacggca 720 cctgtgactc aaccgcagac acatattcca cgaccttgta ccttgtcccc tttctcgggt 780 cttccaaatc acctttctgt aacctcccat gtcagtttac agcctgttgc aggatctagt 840 ggcaccaatg aactaacagc tgcaacaagg ataggagctt tagcatctac taataaccaa 900 ccagatcctc caaaattggt taatcctgca gttgctgtgc ctcaggctcg taaagcatcc 960 ttggctcgat ttttagagaa gcgtaaagaa agggtatcaa acacctcacc gtacaacatc 1020 tgcaaaagat ctgaatctgg ccctcttgca tctgatggca taagtttttc cgtgacttct 1080 gctggatcta gtcctttaca agccatcaat taa 1113 <210> 53 <211> 621 <212> DNA <213> Artificial sequence <400> 53 atgtccagag ctaccgtcga gcttgatttc ttcggcatgg agaaagccaa ttcctgcaaa 60 tctcagttcc agaaattcct cgatcgccgc cggagcttcc gaggtattca aggtgccatt 120 tcgaagatga atccggaact tatcaaatct gtgattgctt ccggttcgac gaaccggaat 180 ccggttgatt ggagaaaatc cttttcggtt ccgtcgagtc ctaaagaaga tcggagtact 240 tcgcttcctt ctttacctct actcaatcct gctttgaggt ctactccttc tgaggatagt 300 cctgaaacag caccgttgac gattttttac aacggaacgg tttccgtaat caatgtacct 360 cgagacaagg cggaaagcat cttcaaactt gccgttgaag gaagctcaaa aaacattgaa 420 tctgtagact catcaaaagc tgcaaatcct tcaagtgatc aacaaaacct cctcgaagct 480 cgtaatggag atttgccaat tgctagaaga aagtcgctgc aaagatttct ggaaaagcgc 540 aaagagaggt tagatatatt actgtgttgg agacgaattt acagaatttt tactaatttt 600 ctaaaaaaaa acaacaactg a 621 <210> 54 <211> 1089 <212> DNA <213> Artificial sequence <400> 54 atggagagag attttctggg tttgaattca aatgaaccat tggctgtggt taaggatgat 60 gttaacactg ataagtacaa agaaataggt tttactaaaa gctcagggat acaatggccc 120 ttttcaaaca aagttttcgc tgttccacag ctgatgaact ttaattttgc ccaaggggat 180 aaaaccaaaa agactggata tgattcaaaa gtatcccctc tcttcatgcc tatatcaaca 240 atggatgctg ctgaacttca gaaatcgttc aatcacaaat ggaatggtgg gaatcatttt 300 tcattgactg attctcatgt tcaacacaac acgaacatgt ttccggcatc aaaccagaca 360 atttctgttt ctgggagcga cccattcgtg aagaatcatt acactaccac tggtcaaaaa 420 tttcctgcta atactataaa accacaattt tttggagggg ttccagtgac aactccacat 480 tcagttcttc ccacccttgg ctctgttggt ggaagtgttg aaccatgcac caaggcctct 540 ggatcacctg ctcaattgac tatcttttat gcgggtgaag tgaatgtctt tgatgatatt 600 acccctgaga aggctcaagc tatcatgttt ttggctggga acgggtcttc catggcttct 660 aattcagctt atccaaaacc cccggtccag actcctattt tgaaaccagt tcaagttgat 720 agtgtgcctg cgaaccagct cataaataca caactgagct tcggtaagcc aagcccttta 780 cctgtttcat ctcatgctgg tacgcagtct tggagtgggt ccactagtac cgaagaacag 840 ataatatgca aggcctcagt accttcaact ccatctaccc ctattagcaa attggagagt 900 ccaaatttgg tgaataccat gggatctgat gctgccaccg gcatgatgcc ttctgttcca 960 caggctcgca aagcttcctt ggctcggttt ttggagaagc gcaaggggag gataatgagt 1020 acagcatcac catacaatct tagcaagaag tctctagatt atgcaactac catggaatcg 1080 aatgcataa 1089 <210> 55 <211> 423 <212> DNA <213> Artificial sequence <400> 55 atgtctcaca acattgattt gtttagaaaa tatcttctca gcaaatctga aaaccatacc 60 gttggtaaaa ccaacaagga agaatcagct gtgatgaagc acaggtcctc accacctctt 120 actcctttaa taatgccacc taagtttgct tccagcattt ctcggcccct ttctcttctc 180 gaacgacggc tccttcaacc aaccaggtat ttacaaaaat cttcaacaac acaactcaca 240 attttctacg ctggggttgt taacgtgtat gacaatgttc ccactgataa ggcacaagcc 300 atcatgcttc tggcgggaga gagttgtttg acaaagccta ccgcaaaaga aaagacttta 360 gtagaggcaa aagcatcacc caaccaacaa caagccgtgg gtaataactt tgatagacat 420 taa 423 <210> 56 <211> 585 <212> DNA <213> Artificial sequence <400> 56 atgtcgagag ctagcgtcga gcttgatttc tttggaatgg aggaacagat ctgctgcaaa 60 tctcggtttc agaaatccct cgatcgccgt ctaagctttc gaggtcttca aggcgccctt 120 tcgaaggtga atcctgagct tatcaaatcc gtgattgcct ctggtttgaa gaacccgcag 180 ggtcaggata atgtgtatca gatggactcc aacaagtctt tttcggtccc ttccagtcct 240 aaagaaactc agagtctgtt tccagcttta cctcttctta cccctgctgc cagggctaca 300 tccgagaatg gtcctgaaac agctccctta accattttct acaacggaac ggtttccgtt 360 tttaatgtac ctcgagataa ggcggagagc atcttaaaac tcgcggttga agttgaagga 420 aactcaaaaa atgttgaacc aatagactca aaagtggcaa gtcctcctag cgatcgacaa 480 cagcttcttg aaactctcaa tggagatttg ccaatcgccc ggaggaagtc tttgcagaga 540 tttctggaaa agcgcaaaga gaggtatttt ttgttttgtt tttaa 585 <210> 57 <211> 723 <212> DNA <213> Artificial sequence <400> 57 atgtcgtctt gctcggaatc tacggctatg aaaccggcta ggtcactgga gaagccaagt 60 tttgcccaaa cttgtaattt gttgagtcag tacttgaagg agaaaggtag ctttgggggt 120 ttatctctgg gaatgacatg caacgttgaa gctaatggga caccagtggt gccgcctccg 180 accatgaatc tgttccctgt caatgacaag tccgataatg tttgtggccg aaacggtggg 240 aatcctaaga acttggcatc catggatttg ttcccccaac aagctggttt ggcttctaaa 300 gatgatagtc ccaacaagtt ggagcctcag actgcaccaa tgaccatctt ttatggtgga 360 cgagtgattg tgttcaatga ttttccagca aacaaagcta aggaaatcat gcttttagct 420 agcaacagca gctcacaaag caacgacagc ttcaacccca tcccttttac ttctagcata 480 gccagaagtc caatcaaatc aagcatcgga gttcctccta cctcaaaacc ggttcaccct 540 gctcagcgag ccgtccctgg cgatctgcca attgcaagga gagcttcact gcatcggttc 600 ctagagaagc gaaaggatag gatcaccgca aaagcgccat accagataag tcactccgct 660 gcagctcctt ccctgtccgg cgacaacaag tcatggctcg gtttggctgc tcaatcacca 720 tag 723 <210> 58 <211> 681 <212> DNA <213> Artificial sequence <400> 58 atggaagggg aagctggttc atacgaggac gttaagccaa acgtcgtcgc taaacaaagt 60 aatggcgatg ttgtcgggga taatggtgtt ggaaatctgg gatccgttga agcccctgat 120 tttctttcca agaaaaattt ccagaactgt tccgtggcaa ctcctgcatc tggactgaat 180 tccactggcc ctgctccatc tcaactaacc atcttctatg atggacacgt ttgtgtgttt 240 gatgcaattc ctgtggaaaa ggtgcgggag attatgctta ttgctgcaac tggtgctgct 300 aactctgttg acatgaagaa agttgcaact gattgtgcca ccacctcacc tgttcttacg 360 aggtctcctt cgcttcaaag tactgctact gcaactgctc tggcttcacc acaggcacag 420 gtgtatccta ctaacaggac ccctttctgc aaactgaaag aactaccaat tgcaaggaga 480 cactcccttc agcgattttt tgagaagcgg cgggacaggc tggtgaacag gaatccatat 540 cctaatccat caacaccaaa atcattcgat gacaccaaag ctaacctcag tgctgcaact 600 tcatcagaat cgggttgctt tggtaaatca cctgttgctc aagaagaatt ccatccaaaa 660 gctccagctc atgttgcata a 681 <210> 59 <211> 681 <212> DNA <213> Artificial sequence <400> 59 atgacgaaga actttttgcc caacatggat aattcttatg agagtttaaa accgaatctc 60 ggggcatcta catcgaaatc aaatgcgaaa ccttgtggtt ttttcccaga gattaggagt 120 tttggtgcct ccagttccaa ggaagatacc aacatgatga cagatttcag aaaaccagca 180 aaagtggaac caaagaattc ccagatgacc atattttttg gtggtcaagt ggcagtattt 240 aatgacttcc cagctgacaa gttcaaggaa atcatggact tattagctag ccatggatgc 300 tcaaccgcaa gcggtgttgt tgttgatact gtcatggaaa aagttaagtc taaaacagtc 360 caaatcgagc ctagcaatca tgaaattcca gacctgaatg tttctaccgc gactgggaat 420 agtcctcctc cccctcatga ttcttccgtt gaatggcatc aatatggcgg ttcaggacct 480 tcagatcttc gaattgcaag aagaaattca cttcacaagt ttttcgaaaa gagaaaagaa 540 agggcaacag ctagagcgcc ataccaagtt aacaacgcaa gaggatcgac tctgccacct 600 aaacccgatg aaaacaaatc atctcacgag gaaggtcaat catcgaaaga agcctcaaga 660 gatcttgatc tcaagttata g 681 <210> 60 <211> 1098 <212> DNA <213> Artificial sequence <400> 60 atggagagag attttctggg tttgaactca aaacaatcct tccctttggt taaagaagaa 60 gttgaagaaa ttggttttac caaaagcttg gggattcaat ggccaatttc aaacaaggtt 120 tcttcagctg ttccaccaca gcagatgagt tttgattttg ctcaaggaga taacgctaaa 180 agaatgggat atgattcaat agtatcctct actttcatgc atatctcacc accagatgct 240 gctgaacttc agaaatcctt caatcacaac aggcaacgag taggcaatca tttcccattc 300 acagctgctt cttctgttca gcatgatgta catcatgttc aacggcctta tgacatgaaa 360 atgtttccgg tctcgaacca atcagttccc gtttcaacta ccaacccatt tatgaatagt 420 catttcacta ctactgctat gaaatcacaa ttgcttggag gaattccagt tacgactccg 480 ccgcattcgg ttcttcccac cctcggttcc tttggtggaa gcatcgaacc acggaaaagt 540 gtcaggggcc ttggggactc cagatcacct gttcaattga ctatctttta tgctggtacc 600 gtgaatgttt atgatgatat cacccccgag aaggctcagg ctatcatgtt acttgctggg 660 aatggatctt ccttgacttc taatgtggca catccaaatg tccaagtcca ggcaccaatt 720 tcaaaaccac ttcaattcga aactcttccc acgaatcatt tcacgaatgc agaactttgt 780 tctggtatcc caagcccttt atctgtttca tctcacactg gtgttcaatc caggagtgga 840 tccactagta ctgatgaaaa gaccgtatgc aagaccacag gaagtctgac aacccctatt 900 agcctagtgg agtctcctaa attggcaaat accatgggac ctgttacccc aaccagcatg 960 atgccttctg tgccccaggc tcgcaaagca tccttggctt ggtttttgga gaagcgcaag 1020 gaaaggatta tgactgcatc accatatgat ctaagcaaga aaccaccaca ttgtacaacc 1080 cagggatcaa atgcatga 1098 <210> 61 <211> 360 <212> DNA <213> Artificial sequence <400> 61 atgagacgaa actgcaactt ggaacttcgt cttcttcctt ctagttatcc cagcgacagt 60 cacgacatga tggaagcaag aattgaaagc ccagaaaccc agcagcaaca gctaacaatt 120 ttctacaatg gaagagtttg cgtatctgat gttacagagc ttcaggcaaa agccattcta 180 atattagcaa accgggaaag agatgaaaga atgaaaagtc cgacgggatg tgaaccggtg 240 tcgccgacgt taaaatccca ggtgaatagc ccaaacacat cactttccat gaagagatcg 300 cttcaacggt tcctccagaa acgaaagacg cgaatccaag ctacctctcc ttaccattaa 360 <210> 62 <211> 792 <212> DNA <213> Artificial sequence <400> 62 atgaatatgt cgtgttcacc ggaatttatg gttcagaaac cggcgaggtc accggagaag 60 acaagtttta cccaaacttg taacttattg agtcagtact tgaaggagaa aggtagcttt 120 ggagatctaa gtctgggtat gacatgcaac gttgaagcta atgagactcc cgagatgctg 180 cgtcccacca tgaatttgtt tcctgtcaat gggaaatccg gtgatgattg ccatgcggcg 240 cctcctcctc ggaaactgag atccatggat ttgttcccta atcaagctgc tttttcatca 300 cctaaagatg atgctctgaa aagcaccatg aacaaattgg gttcttcagt ggagcctcaa 360 actgcacaga tgaccatctt ttatggtgga caagtgattg tgttcaacga tttcccggcc 420 gacaaagcta aggagatcat gcttttagct ggcaaaggca gctctcaaaa caatagcttc 480 aaccccaatc ctccccacat caatgccccc tttacttcta ccatagcaac aagtccgatt 540 gagtccggta tcggggttcc tcctacccct aatttcagca ccacggtaac tcaagaatgc 600 atacgatctg ctcagcgacc cattcccggc gatctaccaa ttgcaaggag agcttcactc 660 caccggttcc ttgagaagag aaaggatagg atgaccacaa gtgcaccata ccagataagt 720 aactccacag catcttcatc caagccaggc aacgacaagt catggctcgg tttggctgct 780 caatctccgt ag 792 <210> 63 <211> 687 <212> DNA <213> Artificial sequence <400> 63 atggaagaag aagctgaatc acgcgaggaa gtcaagccta acgtcgtcgt taaagagact 60 aatggagata ttgtcggcga taacgatgca ggaaaattgg gtaccattga aacccctgat 120 tttctttccc agaaaatatc ccataactgt tccctaccga ttctggcatc tggagtaaat 180 accacaagcc ttgctcaatc ccaactcacc atcttctatg gcggaaaagt ttctgtattt 240 gatgcaattt ccgctgaaaa gattcaagaa attatgctta ttgctgctgc tgtggctgct 300 gctgatgttg gctctgttga catgaagaat gctgcaaccg attatgccac catttcacct 360 gctcttacaa ggtgtccttc acttcaaagt actgcaactg ctttggcttc gccacaggca 420 cagttgtatc ctttccccag gacatctttt agcaaactgc aagctgaact accaattgca 480 aggagacact cacttcagcg ttttcttgag aagcggcggg acaggctggt gaacaagaac 540 ccatatcctg gtccatcaac accaaaaatg gttgatggcg ccaaagctga cgtgagtgcc 600 acaacttcac cagagtctgg ctgcttcaag gcatcaccca ttcgtcaaga agatatccaa 660 ccaaaagctc cagctcatgt tgcgtaa 687 <210> 64 <211> 672 <212> DNA <213> Artificial sequence <400> 64 atgtttggtt caccggagaa accaatgttt aagcgaactt gtagcttgtt aagtcagtac 60 ttgaaggaaa aaggtagctt tggtgatctt actttgggaa tcacatgcag tcacaacgtt 120 gaaaaaggga tgccggagat agtccgtccg gcgacggaga cgaggacgac caccatggat 180 ttgttccctc aaggagccgg tttttctgcc gataacggtt cgagacgggt cggggtcgat 240 gtggaccctc aaaacgcggc agcgccgatg acgatctttt actgcggaca agtgattgtg 300 ttcaatgatt tcccagctga taaagctaag gaaatcatgg ctttagcaag caagtgcagc 360 tccgaaaacc cgaaaacgaa cacgtttgta cccggtagtc cgaacgaatc cggcctagga 420 gttcattcga attccaacga tcaggttcct aatatccgaa ccaatttacc aacaagtcgc 480 gaatgcgttc gtcccattcc cggcgatcta cccattgctc gaagagcttc gcttcaccga 540 ttcctcgaga agagaaagga taggatcact tcaaaagcac catacccaat aaacggctcc 600 gccggggcat cgccgccgaa acccggaaat agcaagccgt ggctcggttt agctgttgaa 660 tcactgcagt ag 672 <210> 65 <211> 378 <212> DNA <213> Artificial sequence <400> 65 atgagacgaa agtgcaactt ggaactggag ctttttcctt cacgtatttt tccaggccat 60 cgtcaaaaca tggtggaaga gagcaaaaga atcccacaaa atcaacagca gcagctaacc 120 attttctata atggaagagt ttgcgtttgt gatgttacag agcttcaggc aagagctatt 180 ttaatgaggg caaatcgaga agcggatgaa agaataaaaa ctccaacagg atcggaaccg 240 gattcaccga catcgtcatc atcgacatcc ccgtctcgat tatgtagccc taatgctggt 300 ctttccatga agaaatccct tcaaaggttc cttcaaaaaa gaaagaacag aatccaagct 360 acctcccctt accattaa 378 <210> 66 <211> 363 <212> DNA <213> Artificial sequence <400> 66 atgaggcgaa actgcaactt ggagcttcgt ctccatcctt ctagttattc cggcggcgac 60 cgggtggaag agagcagtga aaacccagaa aatcaacagc aacagttaac gattttctac 120 aatggaagag tttgtgtttg tgatgttaca gagatccagg caagagccat tcttatgatt 180 gcaaaccgag aaacggatga acgattaagg actccgaggt cgggatcggg accggcttcg 240 ccgacggtac actctcagat aaatagtccg aataatggac tttccacaag catgaagaga 300 tcgcttcaac ggttcctcca aaaacgaaag aatcgaatcc aagctacctc tccttaccat 360 taa 363 <210> 67 <211> 813 <212> DNA <213> Artificial sequence <400> 67 atgtctaatt tagggcaaaa atctcctgac agagcaagtt ttgtcaatct tctaagccag 60 tacttgaaag aaaaaaggaa tcttggggat ttcagccttg gaatgacttc aaaaccagat 120 gctaaagggc ttgaaacatc taggcaacat gcaaaaaaca tgaatttttt gtccaatatg 180 ccgaattgtt ctgagagttc aagaccaaat cttgtggcat ctacctcaaa tgtgaaatct 240 tcagatttct ttccagagat tggcagtttt ggtgcctcca gttccaagga agataccttt 300 aacaagacag atttcatgaa atcagcagca gtggaaccaa agaatgctca gctgaccata 360 ttttttggtg gccaagtgtt tgtatataat gattttcccg cggacaaggt gaaggagatc 420 atggccgtag ctaaccgtgg atggtcaacc tcatgtagtg gtgttgttgc cgactctgcc 480 atggaaaaac tgaatgctaa ccttgacaaa attgactaca gcagtcctca tatccctgac 540 ttgaacatca cctctgcaac cgctaatagt ccagctcaag atccttcggt cgagcgatgc 600 caatatgtcg gttcagattt gagaattgca agaagaaact cacttcacaa gttctttgaa 660 aagagaaaag acagggcggc catgagagca ccataccaac tgaacaacca ccaaggttcc 720 ccgcctccac ctaagcccga cgaaaacaag ccatctcacg aggaaggtca atcatcaaaa 780 gaaacgccaa gagacattga tctcaactta tag 813 <210> 68 <211> 1098 <212> DNA <213> Artificial sequence <400> 68 atggaggctg gggtaacgac gacggcgact acaacagcgt cgttcagttc gatacttgat 60 aaacccctca gccaactaac cgaagaagac atttctcaac tcactcgcga agactgtcgc 120 aaattcctca aagaaaaagg aatgcgtagg ccgtcatgga acaaatcgca ggcgatccag 180 caagtgattt cgttcaaggc gttgttggaa agcaacgaag attccggcgc cggagctcgc 240 cggaaaatcc ttgtttgtcc accaccgtca cattttcctc cgcaaaatgc ggtagcttcc 300 aattctggtg agtcagtaaa agaagcagtc tttggagaag aagaaagcct gtacggccaa 360 aaagatcttt ctttgaaagc tgctccggtg gtgcagatga attgtcaggg cggtgacacg 420 gatgacaaga ctctttcgcc tagtttaggc tctccacggg agtattcaaa attgcctggc 480 agaagtcaat gtgaaacaaa tgagttgggt gggcaaatga caatttttta ctgtggaaag 540 atcaatgtgt acgatggtgt accacttgct aaggcacgag caatcatgca cctggcagct 600 tctcctattg attttcctca gggcaatcta tgtaatcaaa atggcgcctt taggtccttt 660 ctgggtcatg tacaagaagc cgaagacaaa aacgacctta cttcatctat tgctttgaac 720 ttgaattctc atatcatgca cactgagaag atgacagaat atcagcagca gtttagggga 780 aaagcaaaca tcagtcgtga ttctgatgta gatggacagg tgagcagaaa agtatcattg 840 cagcgatatc ttgaaaagcg aaaagacagg ggaagattct ttaaaggcag gaaaaatgca 900 ggacaaactt tgtctagctc ggagatgtac ctgaaccatc agataagagc tcactactta 960 aatggacaaa caaaccagag cagaacaagt tctccaccac agtctggagt gccacatgca 1020 ttttatagct cagctgacaa ccaagagctt gtgaattttt ctgtagatct caatgatgaa 1080 ggtggtcaag aacactga 1098 <210> 69 <211> 1014 <212> DNA <213> Artificial sequence <400> 69 atgtctccgg gagaaacggt ctcccggtca cctctagaca agcctctgaa ccagcttact 60 gaggatgaca tttctcaggt cactcgtgaa gattgccgcc gttacctcaa agaaaaaggg 120 atgcggagac cgtcttggaa caaatcgcag gcgattcagc aggtgatctc gttgaaaact 180 cttctagaaa cgacgtctga ttccgacgcc gttgaagctc ggaagaaact ttaccctccc 240 tgcccggaat atccgcctcg cgtcgtttct gattcaaacg ttctaccgag ggaaatgaca 300 ccgaataacg ggatattggt tccagtttcc gaatccgttc cttgtcccct ttcaaatccc 360 tcgaaatccg atttttccgg cgacaattct ggccgaacgg tcatctccgg aaatgattct 420 gtttctccaa gaattgcagg tgcagcaaaa gagccagcag gacagatgac aatcttttac 480 tgtgggaaag tgaatgtcta tgatgatatg cctggttgta aggctgaagc aatcatgcag 540 cttgctgcaa gcccagtctc atttcctcac gaaattctag ctgatcaaag gtccacacca 600 tggtccattc catgccattc acaggctgca agtgtcaaaa caaccccatg ttcacaaatg 660 gttatattgc cacctcagca aacagaaaac tgtcaatttc ctcgagaaga gagcaatgca 720 tctcttgaag acagccttga aggacctact agcagaaaag cattggtgca tagatatctt 780 gagaagaaga aagacaggtt taagaacaag agaaagttgg caatgtcttc atctccgacc 840 ttagacatct acttaaatca agtgggagat cagttttcaa atgagcagtt gaaacaaagt 900 gaaccatatt attctcccca agcagaagca caccgcatgc ctcttgagtg cagctccatt 960 gaaaatgttg caaagattcc ccgccttact actgatggaa aaggtaaccg gtaa 1014 <210> 70 <211> 1116 <212> DNA <213> Artificial sequence <400> 70 atggagagag attttctcgg tttgaggtcc aaaaatgctg caattacaat caaagtagag 60 ccctccgatg ctcagcctaa tgattcagtt tggttgaggg gttcagggat gcagttgtct 120 ttcacaaaca aggtatctac agttcctcag ttcttgtcgt tcgatggtgc tcgagacgat 180 aaaccaagaa aagccacgca tgacacgctg ttgtcatccg gttttatgac tattccaact 240 actgattcta acaagaaacc gcatcctggc ttgactcaga aacaaggggg aaaccactat 300 gcggctacca cctatggtct ccaacaactc gatgtacacc aatcccgcct ttcccacgaa 360 gcaaggatat ttccaagttc aagtcagctg aatcacacga ttacggtttc tatgaacact 420 cctcttctcc aaccccatct tgcttccccc ggacagaaca taatcggtca tactataaac 480 ccgcaacctt ttaccggagt tccaatcatg gctgcacctg tatctgttgt tcctccatca 540 agtcctatca tcggtaccac cgatcttagg aatgctgcca aatcttctaa ggcaccagct 600 cagttgacta tcttctatgc tggatctgta tgtgtgtacg atgatgtatc tcccgacaag 660 gcccaagcta ttatgttact tgctggaaac ggttcttctg caactcaaag taaaacggca 720 cctgtgactc aaaggcagac acatattcca cgaccttgta ccttgtcccc tttctcgggt 780 cttccaaatc acctttctgc aacctcccat gtcagtttac agcctgttgc aggatctagt 840 ggcaccaatg aactaacagc tgcaacaagg ataggagctt tagcatccgc taataaccaa 900 ccagatcctc ccaaattggt taatcctgca gttgctgtgc ctcaggctcg taaagcatcc 960 ttggctcggt ttttagagaa gcgtaaagaa agggtatcaa acacctcacc gtacaacatc 1020 tgcaaaagat ctcctgaatc tggccctctt gcatctgatg gcataagttt tcctgtgact 1080 tctgctggat ctagtccttt acaagccatc aattaa 1116 <210> 71 <211> 490 <212> PRT <213> Artificial sequence <400> 71 Met His His His His His His Glu Asn Leu Tyr Phe Gln Gly Met Ser 1 5 10 15 Ser Cys Ser Glu Ser Thr Ala Met Lys Pro Ala Arg Ser Pro Glu Lys 20 25 30 Pro Ser Phe Ala Gln Thr Cys Asn Leu Leu Ser Gln Tyr Leu Lys Glu 35 40 45 Lys Gly Ser Phe Gly Asp Leu Ser Leu Gly Met Thr Cys Asn Val Glu 50 55 60 Ala Asn Gly Thr Pro Val Val Pro Pro Pro Thr Met Asn Leu Phe Pro 65 70 75 80 Leu Asn Asp Lys Ser Asp Asp Val Cys Gly Arg Asn Gly Gly Asn Pro 85 90 95 Lys Asn Leu Thr Ser Met Asp Leu Phe Pro Gln Gln Ala Gly Leu Ala 100 105 110 Ser Lys Asp Asp Ser Pro Asn Lys Leu Glu Pro Gln Thr Ala Pro Met 115 120 125 Thr Ile Phe Tyr Gly Gly Gln Val Ile Val Phe Asn Asp Phe Pro Ala 130 135 140 Asn Lys Ala Lys Glu Ile Met Leu Leu Ala Ser Asn Ser Ser Ser Gln 145 150 155 160 Ser Asn Asn Ser Phe Asn Pro Ile Pro Phe Thr Ser Ser Ile Ala Arg 165 170 175 Ser Pro Ile Glu Ser Ser Ile Gly Val Pro Pro Thr Ser Lys Pro Val 180 185 190 His Pro Ala Gln Arg Ala Val Pro Gly Asp Leu Pro Ile Ala Arg Arg 195 200 205 Ala Ser Leu His Arg Phe Leu Glu Lys Arg Lys Asp Arg Ile Thr Ala 210 215 220 Lys Ala Pro Tyr Gln Ile Asn Asn Ser Ala Ala Ala Pro Ser Leu Ser 225 230 235 240 Gly Asp Asn Lys Ser Trp Leu Gly Leu Ala Ala Gln Ser Pro Met Val 245 250 255 Ser Lys Gly Glu Glu Asp Asn Met Ala Ile Ile Lys Glu Phe Met Arg 260 265 270 Phe Lys Val His Met Glu Gly Ser Val Asn Gly His Glu Phe Glu Ile 275 280 285 Glu Gly Glu Gly Glu Gly Arg Pro Tyr Glu Gly Thr Gln Thr Ala Lys 290 295 300 Leu Lys Val Thr Lys Gly Gly Pro Leu Pro Phe Ala Trp Asp Ile Leu 305 310 315 320 Ser Pro Gln Phe Met Tyr Gly Ser Lys Ala Tyr Val Lys His Pro Ala 325 330 335 Asp Ile Pro Asp Tyr Leu Lys Leu Ser Phe Pro Glu Gly Phe Lys Trp 340 345 350 Glu Arg Val Met Asn Phe Glu Asp Gly Gly Val Val Thr Val Thr Gln 355 360 365 Asp Ser Ser Leu Gln Asp Gly Glu Phe Ile Tyr Lys Val Lys Leu Arg 370 375 380 Gly Thr Asn Phe Pro Ser Asp Gly Pro Val Met Gln Lys Lys Thr Met 385 390 395 400 Gly Trp Glu Ala Ser Ser Glu Arg Met Tyr Pro Glu Asp Gly Ala Leu 405 410 415 Lys Gly Glu Ile Lys Gln Arg Leu Lys Leu Lys Asp Gly Gly His Tyr 420 425 430 Asp Ala Glu Val Lys Thr Thr Tyr Lys Ala Lys Lys Pro Val Gln Leu 435 440 445 Pro Gly Ala Tyr Asn Val Asn Ile Lys Leu Asp Ile Thr Ser His Asn 450 455 460 Glu Asp Tyr Thr Ile Val Glu Gln Tyr Glu Arg Ala Glu Gly Arg His 465 470 475 480 Ser Thr Gly Gly Met Asp Glu Leu Tyr Lys 485 490 <210> 72 <211> 1470 <212> DNA <213> Artificial sequence <400> 72 atgcatcacc accaccacca cgaaaaccta tacttccaag gaatgtcaag ctgcagcgag 60 agcaccgcga tgaagccggc gcgtagcccg gaaaaaccga gcttcgcgca gacctgcaac 120 ctgctgagcc aatatctgaa ggagaaaggc agctttggtg acctgagcct gggcatgacc 180 tgcaacgttg aagcgaacgg taccccggtg gtgccgccgc cgaccatgaa cctgttcccg 240 ctgaacgaca agagcgacga tgtgtgcggc cgtaacggtg gcaacccgaa aaacctgacc 300 agcatggacc tgttcccgca gcaagcgggt ctggcgagca aggacgatag cccgaacaaa 360 ctggagccgc agaccgcgcc gatgaccatc ttctacggtg gccaagtgat tgttttcaac 420 gattttccgg cgaacaaggc gaaagaaatc atgctgctgg cgagcaacag cagcagccaa 480 agcaacaaca gcttcaaccc gatcccgttt accagcagca ttgcgcgtag cccgatcgag 540 agcagcatcg gtgtgccgcc gaccagcaaa ccggttcacc cggcgcaacg tgcggttccg 600 ggtgacctgc cgattgcgcg tcgtgcgagc ctgcaccgtt ttctggaaaa gcgtaaagat 660 cgtatcaccg cgaaagcgcc gtaccagatt aacaacagcg cggcggcgcc gagcctgagc 720 ggtgacaaca aaagctggct gggtctggcg gcgcaaagcc cgatggttag caaaggcgag 780 gaagataaca tggcgatcat taaggagttc atgcgtttta aagtgcacat ggaaggcagc 840 gttaacggtc acgagttcga aatcgagggt gaaggcgagg gtcgtccgta cgagggtacc 900 cagaccgcga agctgaaagt taccaagggt ggcccgctgc cgtttgcgtg ggacatcctg 960 agcccgcaat tttgtacgg cagcaaggcg tatgtgaaac acccggcgga cattccggat 1020 tatctgaagc tgagcttccc ggagggtttt aaatgggaac gtgttatgaa ctttgaggat 1080 ggtggcgtgg ttaccgtgac ccaggacagc agcctgcaag atggcgaatt catctacaag 1140 gttaaactgc gtggcaccaa ctttccgagc gacggtccgg tgatgcagaa gaaaaccatg 1200 ggttgggagg cgagcagcga acgtatgtat ccggaggatg gcgcgctgaa gggtgaaatt 1260 aaacagcgtc tgaagctgaa agacggtggc cactacgatg cggaagttaa gaccacctat 1320 aaagcgaaga aaccggttca actgccgggc gcgtacaacg tgaacatcaa gctggacatt 1380 accagccaca acgaggatta caccatcgtg gaacagtatg agcgtgcgga aggtcgtcac 1440 agcaccggtg gcatggacga actgtataaa 1470 <210> 73 <211> 207 <212> PRT <213> Artificial sequence <400> 73 Master Sister Sister Cys Sister Glu Sister Thr Here Master Lys Pro Here Arg Sister Pro 1 5 10 15 Glu Ala Asn Gly Thr Pro Val Val Pro Pro Pro Thr Met Asn Leu Phe 20 25 30 Pro Leu Asn Asp Lys Ser Asp Asp Val Cys Gly Arg Asn Gly Gly Asn 35 40 45 Pro Lys Asn Leu Thr Ser Met Asp Leu Phe Pro Gln Gln Ala Gly Leu 50 55 60 Ala Ser Lys Asp Asp Ser Pro Asn Lys Leu Glu Pro Gln Thr Ala Pro 65 70 75 80 Met Thr Ile Phe Tyr Gly Gly Gln Val Ile Val Phe Asn Asp Phe Pro 85 90 95 Ala Asn Lys Ala Lys Glu Ile Met Leu Leu Ala Ser Asn Ser Ser Ser 100 105 110 Gln Ser Asn Asn Ser Phe Asn Pro Ile Pro Phe Thr Ser Ser Ile Ala 115 120 125 Arg Ser Pro Ile Glu Ser Ser Ile Gly Val Pro Pro Thr Ser Lys Pro 130 135 140 Val His Pro Ala Gln Arg Ala Val Pro Gly Asp Leu Pro Ile Ala Arg 145 150 155 160 Arg Ala Ser Leu His Arg Phe Leu Glu Lys Arg Lys Asp Arg Ile Thr 165 170 175 Ala Lys Ala Pro Tyr Gln Ile Asn Asn Ser Ala Ala Ala Pro Ser Leu 180 185 190 Ser Gly Asp Asn Lys Ser Trp Leu Gly Leu Ala Ala Gln Ser Pro 195 200 205 <210> 74 <211> 624 <212> DNA <213> Artificial sequence <400> 74 atgtcgtctt gctcggaatc tacggctatg aaaccggcta ggtcaccgga ggctaatggg 60 acaccagtgg tgccgcctcc gaccatgaat ctgttccctc tcaatgacaa gtccgatgat 120 gtttgtggcc gaaacggtgg gaatcctaag aacttgacat ccatggattt gttcccccaa 180 caagctggtt tggcttctaa agatgatagt cccaacaagt tggagcctca gactgcacca 240 atgaccatct tttatggtgg acaagtgatt gtgttcaatg attttccagc aaacaaagct 300 aaggaaatca tgcttttagc tagcaacagc agctcacaaa gcaacaacag cttcaacccc 360 atccctttta cttctagcat agccagaagt ccaatcgaat caagcattgg agttcctcct 420 acctcaaaac cggttcaccc tgctcagcga gctgtccctg gcgatctgcc aattgcaagg 480 agagcttcac tgcatcggtt cctagagaag cgaaaggata ggatcactgc aaaagcgcca 540 taccagataa ataactcagc tgcagctcct tccctgtccg gcgataacaa gtcatggctc 600 ggtttggctg ctcaatcacc atag 624 <210> 75 <211> 210 <212> PRT <213> Artificial sequence <400> 75 Met Ser Ser Cys Ser Glu Ser Thr Ala Met Lys Pro Ala Arg Ser Pro 1 5 10 15 Glu Lys Pro Ser Phe Ala Gln Thr Cys Asn Leu Leu Ser Gln Tyr Leu 20 25 30 Lys Glu Lys Gly Ser Phe Gly Asp Leu Ser Leu Gly Met Thr Cys Asn 35 40 45 Val Glu Ala Asn Gly Thr Pro Val Val Pro Pro Pro Thr Met Asn Leu 50 55 60 Phe Pro Leu Asn Asp Lys Ser Asp Asp Val Cys Gly Arg Asn Gly Gly 65 70 75 80 Asn Pro Lys Asn Leu Thr Ser Met Asp Leu Phe Pro Gln Gln Ala Gly 85 90 95 Leu Ala Ser Lys Asp Asp Ser Pro Asn Lys Leu Glu Pro Gln Thr Ala 100 105 110 Ser Ser Ser Gln Ser Asn Asn Ser Phe Asn Pro Ile Pro Phe Thr Ser 115 120 125 Ser Ile Ala Arg Ser Pro Ile Glu Ser Ser Ile Gly Val Pro Pro Thr 130 135 140 Ser Lys Pro Val His Pro Ala Gln Arg Ala Val Pro Gly Asp Leu Pro 145 150 155 160 Ile Ala Arg Arg Ala Ser Leu His Arg Phe Leu Glu Lys Arg Lys Asp 165 170 175 Arg Ile Thr Ala Lys Ala Pro Tyr Gln Ile Asn Asn Ser Ala Ala Ala 180 185 190 Pro Ser Leu Ser Gly Asp Asn Lys Ser Trp Leu Gly Leu Ala Ala Gln 195 200 205 Ser Pro 210 <210> 76 <211> 633 <212> DNA <213> Artificial sequence <400> 76 atgtcgtctt gctcggaatc tacggctatg aaaccggcta ggtcaccgga gaagccaagt 60 tttgcccaaa cttgtaattt gttgagccag tacttgaagg agaaaggtag ctttggggat 120 ctatctctgg gatgacatg caacgttgaa gctaatggga caccagtggt gccgcctccg 180 accatgaatc tgttccctct caatgacaag tccgatg ttgtggccg aaacggtggg 240 aatcctaaga acttgacatc catggattg ttccccac aagctggttt ggctctaaa 300 gatgatagtc ccacagtt ggagcctcag actgcaagca gctcacaag 360 ttcacccca tccttttac tttagcata gccagaagtc caatcgaatc aagcattgga 420 gttcctccta cctcaaacc ggttcaccct gctcagcgag ctgtccctgg cgatctgcca 480 attgcaagga gagcttcact gcatcggttc ctagagaagc gaaggatag gatcactgca 540 aaagcgccat accagataaa taactcagct gcagctcctt ccctgtccgg cgataacaag 600 tcatggctcg gttggctgc tcaatcacca tag 633 <210> 77 <211> 211 <212> PRT <213>人工序列(Artificial sequence) <400> 77 Met Ser Ser Cys Ser Glu Ser Thr Ala Met Lys Pro Ala Arg Ser Pro 1 5 10 15 Glu Lys Pro Ser Phe Ala Gln Thr Cys Asn Leu Server Gln Tyr Leu 20 25 30 Lys Glu Lys Gly Ser Phe Gly Asp Leu Ser Leu Gly Met Thr Cys Asn 35 40 45 Val Glu Ala Asn Gly Thr Pro Val Val Pro Pro Pro Thr Met Asn Leu 50 55 60 Phe Pro Leu Asn Asp Lys Ser Asp Asp Val Cys Gly Arg Asn Gly Gly 65 70 75 80 Asn Pro Lys Asn Leu Thr Ser Met Asp Leu Phe Pro Gln Gln Ala Gly 85 90 95 Leu Ala Ser Lys Asp Asp Ser Pro Asn Lys Leu Glu Pro Gln Thr Ala 100 105 110 Pro Met Thr Ile Phe Tyr Gly Gly Gln Val Ile Val Phe Asn Asp Phe 115 120 125 Pro Ala Asn Lys Ala Lys Glu Ile Met Leu Leu Ala Ser Asn Ser Ser 130 135 140 Ser Gln Ser Asn Asn Ser Phe Asn Pro Ile Pro Phe Thr Ser Ser Ile 145 150 155 160 Ala Arg Ser Pro Ile Glu Ser Ser Ile Gly Val Pro Pro Thr Ser Lys 165 170 175 Pro Val His Pro Ala Gln Arg Ala Val Pro Gly Asn Asn Ser Ala Ala 180 185 190 Ala Pro Ser Leu Ser Gly Asp Asn Lys Ser Trp Leu Gly Leu Ala Ala 195 200 205 Gln Ser Pro 210 <210> 78 <211> 636 <212> DNA <213> Artificial sequence <400> 78 atgtcgtctt gctcggaatc tacggctatg aaaccggcta ggtcaccgga gaagccaagt 60 tttgcccaaa cttgtaattt gttgagccag tacttgaagg agaaaggtag ctttggggat 120 ctatctctgg gaatgacatg caacgttgaa gctaatggga caccagtggt gccgcctccg 180 accatgaatc tgttccctct caatgacaag tccgatgatg tttgtggccg aaacggtggg 240 aatcctaaga acttgacatc catggatttg ttcccccaac aagctggttt ggcttctaaa 300 gatgatagtc ccaacaagtt ggagcctcag actgcaccaa tgaccatctt ttatggtgga 360 caagtgattg tgttcaatga ttttccagca aacaaagcta aggaaatcat gcttttagct 420 agcaacagca gctcacaaag caacaacagc ttcaacccca tcccttttac ttctagcata 480 gccagaagtc caatcgaatc aagcattgga gttcctccta cctcaaaacc ggttcaccct 540 gctcagcgag ctgtccctgg caataactca gctgcagctc cttccctgtc cggcgataac 600 aagtcatggc tcggtttggc tgctcaatca ccatag 636 <210> 79 <211> 252 <212> PRT <213> Artificial sequence <400> 79 Met Gly His Pro Pro Leu Leu Pro Leu Leu Leu Leu Leu His Thr Cys 1 5 10 15 Val Pro Ala Ser Trp Gly Leu Arg Cys Met Gln Cys Lys Thr Asn Gly 20 25 30 Asp Cys Arg Val Glu Glu Cys Ala Leu Gly Gln Asp Leu Cys Arg Thr 35 40 45 Thr Ile Val Arg Leu Trp Glu Glu Gly Glu Glu Leu Glu Leu Val Glu 50 55 60 Lys Ser Cys Thr His Ser Glu Lys Thr Asn Arg Thr Leu Ser Tyr Arg 65 70 75 80 Thr Gly Leu Lys Ile Thr Ser Leu Thr Glu Val Val Cys Gly Leu Asp 85 90 95 Leu Cys Asn Gln Gly Asn Ser Gly Arg Ala Val Thr Tyr Ser Arg Ser 100 105 110 Arg Tyr Leu Glu Cys Ile Ser Cys Gly Ser Ser Asp Met Ser Cys Glu 115 120 125 Arg Gly Arg His Gln Ser Leu Gln Cys Arg Ser Pro Glu Glu Gln Cys 130 135 140 Leu Asp Val Val Thr His Trp Ile Gln Glu Gly Glu Glu Gly Arg Pro 145 150 155 160 Lys Asp Asp Arg His Leu Arg Gly Cys Gly Tyr Leu Pro Gly Cys Pro 165 170 175 Gly Ser Asn Gly Phe His Asn Asn Asp Thr Phe His Phe Leu Lys Gly 180 185 190 Asp Ala Phe Ser Met Asn His Ile Asp Val Ser Cys Cys Thr Lys Ser 195 200 205 Gly Cys Asn His Pro Asp Leu Asp Val Gln Tyr Arg Ser Gly Ala Ala 210 215 220 Pro Gln Pro Gly Pro Ala His Leu Ser Leu Thr Ile Thr Leu Leu Met 225 230 235 240 Thr Ala Arg Leu Trp Gly Gly Thr Leu Leu Trp Thr 245 250 <210> 80 <211> 759 <212> DNA <213> Artificial sequence <400> 80 atgggtcacc cgccgctgct gccgctgctg ctgctgctcc acacctgcgt cccagcctct 60 tggggcctgc ggtgcatgca gtgtaagacc aacggggatt gccgtgtgga agagtgcgcc 120 ctgggacagg acctctgcag gaccacgatc gtgcgcttgt gggaagaagg agaagagctg 180 gagctggtgg agaaaagctg tacccactca gagaagacca acaggaccct gagctatcgg 240 actggcttga agatcaccag ccttaccgag gttgtgtgtg ggttagactt gtgcaaccag 300 ggcaactctg gccgggctgt cacctattcc cgaagccgtt acctcgaatg catttcctgt 360 ggctcatcag acatgagctg tgagaggggc cggcaccaga gcctgcagtg ccgcagccct 420 gaagaacagt gcctggatgt ggtgacccac tggatccagg aaggtgaaga agggcgtcca 480 aaggatgacc gccacctccg tggctgtggc taccttcccg gctgcccggg ctccaatggt 540 ttccacaaca acgacacctt ccacttcctg aaaggtgacg ccttcagcat gaaccacatt 600 gatgtctcct gctgtactaa aagtggctgt aaccacccag acctggatgt ccagtaccgc 660 agtggggctg ctcctcagcc tggccctgcc catctcagcc tcaccatcac cctgctaatg 720 actgccagac tgtggggagg cactctcctc tggacctaa 759 <210> 81 <211> 251 <212> PRT <213> Artificial sequence <400> 81 Met Gly His Pro Pro Leu Leu Pro Leu Leu Leu Leu Leu His Thr Cys 1 5 10 15 Val Pro Ala Ser Trp Gly Leu Arg Cys Met Gln Cys Lys Thr Asn Gly 20 25 30 Asp Cys Arg Val Glu Glu Cys Ala Leu Gly Gln Asp Leu Cys Arg Thr 35 40 45 Thr Ile Val Arg Leu Trp Glu Glu Gly Glu Glu Leu Glu Leu Val Glu 50 55 60 Lys Ser Cys Thr His Ser Glu Lys Thr Asn Arg Thr Leu Ser Tyr Arg 65 70 75 80 Thr Gly Leu Lys Ile Thr Ser Leu Thr Glu Val Val Gly Gly Gly Ser 85 90 95 Phe His Asn Asn Asp Thr Phe His Phe Leu Lys Cys Cys Asn Thr Thr 100 105 110 Lys Cys Asn Glu Gly Pro Ile Leu Glu Leu Glu Asn Leu Pro Gln Asn 115 120 125 Gly Arg Gln Cys Tyr Ser Cys Lys Gly Asn Ser Thr His Gly Cys Ser 130 135 140 Ser Glu Glu Thr Phe Leu Ile Asp Cys Arg Gly Pro Met Asn Gln Cys 145 150 155 160 Leu Val Ala Thr Gly Thr His Glu Pro Lys Asn Gln Ser Tyr Met Val 165 170 175 Arg Gly Cys Ala Thr Ala Ser Met Cys Gln His Ala His Leu Gly Asp 180 185 190 Ala Phe Ser Met Asn His Ile Asp Val Ser Cys Cys Thr Lys Ser Gly 195 200 205 Cys Asn His Pro Asp Leu Asp Val Gln Tyr Arg Ser Gly Ala Ala Pro 210 215 220 Gln Pro Gly Pro Ala His Leu Ser Leu Thr Ile Thr Leu Leu Met Thr 225 230 235 240 Ala Arg Leu Trp Gly Gly Thr Leu Leu Trp Thr 245 250 <210> 82 <211> 756 <212> DNA <213> Artificial sequence <400> 82 atgggtcacc cgccgctgct gccgctgctg ctgctgctcc acacctgcgt cccagcctct 60 Met - Gly - Thr - Pro - Arg - Ala - Ala - Ala - Ala - Ala - Ala - Ser - His - Thr - Cys - Val - Pro - Ser - Leu tggggcctgc ggtgcatgca gtgtaagacc aacggggatt gccgtgtgga agagtgcgcc 120 Trp - Gly - Leu - Arg - Val - His - Gln - Val - Lys - Thr - Asn - Gly - Ile - Ala - Val - Trp - Lys - Ser - Cys - Ala ctgggacagg acctctgcag gaccacgatc gtgcgcttgt gggaagaagg agaagagctg 180 Leu - Gly - Thr - Asp - Pro - Leu - Gln - Asp - His - Asp - Ser - Cys - Ala - Leu - Val - Glu - Lys - Glu - Glu - Glu - Leu gagctggtgg agaaaagctg tacccactca gagaagacca acaggaccct gagctatcgg 240 Glu - Leu - Val - Glu - Lys - Leu - Tyr - Pro - Thr - Gln - Glu - Lys - Thr - Asn - Arg - Pro - Glu - Leu - Ile - Gly actggcttga agatcaccag ccttaccgag gttgtgggag gaggatcatt ccacaacaac 300 Thr - Gly - Leu - Glu - Asp - Ile - Thr - Ser - Leu - Thr - Glu - Val - Val - Gly - Glu - Glu - Ile - Ile - Pro - His - Asn - Asn gacaccttcc acttcctgaa atgctgcaac accaccaaat gcaacgaggg cccaatcctg 360 Asp - Thr - Phe - His - Phe - Pro - Glu - Met - Leu - Gln - Thr - Thr - Thr - Asn - Ala - Asn - Glu - Gly - Pro - Asn - Leu gagcttgaaa atctgccgca gaatggccgc cagtgttaca gctgcaaggg gaacagcacc 420 Glu - Leu - Glu - Asn - Leu - Pro - Gln - Asn - Gly - Ala - Gln - Val - Thr - Ala - Gln - Gly - Asn - Ser - Thr catggatgct cctctgaaga gactttcctc attgactgcc gaggccccat gaatcaatgt 480 His - Trp - Asp - Ala - Pro - Leu - Glu - Glu - Thr - Phe - Pro - Ser - Asp - Cys - Ala - Glu - Ala - Pro - His - Asn - Ile - Val ctggtagcca ccggcactca cgaaccgaaa aaccaaagct atatggtaag aggctgtgca 540 Leu - Val - Ala - Thr - Gly - Thr - Thr - Glu - Pro - Lys - Thr - Lys - Ala - Tyr - Trp - Lys - Arg - Ala - Cys - Ala accgcctcaa tgtgccaaca tgcccacctg ggtgacgcct tcagcatgaa ccacattgat 600 Thr - Ala - Pro - Asn - Cys - Ala - Asn - Met - Pro - Thr - Gly - Asp - Ala - Phe - Ser - Met - Thr - His - Ile - Asp gtctcctgct gtactaaaag tggctgtaac cacccagacc tggatgtcca gtaccgcagt 660 Val - Ser - Pro - Ala - Val - Tyr - Lys - Ser - Gly - Cys - Asn - Thr - Pro - Asp - Trp - Asp - Val - Ser - Tyr - Arg - Ser ggggctgctc ctcagcctgg ccctgcccat ctcagcctca ccatcaccct gctaatgact 720 Gly - Ala - Ala - Ser - Ser - Ala - Trp - Ala - Leu - Pro - Ile - Ser - Ala - Ser - His - Ile - Thr - Leu - Asn - Asp gccagactgt ggggaggcac tctcctctgg acctaa 756 <210> 83 <211> 255 <212> PRT <213> Artificial sequence <400> 83 Met Gly Leu Lys Ile Thr Ser Leu Thr Glu Val Val Cys Gly Leu Asp 1 5 10 15 Leu Cys Asn Gln Gly Asn Ser Gly Arg Ala Val Thr Tyr Ser Arg Ser 20 25 30 Arg Tyr Leu Glu Cys Ile Ser Cys Gly Ser Ser Asp Met Ser Cys Glu 35 40 45 Arg Gly Arg His Gln Ser Leu Gln Cys Arg Ser Pro Glu Glu Gln Cys 50 55 60 Leu Asp Val Val Thr His Trp Ile Gln Glu Gly Glu Glu Gly Arg Pro 65 70 75 80 Lys Asp Asp Arg His Leu Arg Gly Cys Gly Tyr Leu Pro Gly Cys Pro 85 90 95 Gly Ser Asn Gly Phe His Asn Asn Asp Thr Phe His Phe Leu Lys Cys 100 105 110 Cys Asn Thr Thr Lys Cys Asn Glu Gly Pro Ile Leu Glu Leu Glu Asn 115 120 125 Leu Pro Gln Asn Gly Arg Gln Cys Tyr Ser Cys Lys Gly Asn Ser Thr 130 135 140 His Gly Cys Ser Ser Glu Glu Thr Phe Leu Ile Asp Cys Arg Gly Pro 145 150 155 160 Met Asn Gln Cys Leu Val Ala Thr Gly Thr His Glu Pro Lys Asn Gln 165 170 175 Ser Tyr Met Val Arg Gly Cys Ala Thr Ala Ser Met Cys Gln His Ala 180 185 190 His Leu Gly Asp Ala Phe Ser Met Asn His Ile Asp Val Ser Cys Cys 195 200 205 Thr Lys Ser Gly Cys Asn His Pro Asp Leu Asp Val Gln Tyr Arg Ser 210 215 220 Gly Ala Ala Pro Gln Pro Gly Pro Ala His Leu Ser Leu Thr Ile Thr 225 230 235 240 Leu Leu Met Thr Ala Arg Leu Trp Gly Gly Thr Leu Leu Trp Thr 245 250 255 <210> 84 <211> 768 <212> DNA <213> Artificial sequence <400> 84 atgggcttga agatcaccag ccttaccgag gttgtgtgtg ggttagactt gtgcaaccag 60 ggcaactctg gccgggctgt cacctattcc cgaagccgtt acctcgaatg catttcctgt 120 ggctcatcag acatgagctg tgagaggggc cggcaccaga gcctgcagtg ccgcagccct 180 gaagaacagt gcctggatgt ggtgacccac tggatccagg aaggtgaaga agggcgtcca 240 aaggatgacc gccacctccg tggctgtggc taccttcccg gctgcccggg ctccaatggt 300 ttccacaaca acgacacctt ccacttcctg aaatgctgca acaccaccaa atgcaacgag 360 ggcccaatcc tggagcttga aaatctgccg cagaatggcc gccagtgtta cagctgcaag 420 gggaacagca cccatggatg ctcctctgaa gagactttcc tcattgactg ccgaggcccc 480 atgaatcaat gtctggtagc caccggcact cacgaaccga aaaaccaaag ctatatggta 540 agaggctgtg caaccgcctc aatgtgccaa catgcccacc tgggtgacgc cttcagcatg 600 aaccacattg atgtctcctg ctgtactaaa agtggctgta accacccaga cctggatgtc 660 cagtaccgca gtggggctgc tcctcagcct ggccctgccc atctcagcct caccatcacc 720 ctgctaatga ctgccagact gtggggaggc actctcctct ggacctaa 768 <210> 85 <211> 150 <212> PRT <213> Artificial sequence <400> 85 Met Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala Thr Met 1 5 10 15 Pro Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp 20 25 30 Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln 35 40 45 Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile 50 55 60 Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg 65 70 75 80 Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro Phe Ser 85 90 95 Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp 100 105 110 Gln Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val Asn Gln 115 120 125 Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile 130 135 140 Ile Gln Asp Cys Met Trp 145 150 <210> 86 <211> 453 <212> DNA <213> Artificial sequence <400> 86 atggattttt ttcgggtagt ggaaaaccag cagcctcccg cgacgatgcc cctcaacgtt 60 agcttcacca acaggaacta tgacctcgac tacgactcgg tgcagccgta tttctactgc 120 gacgaggagg agaacttcta ccagcagcag cagcagagcg agctgcagcc cccggcgccc 180 agcgaggata tctggaagaa attcgagctg ctgcccaccc cgcccctgtc ccctagccgc 240 cgctccgggc tctgctcgcc ctcctacgtt gcggtcacac ccttctccct tcggggagac 300 aacgacggcg gtggcgggag cttctccacg gccgaccagc tggagatggt gaccgagctg 360 ctgggaggag acatggtgaa ccagagtttc atctgcgacc cggacgacga gaccttcatc 420 aaaaacatca tcatccagga ctgtatgtgg taa 453 <210> 87 <211> 215 <212> PRT <213> Artificial sequence <400> 87 Met Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala Thr Met 1 5 10 15 Pro Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp 20 25 30 Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln 35 40 45 Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile 50 55 60 Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg 65 70 75 80 Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro Phe Ser 85 90 95 Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp 100 105 110 Gln Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val Asn Gln 115 120 125 Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile 130 135 140 Ile Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys Leu Val 145 150 155 160 Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser 165 170 175 Pro Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser Leu Tyr 180 185 190 Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro Ser Val 195 200 205 Val Phe Pro Tyr Pro Leu Asn 210 215 <210> 88 <211> 648 <212> DNA <213> Artificial sequence <400> 88 atggattttt ttcgggtagt ggaaaaccag cagcctcccg cgacgatgcc cctcaacgtt 60 agcttcacca acaggaacta tgacctcgac tacgactcgg tgcagccgta tttctactgc 120 gacgaggagg agaacttcta ccagcagcag cagcagagcg agctgcagcc cccggcgccc 180 agcgaggata tctggaagaa attcgagctg ctgcccaccc cgcccctgtc ccctagccgc 240 cgctccgggc tctgctcgcc ctcctacgtt gcggtcacac ccttctccct tcggggagac 300 aacgacggcg gtggcgggag cttctccacg gccgaccagc tggagatggt gaccgagctg 360 ctgggaggag acatggtgaa ccagagtttc atctgcgacc cggacgacga gaccttcatc 420 aaaaacatca tcatccagga ctgtatgtgg agcggcttct cggccgccgc caagctcgtc 480 tcagagaagc tggcctccta ccaggctgcg cgcaaagaca gcggcagccc gaaccccgcc 540 cgcggccaca gcgtctgctc cacctccagc ttgtacctgc aggatctgag cgccgccgcc 600 tcagagtgca tcgacccctc ggtggtcttc ccctaccctc tcaactaa 648 <210> 89 <211> 1706 <212> DNA <213> Artificial sequence <400> 89 aagcttgcat gcctgcaggt caacatggtg gagcacgaca cacttgtcta ctccaaaaat 60 atcaaagata cagtctcaga agaccaaagg gcaattgaga cttttcaaca aagggtaata 120 tccggaaacc tcctcggatt ccattgccca gctatctgtc actttattgt gaagatagtg 180 gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc catcgttgaa 240 gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa 300 aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgataa catggtggag 360 cacgacacac ttgtctactc caaaaatatc aaagatacag tctcagaaga ccaaagggca 420 attgagactt ttcaacaaag ggtaatatcc ggaaacctcc tcggattcca ttgcccagct 480 atctgtcact ttattgtgaa gatagtggaa aaggaaggtg gctcctacaa atgccatcat 540 tgcgataaag gaaaggccat cgttgaagat gcctctgccg acagtggtcc caaagatgga 600 cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa 660 gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca ctatccttcg 720 caagaccctt cctctatata aggaagttca tttcatttgg agaggacctc gacctcaaca 780 caacatatac aaaacaaacg aatctcaagc aatcaagcat tctacttcta ttgcagcaat 840 ttaaatcatt tcttttaaag caaaagcaat tttctgaaaa ttttcaccat ttacgaacga 900 tatctagagg atccttaatt aacatggtga gcaagggcga ggagctgttc accggggtgg 960 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 1020 t gcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 1020 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 1080 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 1080 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg cagtgcttcg 1140 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg cagtgcttcg 1140 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 1200 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 1200 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 1260 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 1260 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1320 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1320 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1380 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1380 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 1440 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 1440 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 1500 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 1500 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca 1560 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca 1560 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 1620 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 1620 gcatggacga gctgtacaag ggtaccggcg cgcctgagca gaaactcatc tctgaagagg 1680 gcatggacga gctgtacaag ggtaccggcg cgcctgagca gaaactcatc tctgaagagg 1680 atctgtagtg agagctcgaa tttccc 1706

Claims

1. Use in any one of P1 - P4 as follows: a), a plant JAZ protein; b), a biological material related to the plant JAZ protein described in a); the related biological material is any one of the following: 1) a nucleic acid molecule encoding the plant JAZ protein; 2) an expression cassette, expression vector or recombinant microorganism containing the nucleic acid molecule; P1, Preparation of a product for treating and / or preventing tumors in humans or animals; P2, Preparation of a product for inhibiting the occurrence and / or development of tumors in humans or animals; P3, Preparation of a product for inhibiting the proliferation of tumor cells in humans or animals; P4, Preparation of a product for promoting apoptosis of tumor cells in humans or animals; The plant JAZ protein is any one of the following: (A1) A protein with an amino acid sequence shown in SEQ ID No. 4; (A2) A fusion protein obtained by linking a protein tag to the N - terminus and / or C - terminus of the protein defined in (A1); the protein tag is a Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag and / or SUMO tag; The human or animal tumor is selected from any one of the following: ovarian cancer, breast cancer, cervical cancer, lung cancer; The human or animal tumor cells are selected from any one of the following: ovarian cancer cells, breast cancer cells, cervical cancer cells, lung cancer cells.

2. The application according to claim 1, wherein: The nucleic acid molecule encoding the plant JAZ protein is the DNA molecule shown in SEQID No.

39.

3. The application according to claim 1 or 2, characterized in that: The ovarian cancer cells are SKOV3 cells; the breast cancer cells are MCF7 cells; the cervical cancer cells are Hela cells; the lung cancer cells are A549 cells.

4. The application according to claim 1 or 2, characterized in that: The product is a drug.

5. The application according to claim 4, characterized in that: The drug is prepared by adding pharmaceutically acceptable excipients on the basis of a) or b) described in claim 1.

6. The application according to claim 4, wherein: The drug is a tablet, pill, granule, capsule, injection or oral liquid.

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