Stable type III interferon protein and its fusion protein

Abstract

The present application provides a stable type III interferon protein and its fusion protein, conjugate, pharmaceutical composition, product, and kit. The stable type III interferon protein provided herein has good activity and stability and can effectively prevent and / or treat viral infectious diseases, such as diseases caused by respiratory syncytial virus.

Description

Technical Field

[0001] The present application relates to type III interferon proteins and their fusion proteins, their conjugates, and pharmaceutical compositions containing them that can be used to prevent and treat diseases caused by pathogenic infections in human and animal individuals, and in particular, type III interferon proteins and their fusion proteins that can be used to prevent and treat viral infectious diseases, such as the prevention and treatment of diseases caused by respiratory syncytial virus (RSV). Background Art

[0002] Respiratory syncytial virus (RSV) is an enveloped, non-segmented, single-stranded, negative-sense RNA virus belonging to the Pneumoviridae family and the genus Orthopneumovirus. Human RSV has only one serotype, which is divided into two major subtypes based on antigenicity: type A and type B. These subtypes differ significantly in their major structural proteins. First discovered in children in 1956, RSV primarily causes respiratory tract infections and has become a leading cause of respiratory hospitalization in infants and children under five years of age. It is also a major pathogen causing severe respiratory illness in adults, the elderly, and immunocompromised patients.

[0003] Prevention and treatment of the virus are generally carried out using vaccines and neutralizing antibodies. However, there is currently no safe and effective RSV vaccine or treatment available clinically. Early formalin-inactivated vaccines not only failed to provide adequate protection to recipients, but also caused enhanced RSV disease (ERD) and even infant mortality. Currently, the only approved treatment is ribavirin, but due to its complex administration and significant side effects, it is only used in high-risk children. Although a human monoclonal antibody, Palivizumab, has been approved for the prevention of RSV infection and treatment to reduce the severity of the disease, it is difficult to use widely due to the high cost of monthly injections. Currently, there is no effective and safe anti-RSV vaccine or specific antiviral drug, making the development of new and effective antiviral drugs urgent.

[0004] Interferon (IFN), as one of the main players in the body's innate antiviral immune response, has attracted much attention in the pathogenesis of respiratory infections. Type I and type II IFNs are considered to be representative cytokines in the early host defense mechanism against viral infections. The so-called "type I" interferons include interferon α, interferon β, interferon ω, interferon δ, and interferon τ. Currently, interferon γ is the only type II interferon. Type III interferon, or IFN-λ, is a recently discovered family of cytokines that includes interferons λ1, λ2, and λ3, also known as IL-29, IL-28A, and IL-28B.

[0005] Type III IFNs have similar antiviral properties to type I IFNs, but because their receptors are tissue-specific, their expression is restricted to epithelial cells. This allows them to exert their protective effects primarily on the epithelial surface, making their antiviral effects more targeted and less prone to systemic side effects than type I and type II IFNs. IFN-λ consists of three subtypes: IL-29 (IFN-λ1), IL-28A (IFN-λ2), and IL-28B (IFN-λ3). The order of antiviral activity is IFN-λ3 > IFN-λ1 > IFN-λ2, with IFN-λ1 being highly expressed in respiratory epithelial cells. Therefore, IFN-λ1 holds promise as a targeted antiviral drug with minimal side effects for the treatment of respiratory viral infections.

[0006] Some progress has been made in exploring the antiviral effects of type III interferons and their relationship to respiratory viruses. Type III interferons can inhibit the replication of viruses such as influenza virus and hepatitis C virus, and have recently been shown to effectively inhibit the replication of the novel coronavirus. Furthermore, type III interferons can indirectly inhibit viruses by upregulating the expression of MHC class I molecules. Studies have shown that mucosal epithelial cells in mice with type III interferon receptor knockout significantly increase viral susceptibility, while mucosal epithelial cells in mice with type I interferon receptor knockout do not exhibit such changes. Furthermore, some researchers have found that exogenous IFN-λ can cure persistent and recurrent viral infections, whereas IFN-α has no such effect. Research on IFN-λ has shown strong antiviral activity in experimental animals, in vitro studies, and clinical studies, and even plays a dominant role in respiratory viral infections. In clinical studies against hepatitis C virus infection, the antiviral efficacy of IFN-λ is comparable to that of type I interferons.

[0007] However, as protein drugs, interferons (type I, II, and III) are subject to significant limitations in clinical use due to factors such as poor stability, low activity, and a short half-life in vivo. Therefore, developing IFN-λ1 into a stable, highly active, safe, and effective RSV vaccine or therapeutic would be a significant boon for RSV infection.

[0008] Since type III interferon proteins are mainly produced by Escherichia coli, the N-terminal M is easily oxidized and has low bioavailability. Therefore, there is an urgent need to further develop type III interferon proteins or their fusion proteins with better stability, higher activity, better affinity and better in vivo efficacy. Summary of the Invention

[0009] In summary, in order to solve the problems of the prior art, the present application provides a type III interferon protein with better stability, higher activity, fewer adverse reactions and can be used for aerosol inhalation therapy. On the one hand, the present application provides a type III interferon protein, comprising a substitution mutation of the 161st or 162nd amino acid on the amino acid sequence shown in SEQ ID NO.1, wherein the aspartic acid (D) at position 161 or the glycine (G) at position 162 is replaced by other natural amino acids. In some embodiments, the type III interferon protein of the present application comprises aspartic acid (D) at position 161 on the amino acid sequence shown in SEQ ID NO.1 replaced by glutamic acid, threonine or serine, or glycine (G) at position 162 is replaced by an aliphatic amino acid. In some embodiments, the type III interferon protein of the present application further comprises a substitution mutation of the 165th amino acid from cysteine (C) to serine (S) on the amino acid sequence shown in SEQ ID NO.1. In the present application, the amino acid sequence of the full-length wild-type type III interferon protein including the signal peptide is shown in SEQ ID NO.3, which consists of 200 amino acids, of which amino acids 1-19 are signal peptides and amino acids 20-200 (181aa) constitute the mature protein of type III interferon (amino acid sequence as shown in SEQ ID NO.2). The type III interferon protein of the present application is designed starting from position 26 starting from the N-terminus of the wild-type protein shown in SEQ ID NO.3. The type III interferon protein of the present application comprises an amino acid substitution mutation at position 161 or 162 of the amino acid sequence shown in SEQ ID NO.1.

[0010] In some embodiments of the present application, the present application provides a type III interferon protein, comprising a substitution mutation of amino acid 161 or 162 on the amino acid sequence shown in SEQ ID NO.1, wherein the aspartic acid (D) at position 161 or the glycine (G) at position 162 is substituted by other natural amino acids, for example, by an amino acid selected from the following: glycine, alanine, valine, leucine, isoleucine, methionine (methionine), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine or histidine.

[0011] Those skilled in the art will understand that positions 161 and 162 of SEQ ID NO.1 correspond to the corresponding positions of SEQ ID NO.2 (mature protein of wild-type type III interferon) and SEQ ID NO.3 (full-length protein of wild-type type III interferon including a signal peptide). Therefore, amino acid mutations at the corresponding positions of SEQ ID NO.2 or SEQ ID NO.3 (or amino acid sequences of different lengths derived from SEQ ID NO.2 or SEQ ID NO.3) are also covered by the protection scope of this application. The protection scope of this application also covers type III interferon proteins containing substitution mutations at the corresponding positions of positions 161 and 162 derived from SEQ ID NO.2 or SEQ ID NO.3 but having amino acid sequences of different lengths.

[0012] In some embodiments of the above-mentioned type III interferon protein, the type III interferon protein comprises a substitution mutation of amino acid 161 or 162 on the amino acid sequence shown in SEQ ID NO. 1, wherein the aspartic acid (D) at position 161 or the glycine (G) at position 162 is substituted with other natural amino acids, and further comprises an initial methionine (M). This is because when the type III interferon protein is expressed in prokaryotic cells (such as E. coli), the expressed type III interferon protein has a methionine at the N-terminus or amino terminus.

[0013] In some embodiments of the present application, the type III interferon protein of the present application comprises a substitution mutation of the amino acid at position 161 or 162 on the amino acid sequence shown in SEQ ID NO.1, that is, a mutation at position 161 or 162, starting from the N-terminus or amino-terminus of the protein shown in SEQ ID NO.1, for example, the aspartic acid (D) at position 161 or the glycine (G) at position 162 is replaced by other natural amino acids. In some embodiments, the type III interferon protein of the present application is a mutant protein expressed in prokaryotic cells (such as E. coli) and comprises a substitution mutation at position 162 or 163 (because of the addition of the N-terminal M, starting from M), for example, the aspartic acid (D) at position 162 or the glycine (G) at position 163 is replaced by other natural amino acids. In a specific embodiment, the type III interferon protein of the present application is as shown in SEQ ID NO.4-9.

[0014] In some embodiments of the above-mentioned type III interferon protein, the type III interferon protein comprises a substitution mutation of the amino acid at position 161 or 162 on the amino acid sequence shown in SEQ ID NO.1, for example, wherein the aspartic acid (D) at position 161 is substituted with glutamic acid, threonine or serine, or the glycine (G) at position 162 is substituted with an aliphatic amino acid. In some embodiments, the type III interferon protein of the present application is a mutant protein expressed in a prokaryotic cell (e.g., E. coli), comprising a substitution mutation at position 162 or 163 (because of the addition of the N-terminal M, counting from M), for example, wherein the aspartic acid (D) at position 162 is substituted with glutamic acid, threonine or serine, or the glycine (G) at position 163 is substituted with an aliphatic amino acid.

[0015] In some embodiments of the above-mentioned type III interferon protein, the type III interferon protein comprises or consists of the following amino acid sequence: SEQ ID NO.4, SEQ ID NO.6 or SEQ ID NO.8.

[0016] In some embodiments of the above-mentioned type III interferon protein, the type III interferon protein further comprises a substitution mutation from cysteine (C) to serine (S) at position 165 of the amino acid sequence shown in SEQ ID NO. 1. As described above, when the type III interferon protein is expressed in prokaryotic cells (e.g., E. coli), the substitution mutation from cysteine (C) at position 165 to serine (S) will appear at position 166.

[0017] In some embodiments of the above-mentioned type III interferon protein, the type III interferon protein comprises or consists of the following amino acid sequence: SEQ ID NO.5, SEQ ID NO.7 or SEQ ID NO.9.

[0018] In some embodiments of the present application, the present application provides a type III interferon protein (IL29 DE) as shown in SEQ ID NO.4; in some embodiments, the present application provides a type III interferon protein (IL29 DE + CS) as shown in the following SEQ ID NO.5; in some embodiments, the present application provides a type III interferon protein (IL29 DS) as shown in the following SEQ ID NO.6; in some embodiments, the present application provides a type III interferon protein (IL29 DS + CS) as shown in the following SEQ ID NO.7; in some embodiments, the present application provides a type III interferon protein (IL29 GA) as shown in the following SEQ ID NO.8; in some embodiments, the present application provides a type III interferon protein (IL29 GA + CS) as shown in SEQ ID NO.9; in some embodiments of the above-mentioned type III interferon protein, the type III interferon protein further comprises a short sequence to promote protein purification (e.g., a short sequence of 6 histidines), or a short amino acid sequence to prolong half-life.

[0019] Type III interferon proteins such as those shown in SEQ ID NOs. 4, 5, 8, and 9 are relatively more stable and have higher specific activity than wild-type proteins. They are suitable for use in aerosol inhalation therapy to prevent and / or treat respiratory diseases, where a nebulizer is required to atomize a drug solution into tiny particles for inhalation into the respiratory tract and lungs, where the drug is deposited. Their excellent results are documented in Chinese patent application number CN 202110169885.8.

[0020] The present application, on the other hand, provides a stable type III interferon protein, whose structure is the sequence after the N-terminal M of the IL-29DE+CS protein is removed and a stabilizing amino acid is added. For example, M is removed and the added stabilizing amino acid is glycine (G), alanine (A), proline (P), or serine (S). Preferably, the added stabilizing amino acid is glycine (G).

[0021] In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 10. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 11. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 12. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 13.

[0022] The present application, on the other hand, provides a stable type III interferon protein, whose structure is the sequence after the N-terminal M of the IL-29DE protein is removed and a stabilizing amino acid is added. For example, M is removed and the added stabilizing amino acid is glycine (G), alanine (A), proline (P), or serine (S). Preferably, the added stabilizing amino acid is glycine (G).

[0023] In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 35. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 36. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 37. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 38.

[0024] The present application, on the other hand, provides a stable type III interferon protein having a structure in which the N-terminal M of the IL-29GA protein is removed and a stabilizing amino acid is added. For example, M is removed and the stabilizing amino acid added is glycine (G), alanine (A), proline (P), or serine (S). Preferably, the stabilizing amino acid added is glycine (G).

[0025] In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 39. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 40. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 41. In some embodiments, the present application provides a stable type III interferon protein as shown in SEQ ID NO. 42.

[0026] The present application, on the other hand, provides a stable type III interferon protein having a structure such that the N-terminal M of the IL-29GA+CS protein is removed and a stabilizing amino acid is added. For example, M is removed and the stabilizing amino acid added is glycine (G), alanine (A), proline (P), or serine (S). Preferably, the stabilizing amino acid added is glycine (G).

[0027] In some embodiments, the present application provides a stabilized type III interferon protein as shown in SEQ ID NO. 43. In some embodiments, the present application provides a stabilized type III interferon protein as shown in SEQ ID NO. 44. In some embodiments, the present application provides a stabilized type III interferon protein as shown in SEQ ID NO. 45. In some embodiments, the present application provides a stabilized type III interferon protein as shown in SEQ ID NO. 46.

[0028] The present application, on the other hand, provides a fusion protein of a stable type III interferon protein, which comprises the stable type III interferon protein and further comprises a linker and / or a binding sequence targeting epithelial cells. In some embodiments, the present application relates to a fusion protein, wherein the binding sequence targeting epithelial cells is located at the N-terminus or C-terminus of the stable type III interferon protein, preferably at the C-terminus. In some embodiments, the present application relates to a fusion protein, wherein the linker is located at the N-terminus or C-terminus of the stable type III interferon protein, preferably at the C-terminus. In some embodiments, the present application relates to a fusion protein, wherein its structure is A-Ln-B, wherein A is a stable type III interferon protein, L is a linker, B is a binding sequence targeting epithelial cells, and n is 0 to 6. A is the stable type III interferon protein mentioned above.

[0029] The linker is a glycine-rich peptide, or a peptide having GGGGS (SEQ ID NO.14), wherein n is 0, 1, 2, 3, 4, 5 or 6. Preferably, the linker is added to prevent potential undesirable domain interactions and optimize the in vivo function and stability of the stable type III interferon protein of the present invention. Although these linkers can be of any length and composed of any amino acid combination, their length preferably does not exceed the length required to prevent undesirable domain interactions and / or optimize biological function and / or stability. The linker is preferably rich in serine-glycine, and its length preferably does not exceed 30 amino acids. The length of the linker is more preferably no more than 20 amino acids, and most preferably no more than 15 amino acids. The preferred linker contains a repeat of the sequence GGGGS, more preferably GGGGS.

[0030] B represents a binding sequence that targets epithelial cells, particularly those in the lungs. For example, heparin and heparan sulfate are common glycosaminoglycans (GAGs) ubiquitously present on cell membranes, particularly on the surface of respiratory epithelial cells. Currently, many heparin / heparan sulfate-binding proteins have been reported, including human amphiregulin, members of the FGF family, and AAMPs, all of which exhibit extremely high affinity for heparin (nM).

[0031] The binding sequence targeting epithelial cells is, for example, the sequence shown in SEQ ID NO.15, SEQ ID NO.16, SEQ ID NO.17, and SEQ ID NO.18, preferably SEQ ID NO.15 and SEQ ID NO.17.

[0032] In some embodiments, the present application relates to a stable type III interferon protein or a fusion protein thereof, wherein the protein is fused to another polypeptide or protein at the N-terminus or C-terminus. For example, it can be fused with human albumin, transferrin, the Fc portion of a human IgG molecule, etc. to form a fusion protein to increase the half-life of the protein in vivo. The stable type III interferon protein or its fusion protein can also be fused with other proteins targeting different targets to achieve a combined preventive and / or therapeutic effect, thereby improving the preventive and / or therapeutic effect of the drug. For example, it can be fused with DAS181 to enhance the activity of the mutant protein of the present invention in preventing and / or treating viruses. DAS181 uses a unique host-directed approach to block respiratory viral infection by disconnecting sialic acid receptors in the human respiratory tract. These receptors bind to most major respiratory viruses, leading to patient infection. DAS181 has shown antiviral activity against four major respiratory viruses, including influenza virus (IFV), parainfluenza virus (PIV), metapneumovirus (MPV), and human enterovirus-68 (EV-68).

[0033] In some embodiments, the present application relates to a conjugate of a stable type III interferon protein or its fusion protein, wherein the stable type III interferon protein or its fusion protein is coupled to a polyalkoxy compound. The polyalkoxy compound is, for example, polyethylene glycol (PEG), such as a linear or branched polyethylene glycol, specifically monomethoxy polyethylene glycol propionaldehyde (mPEG propionaldehyde), such as 20kD, 30kD or 40kD mPEG propionaldehyde. The process of modifying the stable type III interferon protein or its fusion protein with PEG to form a conjugate is called PEGylation. The PEGylation of the stable type III interferon protein or its fusion protein in the present application can be carried out by any of the PEGylation methods known in the art, such as using an acylation reaction or an alkylation reaction. The therapeutic half-life of the protein can be artificially increased by coupling one or more PEGs that increase the overall size to the protein to avoid rapid degradation in the body.

[0034] In some embodiments, the present application relates to a polynucleotide encoding any of the above-mentioned stabilized type III interferon proteins or a fusion protein thereof.

[0035] In some embodiments, the present application relates to a vector comprising the above-mentioned polynucleotide. Briefly, a polynucleotide encoding a stable type III interferon protein or a fusion protein thereof is inserted into a suitable expression vector such that the polynucleotide is operably linked to a multiple cloning site to express the corresponding protein.

[0036] The vector can be a pET series vector such as pET-3a, pET-9a, pET-11a, pET-14b, pET-15b, pET-16b, pET-17b, pET-19b, pET-20b, pET-21a, pET-22b, pET-23a(+), pET-24a, pET-25b(+), pET-26b(+), pET-27b(+), pET-28a(+), pET-29a(+), pET-30a(+), pET-31b(+), pET-32a( +), pET-39b(+), pET-40b(+), pET-41a(+), pET-42a(+), pET-43.1a(+), pET-44b, pET-45b, pET-47b, pET-48b, pET-49b(+), pET-50b(+), pET-51b(+), pET-52b(+); pMal series vectors such as pMal-c2X and pMal-c5X; pGEX series vectors such as pGEX-6p-1 and pGEX-6P-2; pRSET series vectors such as pRSET A, B and C; pTricHis series vectors such as pTrcHis A, B and C. Preferably, the vector is pET-30a(+).

[0037] In some embodiments, the present application relates to a host cell comprising the above-mentioned polynucleotide or vector. The host cell is used to express the stable type III interferon protein or its fusion protein described herein. A host cell refers to a recipient cell that accepts an exogenous gene by, for example, transformation or transduction. Common host cells include prokaryotic recipient cells and eukaryotic recipient cells. The present application prefers prokaryotic recipient cells as its host cells. Among them, the host cell is an Escherichia coli competent cell BL21 (DE3), Tuner (DE3), Origami (DE3), Rosetta (DE3), JM109 (DE3), BL21 star (DE3), Rosetta-gami (DE3), Rosetta-gami B (DE3), BL21 (DE3) pLysS or BL21 star (DE3) pLysS. Preferably, the host cell is an Escherichia coli competent cell BL21 (DE3).

[0038] In some embodiments, the present application relates to a pharmaceutical composition comprising the aforementioned stabilized type III interferon protein, or its fusion protein, or their conjugate and a pharmaceutically acceptable carrier.

[0039] The pharmaceutical composition may be in the form of an injection, tablet, capsule, inhaler, suppository, etc. Preferably, the pharmaceutical composition is an inhaler, such as a dry powder inhaler or a liquid inhaler, such as a nebulizer inhaler, aerosol, soft mist, and spray, and is administered through an inhalation device, such as a nebulizer inhaler, a metered dose inhaler, or a dry powder inhaler.

[0040] In some embodiments, the pharmaceutical composition comprising a stabilized type III interferon protein, or a fusion protein or conjugate thereof, further comprises a carrier suitable for pulmonary administration for the prevention and / or treatment of diseases caused by various respiratory syncytial viruses.

[0041] In some embodiments, the present application relates to a method for preparing a stable type III interferon protein or a fusion protein thereof, comprising: introducing a nucleic acid (polynucleotide) encoding the above-mentioned stable type III interferon protein or its fusion protein into a host cell, so that the host cell expresses the stable type III interferon protein or its fusion protein, for example, inserting the nucleic acid into an expression vector; transferring the vector into a host cell (such as Escherichia coli) to obtain a corresponding host cell (engineered bacteria) expressing the protein; culturing (such as fermentation) the host cell (engineered bacteria) to induce the host cell (engineered bacteria) to express the protein; and harvesting the protein.

[0042] In some embodiments, the above method further comprises purifying the harvested protein and / or renaturing the protein.

[0043] Beneficial effects

[0044] The stabilized type III interferon protein and its fusion protein of the present application have the following beneficial effects:

[0045] 1. Better stability

[0046] By replacing the second amino acid with glycine (G), methionyl aminopeptidase removes the N-terminal M, resulting in a G as the first amino acid. This significantly reduces N-terminal oxidation. When the N-terminus is M, the N-terminal oxidation rate is 2.5% after 5 days of exposure to light at 25°C. However, when the N-terminus is G, the N-terminal oxidation rate is zero under the same conditions. This significantly improves the stability of the sample, making it possible to store the drug at room temperature.

[0047] 2. Higher activity

[0048] When a heparin-binding sequence was added to the C-terminus of the stabilized type III interferon protein, the cell activity detected by the cell activity assay was significantly increased, especially when AR or FGF18 was added. 50 It decreased from 0.968nM to 0.138 and 0.185nM, a decrease of nearly 5 to 7 times.

[0049] 3. Pharmacokinetic effect is significantly improved

[0050] In vivo drug exposure AUC last and maximum blood concentration C max The results were at least 1.49 times, and even as high as 1.79 times, higher than those without the linker and heparin binding sequence, indicating a significant increase in drug exposure and maximum blood concentration in mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 This is the electrophoresis diagram of protein expression after adding heparin binding sequence.

[0052] Figure 2 The SPR results of each protein and heparin (Biacore 8K).

[0053] Figure 3 The cytological activities of M1 and the combination of each protein.

[0054] Figure 4 The pharmacokinetic results of each protein in mice are shown in Figure 2. DETAILED DESCRIPTION

[0055] The present application provides a stable type III interferon protein and its fusion protein, a method for preparing the stable type III interferon protein and its fusion protein, and the use of the stable type III interferon protein, the stable type III interferon protein fusion protein, their conjugates or pharmaceutical compositions comprising the stable type III interferon protein, the stable type III interferon protein fusion protein, their conjugates and a pharmaceutically acceptable carrier in preventing and / or treating viral infectious diseases, such as preventing and treating diseases caused by respiratory syncytial virus. The inventors screened multiple heparin binding sequences and linker insertion positions, and finally found that the stable type III interferon protein provided by the present application, after adding linker and heparin binding sequence to the C-terminus of the stable type III interferon protein, the cell activity of the obtained stable type III interferon protein was significantly increased, and the N-terminus was oxidized to 0, which greatly improved the stability of the sample, so that the drug can be placed at room temperature, has higher affinity, and the in vivo drug exposure AUClast and maximum blood concentration C max The results show that the N-terminal M of the existing antiviral protein drugs is easily oxidized, has low bioavailability, low activity, poor stability, poor affinity, and serious adverse reactions.

[0056] Common methods for detecting the activity of stable type III interferon proteins or their fusion proteins include cytopathic assays and reporter gene assays. The cytopathic assay involves using human retinal pigment epithelial (ARPE)-19 cells infected with vesicular stomatitis virus (VSV) and evaluating the degree of protection of the cells by the stable type III interferon protein using a crystal violet staining colorimetric method, thereby evaluating the activity of the stable type III interferon protein (Kotenko Sergei V, Gallagher Grant, Baurin Vitaliy V et al. IFN-lambdas mediate antiviral protection through a distinct class II cytokine receptor complex. [J]. Nat. Immunol., 2003, 4: 69-77).

[0057] The reporter gene method involves linking the interferon-stimulated response element (ISRE) promoter to luciferase cDNA and transfecting the cells into HEK293 cells. After stimulation with a stable type III interferon protein, the activity of the stable type III interferon protein is evaluated by measuring the luciferase enzyme in the supernatant of HEK293 blood cells (LaFleur DW, Nardelli B, Tsareva T et al. Interferon-kappa, a novel type I interferon expressed in human keratinocytes. [J]. J. Biol. Chem., 2001, 276: 39765-71.).

[0058] There are many methods for testing the stability of stabilized type III interferon proteins or their fusion proteins. For example, reversed-phase HPLC can be used to analyze impurities with different hydrophobicity and polarity; ion-exchange HPLC can be used to separate impurities with large charge differences; and molecular sieve exclusion chromatography can be used to analyze dimers, polymers, and monomers. Each test method focuses on different aspects, but all can be used to characterize protein purity and provide insights into protein stability.

[0059] As used herein, "type III interferon protein," i.e., IFN-λ1, refers to a mutant interleukin 29 (IL29) protein. As used herein, "stabilized type III interferon protein" refers to a protein expressed by removing the N-terminal amino acid sequence of the mutant interleukin 29 (IL29) protein and adding a stabilizing amino acid sequence. As used herein, "stabilized type III interferon protein fusion protein" refers to a protein in which the N-terminal amino acid sequence of the mutant interleukin 29 (IL29) protein is removed and a stabilizing amino acid sequence is added, followed by the addition of a linker and / or epithelial cell-targeting binding sequence to the N- or C-terminus.

[0060] As used herein, the epithelial cell targeting binding sequence is preferably a heparin binding sequence.

[0061] As used herein, the term "respiratory syncytial virus" refers to a negative-sense, single-stranded, enveloped RNA virus belonging to the family Paramyxoviridae. RSV is a member of the pneumovirus subfamily and occurs in both round and filamentous forms. It is a causative agent of human respiratory illness, primarily causing severe illness in infants and immunosuppressed patients, particularly lung transplant recipients. RSV is the most common pathogen of viral pneumonia in children under five years of age and a cause of sudden infant death. Currently, there is a lack of effective methods for preventing RSV. Intranasal administration of a temperature-sensitive viral vaccine to children can produce local secretory IgA antibodies and is currently under investigation. Treatment is primarily supportive and symptomatic, with antibiotics available for secondary bacterial infection. Children and some severely immunocompromised patients are given ribavirin, sometimes in combination with RSV immune globulin and / or palivizumab. However, with the widespread use of ribavirin, clinical observations have found that ribavirin resistance has led to a gradual decline in its clinical efficacy. In addition, adverse reactions such as leukopenia and rash have also occurred during its use.

[0062] In some embodiments of the present application, a stable type III interferon protein is provided, comprising an amino acid sequence selected from any one of SEQ ID NOs. 10-13, SEQ ID NOs. 35-38, SEQ ID NOs. 39-42, and SEQ ID NOs. 43-46.

[0063] In some embodiments of the present application, a fusion protein of a stable type III interferon protein is provided, which comprises the stable type III interferon protein described above and further comprises a linker and / or a binding sequence targeting epithelial cells. Its structure is A-Ln-B, wherein A is an amino acid sequence selected from any one of SEQ ID NO.10-13, SEQ ID NO.35-38, SEQ ID NO.39-42, and SEQ ID NO.43-46, L is a linker, B is a binding sequence targeting epithelial cells, and n is 0-6. The linker is a glycine-rich peptide, and the linker is a peptide having GGGGS (SEQ ID NO.14). The binding sequence targeting epithelial cells is a heparin binding sequence, preferably selected from any one of the following: SEQ ID NO.15-18, preferably the binding sequence targeting epithelial cells is a sequence shown in SEQ ID NO.15 and SEQ ID NO.17. The binding sequence targeting epithelial cells is located at the N-terminus or C-terminus of the stable type III interferon protein, preferably the C-terminus. The linker is located at the N-terminus or C-terminus of the stable type III interferon protein, preferably the C-terminus.

[0064] In some embodiments of the present application, a stable fusion protein of a type III interferon protein is provided, the amino acid sequence of which is selected from any one of SEQ ID NOs. 28 to 34, 47 to 49. In some embodiments of the present application, a stable fusion protein of a type III interferon protein is provided, wherein the fusion protein is fused to another polypeptide or protein at the N-terminus or C-terminus.

[0065] In some embodiments of the present application, a stable conjugate of a fusion protein of a type III interferon protein is provided, wherein the fusion protein is conjugated to a polyalkoxy compound, such as polyethylene glycol, such as a linear or branched polyethylene glycol, specifically monomethoxypolyethylene glycol propionaldehyde (mPEG propionaldehyde), such as 20kD, 30kD or 40kD mPEG propionaldehyde.

[0066] In some embodiments of the present application, a polynucleotide is provided, which encodes a stabilized type III interferon protein or a fusion protein of a stabilized type III interferon protein.

[0067] In some embodiments of the present application, a vector comprising the above-mentioned polynucleotide is provided.

[0068] In some embodiments of the present application, a host cell comprising the above polynucleotide or a vector comprising the above polynucleotide is provided.

[0069] In some embodiments of the present application, a method for producing a stable type III interferon protein or a fusion protein of a stable type III interferon protein is provided, which comprises causing the above-mentioned host cell to express the stable type III interferon protein or the fusion protein of the stable type III interferon protein, and isolating the stable type III interferon protein or the fusion protein of the stable type III interferon protein from the host cell culture.

[0070] In some embodiments of the present application, a pharmaceutical composition is provided, which comprises a stable type III interferon protein, a fusion protein of a stable type III interferon protein, a conjugate of a stable type III interferon protein and a fusion protein thereof, and a pharmaceutically acceptable carrier.

[0071] In some embodiments of the present application, a product or kit is provided, comprising a stable type III interferon protein, a fusion protein of a stable type III interferon protein, a conjugate of a stable type III interferon protein and its fusion protein, or a pharmaceutical composition comprising a stable type III interferon protein, a fusion protein of a stable type III interferon protein, a conjugate of a stable type III interferon protein and its fusion protein and a pharmaceutically acceptable carrier.

[0072] In some embodiments of the present application, a stable type III interferon protein, a fusion protein of a stable type III interferon protein, a conjugate of a stable type III interferon protein and its fusion protein, or a pharmaceutical composition comprising a stable type III interferon protein, a fusion protein of a stable type III interferon protein, a conjugate of a stable type III interferon protein and its fusion protein and a pharmaceutically acceptable carrier is provided for use in the preparation of a medicament for preventing and / or treating a viral infectious disease. The viral infectious disease is a disease caused by respiratory syncytial virus.

[0073] In some embodiments of the present application, a method for preventing and / or treating a viral infectious disease is provided, comprising administering to a subject a therapeutically effective amount of a stable type III interferon protein, a fusion protein of a stable type III interferon protein, a conjugate of a stable type III interferon protein and its fusion protein, or a pharmaceutical composition comprising a stable type III interferon protein, a fusion protein of a stable type III interferon protein, a conjugate of a stable type III interferon protein and its fusion protein, and a pharmaceutically acceptable carrier. The viral infectious disease is a disease caused by respiratory syncytial virus.

[0074] This is further illustrated by the following non-limiting examples.

[0075] Example

[0076] Example 1 Preparation of stable type III interferon protein

[0077] 1.1 Construction of recombinant plasmid

[0078] After chemical synthesis, the nucleotide sequence capable of expressing the amino acid sequence of the protein shown in Table 1 was synthesized, double-digested with NdeI and XhoI, constructed into the prokaryotic expression vector pET-30a (+) (Novagen) and sequenced to verify correctness, resulting in the expression plasmid used for transformation assay. The plasmid containing the target gene obtained above was transformed into cloning competent DH5α (Transgen, CD201-01) using the heat shock method (method described in the competent medium instruction manual), and the clone was selected to extract the plasmid for sequencing verification. The nucleotide sequence and amino acid sequence of the expressed protein are:

[0079] Table 1 Nucleotide and amino acid sequences of type III interferon proteins and stabilized type III interferon proteins

[0080]

[0081]

[0082]

[0083]

[0084]

[0085] Note: The bold character is the first amino acid G, the bold + underline character is the heparin binding sequence, and the underline character is the GS Linker.

[0086] 1.2 Protein expression

[0087] The correctly sequenced plasmid was transformed into expression competent BL21 (DE3) cells (Transgene, CD601-02) according to the competent instructions. Then, single colonies were randomly picked from the transformed plates and shaken. When the OD600 value was about 0.7, IPTG with a final concentration of 0.5 mM was added for induction. The cells were cultured at 37°C for about 4 h and the cells were collected for SDS-PAGE.

[0088] The results showed that when the heparin binding sequence was located at the N-terminus, the protein was basically not expressed, but when the heparin binding sequence was located at the C-terminus, all proteins were well expressed. Figure 1 shown.

[0089] 1.3 Isolation and purification of fusion protein

[0090] The cells were disrupted after expression and washed with disruption buffer (10mM Tris-HCl, 1mM EDTA, pH 6.5) at a V:m ratio of 10ml:1g. The suspension was homogenized using a homogenizer and the precipitate was collected by centrifugation. The precipitate was washed with wash buffer (10mM Tris-HCl, 1mM EDTA, 0.5% Tween 80, pH 6.5) at a V:m ratio of 10ml:1g. Stirring was continued until no particles were visible, and the precipitate was collected by centrifugation. The precipitate was lysed with lysis buffer (50mM Tris-HCl, 7M guanidine hydrochloride, 10mM DTT, pH 6.5) at a V:m ratio of 10ml:1g, and the protein supernatant was collected by centrifugation. The protein supernatant was renatured using renaturation buffer (100 mM Tris-Base, 0.5 M L-Arg HCl, 2 mM GSH, 0.5 mM GSSG, pH 8.0) to a concentration of approximately 0.1 mg / ml for 12-24 h at 16°C. The protein concentrate was then concentrated 10-fold using a 5 kDa ultrafiltration membrane.

[0091] The protein concentrate was diluted with 1 volume of pure water and used as the loading sample. The column was packed with Sepharose FF filler for SP chromatography, equilibrated with 20 mM phosphate (PB) pH 7.4, 0.05 M NaCl, and loaded. Different protein samples were eluted with eluents of different salt concentrations, as shown in Table 2. The entire elution peak fraction was collected by isocratic elution.

[0092] 16 proteins were obtained, corresponding to the protein molecules in Table 2 below.

[0093] Table 2 Proteins and corresponding eluents

[0094] molecular eluent M1 20 mM PB, pH 7.4, 0.2 M NaCl M1G 20mM PB, pH 7.4, 0.276M NaCl M1G-AR 20 mM PB, pH 7.4, 0.5 M NaCl M1G-G4S-AR 20 mM PB, pH 7.4, 0.5 M NaCl M1G-G4S-AR(N133L) 20 mM PB, pH 7.4, 0.5 M NaCl M1G-G4S-FGF18 20mM PB, pH 7.4, 0.276M NaCl M1G-G4S-AAMP 20mM PB, pH 7.4, 0.276M NaCl M1A 20mM PB, pH 7.4, 0.276M NaCl M1P 20mM PB, pH 7.4, 0.276M NaCl M1S 20mM PB, pH 7.4, 0.276M NaCl M4G 20mM PB, pH 7.4, 0.276M NaCl M8G 20mM PB, pH 7.4, 0.276M NaCl M9G 20mM PB, pH 7.4, 0.276M NaCl M4G-G4S-FGF18 20mM PB, pH 7.4, 0.276M NaCl M8G-G4S-FGF18 20mM PB, pH 7.4, 0.276M NaCl M9G-G4S-FGF18 20mM PB, pH 7.4, 0.276M NaCl

[0095] The purified proteins were analyzed for purity using an Agilent 1260 HPLC system using a C18 reverse-phase column. The purity of the purified proteins was approximately 95%. The molecular weights were determined by mass spectrometry and were consistent with the theoretical molecular weights.

[0096] Example 2 Peptide mapping method to detect the stability of each protein

[0097] The M1, M1G, and M1G-G4S-AR proteins prepared in Example 1 were each displaced into 20 mM NaH2PO4, pH 4.5 buffer, adjusted to a protein concentration of 0.5 mg / ml, and placed under illumination at 25°C for 7 days. The oxidation of the N-terminal M was examined by peptide mapping, as shown in Table 3. It was found that when the N-terminus was M (M1), the N-terminal oxidation ratio was 2.5% after 5 days of exposure to light at 25°C. However, when the N-terminus was G, A, P, or S, the N-terminal oxidation ratio was zero under the same conditions. This greatly improved the stability of the sample, making it possible to store the sample at room temperature.

[0098] Table 3 N-terminal stability of each protein

[0099] sample N-terminal oxidation / % M1 2.5 M1G 0 M1G-AR 0 M1G-G4S-AR 0 M1G-G4S-AR(N133L) 0 M1G-G4S-FGF18 0 M1G-G4S-AAMP 0 M1A 0 M1P 0 M1S 0 M4G 0 M8G 0 M9G 0 M4G-G4S-FGF18 0 M8G-G4S-FGF18 0 M9G-G4S-FGF18 0

[0100] Example 3 Heparin affinity experiments of various proteins

[0101] The affinity between the protein prepared above and heparin was detected by surface plasmon resonance (SPR) in Biacore 8K (GE).

[0102] First, each of the proteins prepared above was immobilized via amino groups onto a CM5 sensor chip (GE, BR100530). The analyte was heparin diluted in regeneration buffer at varying concentrations, increasing in 2-fold increments from a minimum of 0.195 nM. Zero concentration and reference channels were used for detection. Using SPR technology, the association and dissociation rates of the different proteins were measured and their binding affinities determined.

[0103] The results of the analysis using Biacore Insight Evaluation Software 2.0.15.12933 are shown in Table 4 below:

[0104] Table 4 SPR analysis results of various proteins and heparin

[0105]

[0106] Note: ka represents the association rate constant, kd represents the dissociation rate constant, KD represents the kd / ka value, Rmax represents the maximum response signal, chi represents the linear fit, and %Bound represents the capture protein response signal.

[0107] From the KD values in Table 4, it can be seen that the affinity level order is: M1G-G4S-AAMP>M1G-G4S-FGF18≈M1G-G4S-AR≈M1G-G4S-AR(N133L)>M1; Figure 2 It can be seen that the M1 response signal is extremely low, the linearity is poor, and the affinity to heparin is low.

[0108] Example 4 In vitro cell biological activity assay of each protein

[0109] HEK293-ISRE-Luc cells (purchased from China Food and Drug Inspection Institute) were seeded into 96-well plates at a density of 20,000 cells / well and cultured in a 37°C 5% CO2 incubator for 24 hours. The proteins prepared in Example 2 were diluted according to their respective concentrations, starting with 400 nmol / L, 6-fold dilutions at 7 points, and a 0-point control, for a total of 8 concentration points of protein samples. The culture medium was discarded, 50 μl of sample was added to each well, and after reacting with the cells in the incubator for 3 hours, the culture medium was discarded, and 300 μl / well culture medium was added to rinse the cells, the liquid was discarded, and the cells were repeatedly rinsed. Finally, 100 μl / well culture medium was added to the incubator and cultured for 23 hours. Cell lysate and luciferase substrate were added according to the instructions of the luciferase detection kit (Bright-GloTM Luciferase Assay System, Promega), and the chemiluminescence enzyme reader was used for measurement, and the EC values of each protein sample were recorded respectively. 50 Luciferase was detected (Promega, E2620), and the reading and four-parameter curve analysis were performed using Molecular Devices SpectraMax L. Figure 3 .

[0110] according to Figure 3 The EC values of each protein were calculated using the software SofeMax Pro v5.0.1. 50 For details, see Table 5 below:

[0111] Table 5 In vitro cell biological activity data of each protein

[0112]

[0113]

[0114] The above results indicate that the interferon activity is relatively increased after adding the heparin binding sequence.

[0115] Example 5 Pharmacokinetics in Mice

[0116] 90 Balb / c female mice (6-8 weeks old, Sibeifu (Beijing) Biotechnology Co., Ltd.) with a body weight of 18-22g were ordered. The mice were randomly divided into 3 groups: M1, M1G-G4S-AR and M1G-G4S-FGF18 groups, with 30 mice in each group. The drugs were administered by nebulization, with a concentration of 51μM and a nebulization volume of 10ml. After the nebulization, lung tissue samples were collected at various time points: 0 o'clock, 0.5h, 1h, 3h, 6h and 24h after administration. At each time point, 5 mice were randomly selected for lung tissue cross-sectioning. The lung tissue was weighed and PBS was added for quantitative homogenization. The drug concentration in the lung tissue was detected by ELISA. The results are shown in Table 6 and Table 6 below. Figure 4 As shown:

[0117] Table 6 Pharmacokinetic results of each protein in mice

[0118]

[0119] From the above results, it can be seen that there is no statistical difference in lung drug concentration between the M1G-G4S-AR and M1G-G4S-FGF18 groups; however, there are statistical differences between these two groups and the M1 group, indicating that the pharmacokinetic effect of interferon with the addition of heparin binding sequence in mice is significantly improved compared with interferon without the addition of heparin binding sequence.

[0120] Sequence Listing:

[0121]

[0122]

[0123]

[0124]

[0125] Sequence Listing <110> Hangzhou Xianweida Biotechnology Co., Ltd. <120> Stable type III interferon protein and its fusion protein <130> PF02223 <150> 2021103961265 <151> 2021-04-13 <160> 58 <170> PatentIn version 3.5 <210> 1 <211> 175 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 1 Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys Ser 1 5 10 15 Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala Leu 20 25 30 Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val Phe 35 40 45 Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro Val 50 55 60 Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala Ala 65 70 75 80 Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr Leu 85 90 95 His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro Thr 100 105 110 Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg Leu 115 120 125 Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser Val 130 135 140 Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val Ala 145 150 155 160 Asp Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 2 <211> 181 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 2 Gly Pro Val Pro Thr Ser Lys Pro Thr Thr Thr Gly Lys Gly Cys His 1 5 10 15 Ile Gly Arg Phe Lys Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys 20 25 30 Lys Ala Arg Asp Ala Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser 35 40 45 Cys Ser Ser Pro Val Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln 50 55 60 Val Arg Glu Arg Pro Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu 65 70 75 80 Lys Val Leu Glu Ala Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp 85 90 95 Gln Pro Leu His Thr Leu His His Ile Leu Ser Gln Leu Gln Ala Cys 100 105 110 Ile Gln Pro Gln Pro Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His 115 120 125 His Trp Leu His Arg Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly 130 135 140 Cys Leu Glu Ala Ser Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg 145 150 155 160 Asp Leu Lys Tyr Val Ala Asp Gly Asn Leu Cys Leu Arg Thr Ser Thr 165 170 175 His Pro Glu Ser Thr 180 <210> 3 <211> 200 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 3 Met Ala Ala Ala Trp Thr Val Val Leu Val Thr Leu Val Leu Gly Leu 1 5 10 15 Ala Val Ala Gly Pro Val Pro Thr Ser Lys Pro Thr Thr Thr Gly Lys 20 25 30 Gly Cys His Ile Gly Arg Phe Lys Ser Leu Ser Pro Gln Glu Leu Ala 35 40 45 Ser Phe Lys Lys Ala Arg Asp Ala Leu Glu Glu Ser Leu Lys Leu Lys 50 55 60 Asn Trp Ser Cys Ser Ser Pro Val Phe Pro Gly Asn Trp Asp Leu Arg 65 70 75 80 Leu Leu Gln Val Arg Glu Arg Pro Val Ala Leu Glu Ala Glu Leu Ala 85 90 95 Leu Thr Leu Lys Val Leu Glu Ala Ala Ala Gly Pro Ala Leu Glu Asp 100 105 110 Val Leu Asp Gln Pro Leu His Thr Leu His His Ile Leu Ser Gln Leu 115 120 125 Gln Ala Cys Ile Gln Pro Gln Pro Thr Ala Gly Pro Arg Pro Arg Gly 130 135 140 Arg Leu His His Trp Leu His Arg Leu Gln Glu Ala Pro Lys Lys Glu 145 150 155 160 Ser Ala Gly Cys Leu Glu Ala Ser Val Thr Phe Asn Leu Phe Arg Leu 165 170 175 Leu Thr Arg Asp Leu Lys Tyr Val Ala Asp Gly Asn Leu Cys Leu Arg 180 185 190 Thr Ser Thr His Pro Glu Ser Thr 195 200 <210> 4 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 4 Met Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 5 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 5 Met Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 6 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 6 Met Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Ser Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 7 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 7 Met Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Ser Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 8 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 8 Met Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 9 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 9 Met Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 10 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 10 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 11 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 11 Ala Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 12 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 12 Pro Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 13 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 13 Ser Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 14 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 14 Gly Gly Gly Gly Ser 1 5 <210> 15 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 15 Lys Arg Lys Lys Lys Gly Gly Lys Asn Gly Lys Asn Arg Arg Asn Arg 1 5 10 15 Lys Lys Lys Asn Pro 20 <210> 16 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 16 Lys Arg Lys Lys Lys Gly Gly Lys Leu Gly Lys Asn Arg Arg Asn Arg 1 5 10 15 Lys Lys Lys Asn Pro 20 <210> 17 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 17 Phe Thr Lys Lys Gly Arg Pro Arg Lys Gly Pro Lys Thr Arg Glu Asn 1 5 10 15 Gln Gln Asp Val His 20 <210> 18 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 18 Arg Arg Leu Arg Arg Met Glu Ser Glu Ser Glu Ser 1 5 10 <210> 19 <211> 531 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 19 atgaaaccga ccacgaccgg caaaggctgc catattggtc gctttaagtc gctgtcgccg 60 caggaactgg cgagcttcaa gaaagcccgt gatgccctgg aggaatcgct gaaactgaag 120 aactggagct gtagctcgcc ggtgttcccg ggcaactggg atctgcgtct gctgcaggtt 180 cgcgaacgtc cggttgcgct ggaagcggaa ctggcgctga ccctgaaagt gctggaagcg 240 gcagcgggtc cggcgctgga agatgttctg gatcagccgc tgcacaccct gcatcatatt 300 ctgtcgcagc tgcaggcgtg cattcaaccg cagccgaccg cgggcccgcg tccgcgcggc 360 cgtctgcatc actggctgca ccgtctgcag gaagccccga agaaagagtc ggcgggctgt 420 ctggaagcgt cggtgacctt caatctgttc cgtctgctga cccgtgatct gaaatacgtt 480 gcggaaggca atctgtctct gcgtacctcg acccacccgg aatcgaccta a 531 <210> 20 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 20 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac ctaa 534 <210> 21 <211> 597 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 21 atggggaagc gcaaaaaaaa aggcggcaaa aacggtaaaa atcgtcgtaa ccgtaagaaa 60 aaaaatccta aaccgaccac gaccggcaaa ggctgccata ttggtcgctt taagtcgctg 120 tcgccgcagg aactggcgag cttcaagaaa gcccgtgatg ccctggagga atcgctgaaa 180 ctgaagaact ggagctgtag ctcgccggtg ttcccgggca actgggatct gcgtctgctg 240 caggttcgcg aacgtccggt tgcgctggaa gcggaactgg cgctgaccct gaaagtgctg 300 gaagcggcag cgggtccggc gctggaagat gttctggatc agccgctgca caccctgcat 360 catattctgt cgcagctgca ggcgtgcatt caaccgcagc cgaccgcggg cccgcgtccg 420 cgcggccgtc tgcatcactg gctgcaccgt ctgcaggaag ccccgaagaa agagtcggcg 480 ggctgtctgg aagcgtcggt gaccttcaat ctgttccgtc tgctgacccg tgatctgaaa 540 tacgttgcgg aaggcaatct gtctctgcgt acctcgaccc acccggaatc gacctaa 597 <210> 22 <211> 597 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 22 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac caagcgcaaa 540 aaaaaaggcg gcaaaaacgg taaaaatcgt cgtaaccgta agaaaaaaaa tccttaa 597 <210> 23 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 23 atggggaagc gcaaaaaaaa aggcggcaaa aacggtaaaa atcgtcgtaa ccgtaagaaa 60 aaaaatcctg gtggtggtgg ttctaaaccg accacgaccg gcaaaggctg ccatattggt 120 cgctttaagt cgctgtcgcc gcaggaactg gcgagcttca agaaagcccg tgatgccctg 180 gaggaatcgc tgaaactgaa gaactggagc tgtagctcgc cggtgttccc gggcaactgg 240 gatctgcgtc tgctgcaggt tcgcgaacgt ccggttgcgc tggaagcgga actggcgctg 300 accctgaaag tgctggaagc ggcagcgggt ccggcgctgg aagatgttct ggatcagccg 360 ctgcacaccc tgcatcatat tctgtcgcag ctgcaggcgt gcattcaacc gcagccgacc 420 gcgggcccgc gtccgcgcgg ccgtctgcat cactggctgc accgtctgca ggaagccccg 480 aagaaagagt cggcgggctg tctggaagcg tcggtgacct tcaatctgtt ccgtctgctg 540 acccgtgatc tgaaatacgt tgcggaaggc aatctgtctc tgcgtacctc gacccacccg 600 gaatcgacct aa 612 <210> 24 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 24 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac cggtggtggt 540 ggttctaagc gcaaaaaaaa aggcggcaaa aacggtaaaa atcgtcgtaa ccgtaagaaa 600 aaaaatcctt aa 612 <210> 25 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 25 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac cggtggtggt 540 ggttctaagc gcaaaaaaaa aggcggcaaa ctgggtaaaa atcgtcgtaa ccgtaagaaa 600 aaaaatcctt aa 612 <210> 26 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 26 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac cggtggtggt 540 ggttctttca ccaaaaaagg ccgcccgcgc aaaggcccga aaacccgcga aaatcagcag 600 gatgttcatt aa 612 <210> 27 <211> 585 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 27 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac cggtggtggt 540 ggttctcgcc gcctgcgccg catggaaagc gaaagcgaaa gctaa 585 <210> 28 <211> 197 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 28 Gly Lys Arg Lys Lys Lys Gly Gly Lys Asn Gly Lys Asn Arg Arg Asn 1 5 10 15 Arg Lys Lys Lys Asn Pro Lys Pro Thr Thr Thr Gly Lys Gly Cys His 20 25 30 Ile Gly Arg Phe Lys Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys 35 40 45 Lys Ala Arg Asp Ala Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser 50 55 60 Cys Ser Ser Pro Val Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln 65 70 75 80 Val Arg Glu Arg Pro Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu 85 90 95 Lys Val Leu Glu Ala Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp 100 105 110 Gln Pro Leu His Thr Leu His His Ile Leu Ser Gln Leu Gln Ala Cys 115 120 125 Ile Gln Pro Gln Pro Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His 130 135 140 His Trp Leu His Arg Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly 145 150 155 160 Cys Leu Glu Ala Ser Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg 165 170 175 Asp Leu Lys Tyr Val Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr 180 185 190 His Pro Glu Ser Thr 195 <210> 29 <211> 197 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 29 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Lys Arg Lys Lys Lys Gly Gly Lys Asn Gly Lys Asn Arg Arg Asn Arg 180 185 190 Lys Lys Lys Asn Pro 195 <210> 30 <211> 202 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 30 Gly Lys Arg Lys Lys Lys Gly Gly Lys Asn Gly Lys Asn Arg Arg Asn 1 5 10 15 Arg Lys Lys Lys Asn Pro Gly Gly Gly Gly Ser Lys Pro Thr Thr Thr 20 25 30 Gly Lys Gly Cys His Ile Gly Arg Phe Lys Ser Leu Ser Pro Gln Glu 35 40 45 Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala Leu Glu Glu Ser Leu Lys 50 55 60 Leu Lys Asn Trp Ser Cys Ser Ser Pro Val Phe Pro Gly Asn Trp Asp 65 70 75 80 Leu Arg Leu Leu Gln Val Arg Glu Arg Pro Val Ala Leu Glu Ala Glu 85 90 95 Leu Ala Leu Thr Leu Lys Val Leu Glu Ala Ala Ala Gly Pro Ala Leu 100 105 110 Glu Asp Val Leu Asp Gln Pro Leu His Thr Leu His His Ile Leu Ser 115 120 125 Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro Thr Ala Gly Pro Arg Pro 130 135 140 Arg Gly Arg Leu His His Trp Leu His Arg Leu Gln Glu Ala Pro Lys 145 150 155 160 Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser Val Thr Phe Asn Leu Phe 165 170 175 Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val Ala Glu Gly Asn Leu Ser 180 185 190 Leu Arg Thr Ser Thr His Pro Glu Ser Thr 195 200 <210> 31 <211> 202 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 31 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Gly Gly Gly Gly Ser Lys Arg Lys Lys Lys Gly Gly Lys Asn Gly Lys 180 185 190 Asn Arg Arg Asn Arg Lys Lys Lys Asn Pro 195 200 <210> 32 <211> 202 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 32 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Gly Gly Gly Gly Ser Lys Arg Lys Lys Lys Gly Gly Lys Leu Gly Lys 180 185 190 Asn Arg Arg Asn Arg Lys Lys Lys Asn Pro 195 200 <210> 33 <211> 202 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 33 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Gly Gly Gly Gly Ser Phe Thr Lys Lys Gly Arg Pro Arg Lys Gly Pro 180 185 190 Lys Thr Arg Glu Asn Gln Gln Asp Val His 195 200 <210> 34 <211> 193 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 34 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Gly Gly Gly Gly Ser Arg Arg Leu Arg Arg Met Glu Ser Glu Ser Glu 180 185 190 Serum <210> 35 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 35 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 36 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 36 Ala Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 37 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 37 Pro Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 38 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 38 Ser Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 39 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 39 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 40 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 40 Ala Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 41 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 41 Pro Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 42 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 42 Ser Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 43 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 43 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 44 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 44 Ala Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 45 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 45 Pro Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 46 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 46 Ser Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 <210> 47 <211> 202 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 47 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Glu Gly Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Gly Gly Gly Gly Ser Phe Thr Lys Lys Gly Arg Pro Arg Lys Gly Pro 180 185 190 Lys Thr Arg Glu Asn Gln Gln Asp Val His 195 200 <210> 48 <211> 202 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 48 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Cys Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Gly Gly Gly Gly Ser Phe Thr Lys Lys Gly Arg Pro Arg Lys Gly Pro 180 185 190 Lys Thr Arg Glu Asn Gln Gln Asp Val His 195 200 <210> 49 <211> 202 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 49 Gly Lys Pro Thr Thr Thr Gly Lys Gly Cys His Ile Gly Arg Phe Lys 1 5 10 15 Ser Leu Ser Pro Gln Glu Leu Ala Ser Phe Lys Lys Ala Arg Asp Ala 20 25 30 Leu Glu Glu Ser Leu Lys Leu Lys Asn Trp Ser Cys Ser Ser Pro Val 35 40 45 Phe Pro Gly Asn Trp Asp Leu Arg Leu Leu Gln Val Arg Glu Arg Pro 50 55 60 Val Ala Leu Glu Ala Glu Leu Ala Leu Thr Leu Lys Val Leu Glu Ala 65 70 75 80 Ala Ala Gly Pro Ala Leu Glu Asp Val Leu Asp Gln Pro Leu His Thr 85 90 95 Leu His His Ile Leu Ser Gln Leu Gln Ala Cys Ile Gln Pro Gln Pro 100 105 110 Thr Ala Gly Pro Arg Pro Arg Gly Arg Leu His His Trp Leu His Arg 115 120 125 Leu Gln Glu Ala Pro Lys Lys Glu Ser Ala Gly Cys Leu Glu Ala Ser 130 135 140 Val Thr Phe Asn Leu Phe Arg Leu Leu Thr Arg Asp Leu Lys Tyr Val 145 150 155 160 Ala Asp Ala Asn Leu Ser Leu Arg Thr Ser Thr His Pro Glu Ser Thr 165 170 175 Gly Gly Gly Gly Ser Phe Thr Lys Lys Gly Arg Pro Arg Lys Gly Pro 180 185 190 Lys Thr Arg Glu Asn Gln Gln Asp Val His 195 200 <210> 50 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 50 atggcgaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac ctaa 534 <210> 51 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 51 atgccgaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac ctaa 534 <210> 52 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 52 atgagcaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtc tctgcgtacc tcgacccacc cggaatcgac ctaa 534 <210> 53 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 53 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtg tctgcgtacc tcgacccacc cggaatcgac ctaa 534 <210> 54 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 54 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag cgaatctgtg tctgcgtacc tcgacccacc cggaatcgac ctaa 534 <210> 55 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 55 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag cgaatctgtc tctgcgtacc tcgacccacc cggaatcgac ctaa 534 <210> 56 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 56 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag gcaatctgtg tctgcgtacc tcgacccacc cggaatcgac cggtggtggt 540 ggttctttca ccaaaaaagg ccgcccgcgc aaaggcccga aaacccgcga aaatcagcag 600 gatgttcatt aa 612 <210> 57 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 57 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag cgaatctgtg tctgcgtacc tcgacccacc cggaatcgac cggtggtggt 540 ggttctttca ccaaaaaagg ccgcccgcgc aaaggcccga aaacccgcga aaatcagcag 600 gatgttcatt aa 612 <210> 58 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: artificially synthesized sequence <400> 58 atggggaaac cgaccacgac cggcaaaggc tgccatattg gtcgctttaa gtcgctgtcg 60 ccgcaggaac tggcgagctt caagaaagcc cgtgatgccc tggaggaatc gctgaaactg 120 aagaactgga gctgtagctc gccggtgttc ccgggcaact gggatctgcg tctgctgcag 180 gttcgcgaac gtccggttgc gctggaagcg gaactggcgc tgaccctgaa agtgctggaa 240 gcggcagcgg gtccggcgct ggaagatgtt ctggatcagc cgctgcacac cctgcatcat 300 attctgtcgc agctgcaggc gtgcattcaa ccgcagccga ccgcgggccc gcgtccgcgc 360 ggccgtctgc atcactggct gcaccgtctg caggaagccc cgaagaaaga gtcggcgggc 420 tgtctggaag cgtcggtgac cttcaatctg ttccgtctgc tgacccgtga tctgaaatac 480 gttgcggaag cgaatctgtc tctgcgtacc tcgacccacc cggaatcgac cggtggtggt 540 ggttctttca ccaaaaaagg ccgcccgcgc aaaggcccga aaacccgcga aaatcagcag 600 gatgttcatt aa 612

Claims

1. A stable type III interferon protein having the amino acid sequence of SEQ ID NO.

10.

2. A stable type III interferon protein fusion protein having the structure A-Ln-B, wherein: A is the amino acid sequence of SEQ ID NO. 10, L is a linker, the linker is a glycine-rich peptide, the linker is a peptide having GGGGS (SEQ ID NO. 14), n is 0 to 6, B is a binding sequence targeting epithelial cells, wherein the binding sequence targeting epithelial cells is a heparin binding sequence, and the heparin binding sequence is any one selected from the following: KRKKKKGGKNGKNRRRNRKKKNP (SEQ ID NO. 15); KRKKKKGGKLGKNRRRNRKKKNP (SEQ ID NO. 16); FTKKGRPRKGPKTRENQQDVH (SEQ ID NO. 17); RRLRRMESESES (SEQ ID NO. 18).

3. The fusion protein according to claim 2, wherein The epithelial cell targeting binding sequence is located at the C-terminus of the stabilized type III interferon protein.

4. The fusion protein according to claim 3, wherein The linker is located at the C-terminus of the stable type III interferon protein.

5. The fusion protein according to claim 2, wherein n is 1.

6. The fusion protein according to claim 2, wherein The heparin binding sequence is the sequence shown in SEQ ID NO. 15 or SEQ ID NO.

17.

7. The fusion protein according to claim 2, wherein The amino acid sequence is selected from: SEQ ID NO. 29, 31~34.

8. The fusion protein according to claim 2, wherein the fusion protein is fused to other polypeptides or proteins at the N-terminus or the C-terminus.

9. A conjugate of a stabilized type III interferon protein according to claim 1 and a fusion protein according to any one of claims 2 to 8, wherein the type III interferon protein and the fusion protein thereof are conjugated to polyethylene glycol.

10. The conjugate according to claim 9, wherein the polyethylene glycol is a linear or branched polyethylene glycol.

11. The conjugate according to claim 10, wherein the linear or branched polyethylene glycol is mPEG propionaldehyde of 20 kD, 30 kD or 40 kD.

12. A polynucleotide encoding the stabilized type III interferon protein of claim 1 or the fusion protein of any one of claims 2 to 8.

13. A vector comprising the polynucleotide of claim 12.

14. A host cell comprising the polynucleotide of claim 12 or the vector of claim 13.

15. A method for preparing the stabilized type III interferon protein of claim 1 or the fusion protein of any one of claims 2 to 8, comprising expressing the stabilized type III interferon protein or the fusion protein in the host cell of claim 14, and isolating the stabilized type III interferon protein or the fusion protein from a culture of the host cell. 16 . A pharmaceutical composition comprising the stabilized type III interferon protein of claim 1 , the fusion protein of any one of claims 2 to 8 , or the conjugate of claims 9 to 11 , and a pharmaceutically acceptable carrier. 17 . A kit comprising the stabilized type III interferon protein of claim 1 , the fusion protein of any one of claims 2 to 8 , the conjugate of claims 9 to 11 , or the pharmaceutical composition of claim 16 .

18. A product comprising the stabilized type III interferon protein of claim 1, the fusion protein of any one of claims 2 to 8, the conjugate of claims 9 to 11, or the pharmaceutical composition of claim 16.

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