Vaccine composition against severe fever with thrombocytopenia syndrome virus

Abstract

The present invention provides a vaccine composition against SFTS virus using Gn or Gc protein or NP protein or the mixture thereof, based on the experimental results that the recombinant protein of the extracellular domain of Gn or Gc protein of STFS fused to the Fc site of human antibody, NP protein (full-length sequence protein) or the mixture thereof provides protective immunity in type I interferon receptor gene knockout (KO) mice.

Description

Technical Field

[0001] The present invention relates to a vaccine composition for severe fever with thrombocytopenia syndrome virus. Background Art

[0002] Severe fever with thrombocytopenia syndrome (SFTS) is an infectious disease caused by infection with the fever with thrombocytopenia syndrome virus (SFTS virus, SFTSV). The genome of STFSV includes three segmented ribonucleic acids (RNA), which encode RNA polymerase (RdRp), two envelope glycoproteins (Gn, Gc), nucleocapsid (NP) and non-structural protein (NS). When infected with the virus, it is accompanied by fever, gastrointestinal complications, leukopenia and thrombocytopenia symptoms, and its mortality rate reaches 5-20% (Lancet Infect Dis 2018, 18, 1127-1137, doi: 10.1016 / S1473-3099 (18) 30293-7). So far, there is no therapeutic agent, and vaccine development is underway.

[0003] The development of STFS vaccines is still in the early stages of research. Recently, reports have pointed out that vaccines using recombinant viral vectors or deoxyribonucleic acid (DNA) vaccines can provide protective immunity in infected animal models (NPJ Vaccines 2019, 4, 5, doi: 10.1038 / s41541-018-0096-y; Nat Commun 2019, 10, 3836, doi: 10.1038 / s41467-019-11815-4). It has been confirmed that vaccines using NS can induce the formation of antibodies, but they have not been able to provide protective immunity in mouse infection models (Viral Immunol 2015, 28, 113-122, doi: 10.1089 / vim.2014.0100). It has been reported that micromanipulation of DNA encoding NP and NS genes can induce specific T cell responses, but it has not yet been determined whether protective immunity can be provided in animal infection models (Colloids Surf B Biointerfaces 2017, 159, 54-61, doi: 10.1016 / j.colsurfb.2017.07.059). Recently, a paper reported that a single immunization with a recombinant virus expressing Gn and Gc proteins (live attenuated recombinant vesicular stomatitis virus, rVSV) can induce the formation of neutralizing antibodies, and can induce perfect protective immunity in type I interferon receptor knockout (KO) mice infected with a lethal dose of STFSV (NPJ Vaccines 2019, 4, 5, doi: 10.1038 / s41541-018-0096-y). In addition, it has been reported that when DNA expressing five genes of SFTSV is immunized simultaneously, two DNAs encoding Gn and Gc are immunized simultaneously, and three DNAs encoding NP, NS, and RdRp are immunized simultaneously, it can provide immune protection against lethal infection of elderly weasels (over four years old) susceptible to SFTSV (Nat Commun 2019, 10, 3836, doi: 10.1038 / s41467-019-11815-4). However, there are no reports so far indicating the degree of protective immunity that can be provided when immunizing each structural protein (Gn, Gc, NP) separately, and no research results related to the efficacy of subunit protein vaccines using the above proteins have been published.

[0004] The present invention discloses the vaccine efficacy of Gn protein, Gc protein and NP protein. DETAILED DESCRIPTION OF THE INVENTION

[0006] Technical issues

[0007] The object of the present invention is to provide a vaccine composition against SFTS virus using Gn protein, Gc protein and NP protein.

[0008] Other objects or specific objects of the present invention will be described in the following contents.

[0009] Means of solving the problem

[0010] As shown in the following examples, the present inventors compared and verified whether the recombinant units (Gn-Fc and Gc-Fc) in which the extracellular domain of Gn or Gc protein is fused to the Fc site of human antibody and NP protein (full-length sequence protein) can be used alone to immunize type I interferon receptor KO mice and provide immune protection. As a result, all mice immunized with Gc-Fc protein died, but their survival time showed a statistically significant increase compared with the control group (group immunized with Fc only), and 50% of mice immunized with Gn-Fc protein survived, and 60% of mice immunized with NP protein survived, thereby confirming that each has a significant protective immunity providing effect. In addition, when type I interferon receptor KO mice were immunized with a mixture of Gn-Fc and Gc-Fc and NP to confirm whether their protective immunity was improved, 80% of the mice survived when immunized with a mixture of Gc-Fc and NP, 70% of the mice survived when immunized with a mixture of Gn-Fc and NP, and 60% of the mice survived when immunized with a mixture of Gn-Fc and Gc-Fc and NP. This confirmed that, overall, the survival rate was improved in the case of mixed immunization compared to single immunization.

[0011] The present invention is provided based on the experimental results as described above. The vaccine composition of the present invention relates to a vaccine composition against SFTS virus which comprises the extracellular domain of Gc protein, the extracellular domain of Gn protein, the full-length NP protein or a mixture of two or more thereof as an active ingredient.

[0012] In the present invention, in particular, the extracellular domain of the Gc protein or the extracellular domain of the Gn protein can be fused to the Fc site of a human antibody in order to increase the half-life in vivo, and in the case of fusion with Fc, in order to prevent the interference of each fusion protein with the formation of a three-dimensional structure between each other, a hinge sequence of 2 to 15 amino acids can be configured. In addition, in order to produce the above-mentioned protein using a host cell by genetic recombination, as described in the following embodiments, separation and purification can be more easily achieved by configuring a signal peptide sequence at the N-terminus. Because the above-mentioned sequence is removed when the corresponding protein is secreted outside the cell, it does not exist in the separated and purified protein.

[0013] In this specification, "vaccine" refers to a vaccine that prevents infection or reinfection with a corresponding pathogen, alleviates or eliminates symptoms caused by a corresponding pathogen, or substantially or completely eliminates a corresponding pathogen or a disease caused by a corresponding pathogen by inducing an immune response to the corresponding pathogen in a host, i.e., a human. Therefore, the vaccine composition of the present invention can be administered to humans for the purpose of prevention before infection with the corresponding pathogen, or for the purpose of treatment after infection with the corresponding pathogen. The "immune response" refers to a humoral immune response, a cellular immune response, or both.

[0014] In addition, in this specification, "active ingredient" refers to a component that can induce an immune response alone or can induce an immune response together with a carrier that cannot induce an immune response by itself. Therefore, the active ingredient itself does not necessarily need to have immunogenicity (the ability to induce an immune response). In the present invention, it is preferable that the active ingredient is purified, but it is not necessary. Among them, "purified" refers to the state in which the cell components or its culture fluid components (which belong to contaminants) in the original organism where the corresponding target substance exists (i.e., the microorganism that undergoes morphological transformation in the present invention) are substantially reduced or removed. The state in which the contaminants are substantially reduced or removed refers to a purity of more than 50%, preferably more than 90%, and more preferably more than 99%. Purity can be evaluated by methods known to related industries such as chromatographic columns, gel electrophoresis, and immunological analysis, and the purification method can also use methods known to related industries described later.

[0015] The vaccine composition of the present invention may be prepared in any suitable and pharmaceutically acceptable formulation, for example, in the form of a ready-to-administer solution or suspension, or in the form of a concentrated stock solution suitable for dilution prior to administration, or in a reconstituted form such as a freeze-dried, freeze-dried or frozen preparation.

[0016] The vaccine composition of the present invention may include a pharmaceutically acceptable carrier when formulated.

[0017] The carrier as described above may generally include diluents, excipients, stabilizers and preservatives, etc. As appropriate diluents, non-aqueous solvents such as propylene glycol, polyethylene glycol and vegetable oils (such as olive oil and peanut oil) or aqueous solvents such as saline (preferably 0.8% saline) and water containing a buffer medium (preferably 0.05M phosphate buffer) may be used as examples, as appropriate excipients, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, anhydrous skim milk, glycerol, propylene, ethylene glycol, water and ethanol may be used as examples, and as appropriate stabilizers, carbohydrates such as sorbitol, mannitol, starch, hydroxyethyl alcohol, dextran, glutamic acid and glucose or proteins such as animal, plant or microbial proteins (such as milk powder, serum albumin and casein) may be used as examples. As suitable preservatives, thimerosal, thimerosal, gentamicin, neomycin, nystatin, amphotericin B, tetracycline, penicillin, streptomycin and polymyxin B can be exemplified.

[0018] The vaccine composition of the present invention may also contain an antigen adjuvant. The antigen adjuvant may be composed of one or more substances that can strengthen the immune response to the antigen. The adjuvant may function as a tissue reservoir for slowly releasing the antigen and / or as a lymphatic system activator for non-specifically strengthening the immune response (Hood et al., Immunology, Second Ed., 1984, benjamin / Cummings: Menlo Park, California, p. 384). The antigen adjuvant may be, for example, Freund's complete adjuvant, Freund's incomplete adjuvant, saponin, gel-like aluminum adjuvant, surfactant (such as lysolecithin, polyether glycol, polyanion, peptide, oil, and hydrocarbon emulsion), vegetable oil (such as cottonseed oil, peanut oil or corn oil), vitamin E acetate, etc.

[0019] Aluminum adjuvants are currently the most widely used antigen adjuvants that can be used in humans. The aluminum adjuvants mentioned above are used in the form of gels of aluminum salts such as aluminum phosphate, potassium aluminum sulfate, and aluminum hydroxide. It is known that the aluminum adjuvants mentioned above can enhance immune responses by inducing Th2-type immune responses (Sokolovska A et al., Vaccine. 2007 Jun 625 (23): 4575-85; O'Hagan DT AND Rappuoli R., Pharm Res. 2004 Sep; 21 (9): 1519-30). Invivogen's Alhydrogel used in the embodiments of the present invention is a colloidal suspension of aluminum hydroxide. The aluminum adjuvant as described above can be directly manufactured and used according to the manufacturing method known in the relevant industry (R. Bomford. Immunological Adjuvants and Vaccines. NATO ASI Series 1989; 179: 35-41; Vogel FR AND Powell MF. Pharm Biotechnol. 1995; 6: 141-228.; Derek T. O'Hagan, Methods in Molecular Medicine. 2000; April 15; 42: 65-90), or purchased from the market. Examples of the products available on the market include the Alhydrogel 2% product of Invivogen used in the following examples, such as the Aluminum hydroxide Gel product of Sigma and the AlhydrogelTM product of BRENNTAG.

[0020] The vaccine composition of the present invention can be manufactured in any unit volume. The unit volume refers to the amount of active ingredients and pharmaceutically acceptable carriers contained in a single product packaged for the purpose of administering to humans more than once. The appropriate amount of the active ingredient and carrier as described above is the amount that can function as a vaccine when the vaccine composition of the present invention is used for more than one vaccination, and the amount as described above can be determined by non-clinical or clinical methods within the general ability of relevant practitioners.

[0021] The vaccine composition of the present invention can preferably be administered by non-oral routes such as rectal, percutaneous, intravenous, intraarterial, intramuscular, intradermal, subcutaneous, intraperitoneal, and intraventricular injections.

[0022] The vaccine composition of the present invention can be administered via a regulated release system. For example, as the regulated release system described above, liposomes, implantable osmotic pumps, and transdermal patches can be used as examples. Preferably, the active ingredient can be delivered by liposomes. For the content of delivering the active ingredient by liposomes, please refer to the literature [Langer, Science 249: 1527-1533 (1990)] and the literature [Treat at al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (des.), Liss, New York, pp. 353-365 (1989)], etc. For other delivery methods of active ingredients, please refer to the literature [Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987)], literature [Buchwald et al., Surgery 88:507 (1980)], literature [Saudek et al., N. Engl. J. Med. 321:574 (1989)], literature [Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974)], literature [Controlled Drug Bioavailability. Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984)], literature [Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23: 61 (1983)], reference [During et al., Ann. Neurol. 25: 351 (1989)], and reference [Howard et al., J. Neurosurg. 71: 105 (1989)], etc. The above-mentioned references are regarded as part of the present description.

[0023] The dosage of the vaccine composition of the present invention can be determined by medical personnel based on the patient's characteristics such as age, weight, gender, symptoms, complications, and whether or not the patient suffers from other diseases.

[0024] Furthermore, the time interval and frequency of administration can be determined according to the dosage form used or the half-life of the active ingredient in the blood.

[0025] As described above, the dosage, the time interval between dosages, and the number of dosages should depend on the judgment of the medical staff and be determined by the individual, but generally, an appropriate dosage should be determined within the range of about 0.1 to 10 mg per kg of individual body weight per day, the dosage interval should be determined within 3 to 10 days, and the number of dosages should be determined within the range of 1 to 5 times.

[0026] Each protein used as an active ingredient in the vaccine composition of the present invention can be produced by using the gene encoding it through DNA recombinant technology known in the relevant industry. The method as described above includes: (i) producing an expression vector containing a gene for expressing the gene encoding the protein; (ii) transforming the expression vector in a host cell; (iii) culturing the transformed host cell; and (iv) separating and purifying the protein from the culture. In this specification, the gene encoding the protein is disclosed, and the method of producing the corresponding protein by using the gene through DNA recombinant technology, specifically, the expression vector composition or recombinant DNA technology containing a signal peptide sequence of a regulatory sequence such as a promoter or terminator, a selection marker, etc., the transformation method, the composition of the culture medium, and the culture method after the transformation, etc., have accumulated a considerable amount of literature in the relevant industry, and these documents can be referred to for various related matters. Specifically, reference may be made to the literature (Nucl. Acid Res. 14: 5399-5467, 1986), literature (Tet. Lett. 27: 5575-5578, 1986), literature (Nucl. Acid Res. 4: 2557, 1977), literature (Lett., 28: 2449, 1978), literature (Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press (2001)), literature (FM Ausubel et al, Current Protocols in Molecular Biology, John Wiley & Sons, Inc. (1994)), literature (Marston, F (1987) DNA Cloning Techniques), literature (Cohen, SN et al., Proc. Natl. Acad. Sci. USA, 9: 2110-2114 (1973)), literature (Hanahan, D., J. Mol. Biol., 166: 557-580 (1983)), literature (Dower, WJ et al., Nucleic. Acids Res., 16: 6127-6145 (1988))(Capecchi, MR, Cell, 22: 479 (1980)), literature (Graham, FL et al., Virology, 52: 456 (1973)), literature (Neumann, E. et al., EMBO J., 1: 841 (1982)), literature (Wong, TKet al., Gene, 10: 87 (1980)), literature (Gopal, Mol. Cell Biol., 5: 1188-1190 (1985)), literature (Yang et al., Proc. Natl. Acad. Sci., 87: 9568-9572 (1990)) (Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory (1982)), literature (Hitzeman et al., J. Biol. Chem., 255, 12073-12080 (1980)), literature (Luchansky et al Mol. Microbiol. 2, 637-646 (1988)), literature (Sambrook, Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory Press, US (1989)), literature (Ausubel et al., Current Protocols in Molecular Biology, Jon Willey & Sons, US (1993)), literature (Sambrook, J. & Russel, D., Molecular Cloning, A LABORATORY MANUAL, Cold Spring Harbor Laboratory Press, Vol. 1 and Vol. 2 published on January 15, 2001), etc.

[0027] Effects of the Invention

[0028] As described above, the present invention can provide a vaccine composition against SFTS virus comprising, as an active ingredient, the extracellular domain of Gc protein, the extracellular domain of Gn protein, the full-length NP protein, or a mixture thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a schematic diagram illustrating the structure and molecular weight of the Gc protein, Gn protein or NP protein, which is the active ingredient of the vaccine composition of the present invention.

[0030] Figure 2 It is the result of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the active ingredient of the vaccine composition of the present invention, namely, Gc protein, Gn protein or NP protein.

[0031] Figure 3It is based on the induction of specific antibodies immunized with Gn and Gc antigens and the results of virus neutralization ability test using immune serum (a: antibody titer against Gn and Gn protein, b and c: neutralization ability analysis test against SFTS virus, FRNT: lesion reduction neutralization titer).

[0032] Figure 4 The results are as follows: type I interferon receptor KO mice were immunized with Gc protein, Gn protein or NP protein and their survival rate and body weight changes were measured.

[0033] Figure 5 The results are as follows: type I interferon receptor KO mice were immunized with a mixture of Gc protein, Gn protein and / or NP protein and their survival rate and body weight changes were measured. DETAILED DESCRIPTION

[0034] Next, the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited by the Examples.

[0035] <Example 1> Production of viral proteins for use as vaccine antigens

[0036] <Example 1-1> Amino Acid Sequences of Viral Proteins Used as Vaccine Antigens

[0037] The amino acid information on GenBank was used to produce Gn protein, Gc protein and NP protein used as vaccine antigens. The amino acid information on GenBank is GenBank ID: ATW63054.1 (Gn protein), GenBank ID: AOO85591.1 (Gc protein) and GenBank ID: AJO16084.1 (NP protein).

[0038] Gn and Gc proteins only use the extracellular domain (sequence numbers 1 and 2, respectively) excluding the signal peptide, transmembrane domain, and cytoplasmic domain, while NP protein uses the full-length sequence (sequence number 3).

[0039] <Example 1-2> Cloning and expression of viral protein genes

[0040] After synthesizing the genes encoding the Gn and Gc proteins (extracellular domain) (sequence numbers 4 and 5, respectively), Figure 1In the manner shown, the signal peptide gene (sequence number 6) of the Ig kappa light chain was fused to the N-terminal and the recombinant gene of the Fc domain (domain) gene (sequence number 7) of the human IgG1 CH2-CH3 site was fused to the C-terminal via the hinge region (sequence number 8) and cloned into the pCEP4 mammalian expression vector. The gene (sequence number 9) encoding the NP protein with 6His (for purification) connected to the N-terminus and the C-terminus was cloned into the pET28a vector. The cloned genes were introduced into HEK93T cells (Gn, Gc) and E.coli BL21 (NP) respectively to induce protein expression, and purified using a protein A / G column and a nickel-nitrilotriacetic acid column, respectively. The size and purity of the purified protein were confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Figure 2 ).

[0041] <Example 2> Immunization and Antibody Analysis of SFTS Virus Protein

[0042] Type I interferon receptor KO mice (n=4) were immunized three times at intervals of two weeks using Gn-Fc (SEQ ID NO. 10, the DVAQAA amino acid sequence at the N-terminus of SEQ ID NO. 10 was inserted for cloning), Gc-Fc (SEQ ID NO. 11, the DVAQAA amino acid sequence at the N-terminus of SEQ ID NO. 10 was inserted for cloning), NP (SEQ ID NO. 12), and human Fc (20ug / mouse+Alum, Gn-Fc, Gc-Fc, Fc; 10ug / mouse+Alum, NP). Two weeks after the third immunization, blood was collected and the antibody titers of Gn, Gc, and NP were measured by enzyme-linked immunosorbent assay (ELISA) ( Figure 3a). It was confirmed that high levels of antibodies were generated against each antigen (average titer of anti-Gn or anti-Gc IgG = 1:4,096, average titer of anti-NP IgG = 1:24,576; n = 4 / group), and in particular, high antibody titers were observed to be induced against the NP antigen. In order to confirm the virus neutralizing ability of antibodies generated against viral glycoproteins (Gn and Gc), the STFS virus was reacted with the immune serum and infected into Vero cells, and then a focus reduction assay was performed ( Figure 3 b and c). It was confirmed that although almost similar levels of antibodies were generated, the serum obtained by immunization with Gn-Fc formed a relatively higher neutralizing antibody titer than the serum obtained by immunization with Gc-Fc, and had a significantly higher level of virus neutralization ability than the serum obtained by immunization with only Fc in the two immune sera (neutralizing antibody titer: Gn-Fc: 929±1134, Gc-Fc: 209±140, Fc: 42±29; n=4 / group, mean titer±SD).

[0043] <Example 3> Protective immune effect of SFTS virus protein immunity

[0044] Type I interferon receptor KO mice (n=4-6) were immunized three times with Gn-Fc, Gc-Fc, NP, and Fc at intervals of two weeks (20ug / mouse+Alum, Gn-Fc, Gc-Fc, Fc; 10ug / mouse+Alum, NP). 3 The mice were infected subcutaneously with 100 foci-forming units (FFU) of SFTSV. In addition, the daily weight changes and mortality were observed ( Figure 4 ).

[0045] The weight of mice immunized with phosphate buffered saline (BPS) (control group) gradually decreased and all died until the sixth day of infection. The weight of mice immunized with Fc (control group) gradually decreased from the second day of infection and all died until the fifth day of infection. The weight of mice immunized with Gc-Fc gradually decreased from the fourth day of infection and all died until the eighth day of infection. The weight of mice immunized with Gn-Fc gradually decreased from the third day of infection and half of them died until the eighth day of infection, but the remaining mice gradually recovered their weight and survived. The weight of mice immunized with NP gradually decreased from the third day of infection and 40% died until the eighth day, but the remaining 60% of the mice gradually recovered their weight and survived. Statistical analysis (Log-rank (Mantel-Cox) Test) confirmed that the vaccine-based survival extension effect (Gc-Fc group: p = 0.0046) or mortality reduction effect (Gn-Fc group: p = 0.0054, NP group: p = 0.0023) showed a statistically significant improvement compared with the control group.

[0046] <Example 4> Protective immune effect of mixed immunization with SFTS virus proteins

[0047] Type I interferon receptor KO mice (n=7) were immunized three times at intervals of two weeks using a mixture of Gn-Fc and NP (Gn+NP, mixing ratio: 10ug+10ug), a mixture of Gc-Fc and NP (Gc+NP, mixing ratio: 10ug+10ug), a mixture of Gn-Fc and Gc-Fc and NP (Gn+Gc+NP, mixing ratio: 10ug+10ug), and phosphate buffered saline (BPS) (control) (20 or 30ug / mouse+Alum). Two weeks after the third immunization, 10 3 The mice were infected subcutaneously with 100 foci-forming units (FFU) of SFTSV. In addition, the daily weight changes and mortality were observed ( Figure 5 ).

[0048] The weight of mice immunized with phosphate buffered saline (BPS) (control group) gradually decreased and all died until the sixth day of infection. The weight of mice immunized with Gn+NP gradually decreased from the fourth day of infection, 14.3% died on the fifth day of infection, and 28.6% died until the sixth day of infection, but the remaining mice gradually recovered their weight and survived. The weight of mice immunized with Gc+NP gradually decreased from the fifth day of infection and 14.3% died on the sixth day of infection, but the remaining mice gradually recovered their weight and survived. The weight of mice immunized with Gn+Gc+NP gradually decreased from the fourth day of infection and 42.9% died on the seventh day of infection, but the remaining mice gradually recovered their weight and survived.

[0049] Statistical analysis (Log-rank (Mantel-Cox) Test) confirmed that the vaccine-based survival extension effect (Gc-Fc group: p = 0.0046) or mortality reduction effect (Gn-Fc group: p = 0.0054, NP group: p = 0.0023) showed a statistically significant improvement compared with the control group. SEQUENCE LISTING <110> Seoul National University Industry Cooperation Group <120> Vaccine composition against severe fever with thrombocytopenia syndrome virus <130> P22111831WP <160> 12 <170> PatentIn version 3.5 <210> 1 <211> 438 <212> PRT <213> Unknown <220> <223> SFTS virus Gn protein extracellular domain <400> 1 Asp Ser Gly Pro Ile Ile Cys Ala Gly Pro Ile His Ser Asn Lys Ser 1 5 10 15 Ala Asp Ile Pro His Leu Leu Gly Tyr Ser Glu Lys Ile Cys Gln Ile 20 25 30 Asp Arg Leu Ile His Val Ser Ser Trp Leu Arg Asn His Ser Gln Phe 35 40 45 Gln Gly Tyr Val Gly Gln Arg Gly Gly Arg Ser Gln Val Ser Tyr Tyr 50 55 60 Pro Ala Glu Asn Ser Tyr Ser Arg Trp Ser Gly Leu Leu Ser Pro Cys 65 70 75 80 Asp Ala Asp Trp Leu Gly Met Leu Val Val Lys Lys Ala Lys Gly Ser 85 90 95 Asp Met Ile Val Pro Gly Pro Ser Tyr Lys Gly Lys Val Phe Phe Glu 100 105 110 Arg Pro Thr Phe Asp Gly Tyr Val Gly Trp Gly Cys Gly Ser Gly Lys 115 120 125 Ser Arg Thr Glu Ser Gly Glu Leu Cys Ser Ser Asp Ser Gly Thr Ser 130 135 140 Ser Gly Leu Leu Pro Ser Asp Arg Val Leu Trp Ile Gly Asp Val Ala 145 150 155 160 Cys Gln Pro Met Thr Pro Ile Pro Glu Glu Thr Phe Leu Glu Leu Lys 165 170 175 Ser Phe Ser Gln Ser Glu Phe Pro Asp Ile Cys Lys Ile Asp Gly Ile 180 185 190 Val Phe Asn Gln Cys Glu Gly Glu Ser Leu Pro Gln Pro Phe Asp Val 195 200 205 Ala Trp Met Asp Val Gly His Ser His Lys Ile Ile Met Arg Glu His 210 215 220 Lys Thr Lys Trp Val Gln Glu Ser Ser Ser Lys Asp Phe Val Cys Tyr 225 230 235 240 Lys Glu Gly Thr Gly Pro Cys Ser Glu Ser Glu Glu Lys Thr Cys Lys 245 250 255 Thr Ser Gly Ser Cys Arg Gly Asp Met Gln Phe Cys Lys Val Ala Gly 260 265 270 Cys Glu His Gly Glu Glu Ala Ser Glu Ala Lys Cys Arg Cys Ser Leu 275 280 285 Val His Lys Pro Gly Glu Val Val Val Ser Tyr Gly Gly Met Arg Val 290 295 300 Arg Pro Lys Cys Tyr Gly Phe Ser Arg Met Met Ala Thr Leu Glu Val 305 310 315 320 Asn Gln Pro Glu Gln Arg Ile Gly Gln Cys Thr Gly Cys His Leu Glu 325 330 335 Cys Ile Asn Gly Gly Val Arg Leu Ile Thr Leu Thr Ser Glu Leu Lys 340 345 350 Ser Ala Thr Val Cys Ala Ser His Phe Cys Ser Ser Ala Thr Ser Gly 355 360 365 Lys Lys Ser Thr Glu Ile Gln Phe His Ser Gly Ser Leu Val Gly Lys 370 375 380 Thr Ala Ile His Val Lys Gly Ala Leu Val Asp Gly Thr Glu Phe Thr 385 390 395 400 Phe Glu Gly Ser Cys Met Phe Pro Asp Gly Cys Asp Ala Val Asp Cys 405 410 415 Thr Phe Cys Arg Glu Phe Leu Lys Asn Pro Gln Cys Tyr Pro Ala Lys 420 425 430 Lys Gly Gln Ala Gly Gln 435 <210> 2 <211> 473 <212> PRT <213> Unknown(未知) <220> <223> SFTS virus Gc protein extracellular domain <400> 2 Cys Asp Glu Met Val His Ala Asp Ser Lys Leu Val Ser Cys Arg Gln 1 5 10 15 Gly Ser Gly Asn Met Lys Glu Cys Val Thr Thr Gly Arg Ala Leu Leu 20 25 30 Pro Ala Val Asn Pro Gly Gln Glu Ala Cys Leu His Phe Thr Ala Pro 35 40 45 Gly Ser Pro Asp Ser Lys Cys Leu Lys Ile Lys Val Lys Arg Ile Asn 50 55 60 Leu Lys Cys Lys Lys Ser Ser Ser Tyr Phe Val Pro Asp Ala Arg Ser 65 70 75 80 Arg Cys Thr Ser Val Arg Arg Cys Arg Trp Ala Gly Asp Cys Gln Ser 85 90 95 Gly Cys Pro Pro His Phe Thr Ser Asn Ser Phe Ser Asp Asp Trp Ala 100 105 110 Gly Lys Met Asp Arg Ala Gly Leu Gly Phe Ser Gly Cys Ser Asp Gly 115 120 125 Cys Gly Gly Ala Ala Cys Gly Cys Phe Asn Ala Ala Pro Ser Cys Ile 130 135 140 Phe Trp Arg Lys Trp Val Glu Asn Pro His Gly Ile Ile Trp Lys Val 145 150 155 160 Ser Pro Cys Ala Ala Trp Val Pro Ser Ala Val Ile Glu Leu Thr Met 165 170 175 Pro Ser Gly Glu Val Arg Thr Phe His Pro Met Ser Gly Ile Pro Thr 180 185 190 Gln Val Phe Lys Gly Val Ser Val Thr Tyr Leu Gly Ser Asp Met Glu 195 200 205 Val Ser Gly Leu Thr Asp Leu Cys Glu Ile Glu Glu Leu Lys Ser Lys 210 215 220 Lys Leu Ala Leu Ala Pro Cys Asn Gln Ala Gly Met Gly Val Val Gly 225 230 235 240 Lys Val Gly Glu Ile Gln Cys Ser Ser Glu Glu Ser Ala Arg Thr Ile 245 250 255 Lys Lys Asp Gly Cys Ile Trp Asn Ala Asp Leu Val Gly Ile Glu Leu 260 265 270 Arg Val Asp Asp Ala Val Cys Tyr Ser Lys Ile Thr Ser Val Glu Ala 275 280 285 Val Ala Asn Tyr Ser Ala Ile Pro Thr Thr Ile Gly Gly Leu Arg Phe 290 295 300 Glu Arg Ser His Asp Ser Gln Gly Lys Ile Ser Gly Ser Pro Leu Asp 305 310 315 320 Ile Thr Ala Ile Arg Gly Ser Phe Ser Val Asn Tyr Arg Gly Leu Arg 325 330 335 Leu Ser Leu Ser Glu Ile Thr Ala Thr Cys Thr Gly Glu Val Thr Asn 340 345 350 Val Ser Gly Cys Tyr Ser Cys Met Thr Gly Ala Lys Val Ser Ile Lys 355 360 365 Leu His Ser Ser Lys Asn Ser Thr Ala His Val Arg Cys Lys Gly Asp 370 375 380 Glu Thr Ala Phe Ser Val Leu Glu Gly Val His Ser Tyr Thr Val Ser 385 390 395 400 Leu Ser Phe Asp His Ala Val Val Asp Glu Gln Cys Gln Leu Asn Cys 405 410 415 Gly Gly His Glu Ser Gln Val Thr Leu Lys Gly Asn Leu Ile Phe Leu 420 425 430 Asp Val Pro Lys Phe Val Asp Gly Ser Tyr Met Gln Thr Tyr His Ser 435 440 445 Thr Val Pro Thr Gly Ala Asn Ile Pro Ser Pro Thr Asp Trp Leu Asn 450 455 460 Ala Leu Phe Gly Asn Gly Leu Ser Arg 465 470 <210> 3 <211> 245 <212> PRT <213> Unknown(未知) <220> <223> NP protein of SFTS virus <400> 3 Met Ser Glu Trp Ser Arg Ile Ala Val Glu Phe Gly Glu Gln Gln Leu 1 5 10 15 Asn Leu Ser Glu Leu Glu Asp Phe Ala Arg Glu Leu Ala Tyr Glu Gly 20 25 30 Leu Asp Pro Ala Leu Ile Ile Lys Lys Leu Lys Glu Thr Gly Gly Asp 35 40 45 Asp Trp Val Lys Asp Thr Lys Phe Ile Ile Val Phe Ala Leu Thr Arg 50 55 60 Gly Asn Lys Ile Val Lys Ala Ser Gly Lys Met Ser Asn Ser Gly Ser 65 70 75 80 Lys Arg Leu Met Ala Leu Gln Glu Lys Tyr Gly Leu Val Glu Arg Ala 85 90 95 Glu Thr Arg Leu Ser Ile Thr Pro Val Arg Val Ala Gln Ser Leu Pro 100 105 110 Thr Trp Thr Cys Ala Ala Ala Ala Ala Leu Lys Glu Tyr Leu Pro Val 115 120 125 Gly Pro Ala Val Met Asn Leu Lys Val Glu Asn Tyr Pro Pro Glu Met 130 135 140 Met Cys Met Ala Phe Gly Ser Leu Ile Pro Thr Ala Gly Val Ser Glu 145 150 155 160 Ala Thr Thr Lys Thr Leu Met Glu Ala Tyr Ser Leu Trp Gln Asp Ala 165 170 175 Phe Thr Lys Thr Ile Asn Val Lys Met Arg Gly Ala Ser Lys Thr Glu 180 185 190 Val Tyr Asn Ser Phe Arg Asp Pro Leu His Ala Ala Val Asn Ser Val 195 200 205 Phe Phe Pro Asn Asp Val Arg Val Lys Trp Leu Lys Ala Lys Gly Ile 210 215 220 Leu Gly Pro Asp Gly Val Pro Ser Arg Ala Ala Glu Val Ala Ala Ala 225 230 235 240 A La Tyr Arg Asn Leu 245 <210> 4 <211> 1299 <212> DNA <213> Unknown <220> <223> Gene encoding the extracellular domain of the Gn protein of SFTS virus <400> 4 gactctggac ctattatctg tgctggacct attcattcaa acaaatctgc tgacatccct 60 catctgctgg gctactccga aaaaatctgc cagattgatc gactgatcca cgtgagctcc 120 tggctgcgga accatagcca gttccaggga tacgtcggac agaggggcgg gcgctctcag 180 gtgagttat atccagccga gaacagctac agccggtggt ccggactgct gtctccatgt 240 gacgcagatt ggctgggaat gctggtggtc aagaaagcta agggctcaga tatgattgtc 300 cccggcccta gctacaaggg gaaagtgttc tttgaacgcc ccaccttcga cggctatgtg 360 ggctgggggt gcggatcagg caaaagccga acagagtccg gagaactgtg cagcagcgac 420 tctggaactt caagcggcct gctgcctagc gatagggtcc tgtggattgg cgacgtggcc 480 tgccagccaa tgaccccaat ccccgaggaa acatttctgg agctgaagag tttctcacag 540 agcgaatttc cagacatctg caaaattgac ggcatcgtgt tcaaccagtg tgagggggaa 600 agcctgcctc agccatttga tgtcgcttgg atggacgtgg gccactccca taagatcatt 660 atgcgcgagc acaagactaa atgggtccag gaatcctcta gtaaggattt cgtgtgctac 720 aaagagggga ccggaccctg ttccgaatct gaggaaaaga cttgcaaaac cagtgggtca 780 tgtcggggag acatgcagtt ttgcaaggtg gctggctgtg agcacgggga ggaagcctct 840 gaagctaaat gcagatgtag tctggtccat aaacccggcg aggtggtcgt gagttacgga 900 ggcatgcggg tgagacctaa atgctatggg ttcagcagaa tgatggcaac cctggaggtg 960 aaccagcctg aacagaggat cggacagtgc acaggctgtc acctggagtg tattaatggg 1020 ggagtccgac tgatcaccct gacaagtgaa ctgaagtcag caacagtgtg cgcctctcat 1080 ttctgttcaa gcgccactag cggcaagaaa tccaccgaga ttcagtttca cagcgggtcc 1140 ctggtgggaa agactgctat ccatgtcaaa ggagcactgg tggatggcac agagttcact 1200 tttgaagggt cctgcatgtt tccagacgga tgtgatgcag tggactgcac attctgtcgc 1260 gagtttctga agaatcccca gtgctatcct gccaagaaa 1299 <210> 5 <211> 1419 <212> DNA <213> Unknown(未知) <220> <223> Gene encoding the extracellular domain of the Gc protein of the SFTS virus <400> 5 tgcgatgaaa tggtccacgc cgatagcaaa ctggtcagct gtaggcaggg aagcgggaac 60 atgaaggaat gcgtcacaac cggaagagct ctgctgccag cagtgaaccc tggacaggag 120 gcctgcctgc acttcacagc tcctggaagt ccagactcaa aatgcctgaa gatcaaagtg 180 aagcggatta atctgaagtg taagaaaagc tcctcttact ttgtgcccga cgcaaggtca 240 cgctgcacta gcgtccggag atgtagatgg gcaggcgatt gccagagcgg atgtccacct 300 catttcacct ctaacagttt ttcagacgat tgggcaggca agatggacag ggcaggactg 360 gggttcagcg gatgctccga tggatgtgga ggagcagctt gcggatgttt caacgcagcc 420 ccctcctgca tcttttggcg caaatgggtg gagaatcctc acggcatcat ttggaaggtc 480 tccccttgtg ctgcatgggt gccatctgcc gtcatcgagc tgacaatgcc cagcggcgaa 540 gtgagaactt tccatccaat gtccgggatt cccacccagg tctttaaggg agtgtctgtc 600 acatatctgg gcagcgacat ggaggtgtcc gggctgactg atctgtgcga aatcgaggaa 660 ctgaaatcca agaaactggc cctggctcct tgtaatcagg caggaatggg agtggtcggc 720 aaggtcgggg agattcagtg cagttcagag gaaagcgcac gcaccatcaa gaaagacggg 780 tgtatttgga acgccgatct ggtgggaatc gagctgcgag tggacgatgc tgtctgctac 840 tccaaaatta catctgtgga agcagtcgcc aattattccg ctatccctac cacaattggc 900 ggcctgcgat tcgaacggtc tcacgacagt caggggaaga tcagcggcag ccccctggac 960 atcaccgcca ttcgaggatc ttttagtgtg aactacagag gcctgaggct gtcactgagc 1020 gagatcactg ctacctgcac aggagaagtg actaatgtca gtggctgcta ttcatgtatg 1080 accggggcaa aagtgagtat taagctgcac agctccaaaa actcaacagc tcatgtgcgc 1140 tgtaagggag acgagactgc attctctgtg ctggaaggcg tccacagtta cacagtgtcc 1200 ctgtcttttg accatgccgt ggtcgatgag cagtgccagc tgaactgtgg gggacacgaa 1260 agccaagtga ctctgaaagg caatctgatc ttcctggacg tgccaaagtt tgtcgatggg 1320 tcatacatgc agacctatca tagcactgtg cccaccggcg ccaatattcc atctcccaca 1380 gattggctga acgctctgtt tgggaatgga ctgagtcgg 1419 <210> 6 <211> 60 <212> DNA <213> Unknown(未知) <220> <223> Gene encoding signal peptide <400> 6 atggagacag acacactcct gctatgggta ctgctgctct gggttccagg ttccactggt 60 <210> 7 <211> 654 <212> DNA <213> Unknown(未知) <220> Gene encoding human Fc <400> 7 gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc 60 ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac 120 cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag 180 ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac 240 caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc 300 cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca ggtgtacacc 360 ctgcccccat cccgggatga gctgaccaag aaccaggtca gcctgacctg cctggtcaaa 420 ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac 480 tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaagctc 540 accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag 600 gctctgcaca accactacac gcagaagagc ctctccctgt ccccgggtaa atga 654 <210> 8 <211> 60 <212> DNA <213> Unknown <220> <223> Gene encoding hinge region <400> 8 ggccaggccg gccaggagcc caaatctagc gacaaaactc acacaagccc accgagccca 60 <210> 9 <211> 738 <212> DNA <213> Unknown <220> <223> Gene encoding SFTS virus NP protein <400> 9 atgtcggagt ggtccaggat tgcagtggag tttggtgagc agcagctcaa tttgtctgag 60 cttgaggatt tcgcgagaga actggcctat gaaggccttg accctgcttt gatcatcaag 120 aagctgaagg agacaggtgg agatgattgg gtgaaggata caaagttcat cattgtcttt 180 gccctgactc gaggcaacaa gatcgtcaag gcatcaggga aaatgtcaaa ctcagggtcc 240 aagaggttga tggcactcca agagaaatat ggactggttg agagggcaga gaccaggctc 300 tcaatcactc ctgtgagggt tgcgcagagc cttcccactt ggacatgtgc tgcagcagca 360 gccttgaagg agtatctccc tgtggggcca gccgtcatga acctgaaggt cgagaattat 420 ccccctgaga tgatgtgcat ggcctttgga tccctgattc caactgcggg ggtatctgaa 480 gccacaacca agaccctgat ggaggcctac tctctgtggc aagatgcctt caccaagact 540 atcaatgtga agatgcgcgg agccagcaag acagaagtttt acaacctcctt cagggaccct 600 cttcatgctg ctgtgaactc tgtctcttt cccaatgatg ttcgggtaaa gtggctgaag 660 gccaagggga tccttggccc agatggggtc cccagcagag ctgctgaggt tgctgctgct 720 gcttacagaa acctgtaa 738 <210> 10 <211> 676 <212> PRT <213> Unknown <220> <223> Gn-Fc <400> 10 Asp Val Ala Gln Ala Ala Asp Ser Gly Pro Ile Ile Cys Ala Gly Pro 1 5 10 15 His Ser Ser Asn Lys Ser Ala Asp Ile Pro His Leu Leu Gly Tyr Ser 20 25 30 Glu Lys Ile Cys Gln Ile Asp Arg Leu Ile His Will Be Trp Leu 35 40 45 Arg Asn His Is Gln Phe Gln Gly Tyr Val Gly Gln Arg Gly Gly Arg 50 55 60 Ser Gln Val Ser Tyr Tyr Pro Ala Glu Asn Ser Tyr Ser Arg Trp Ser 65 70 75 80 Gly Leu Leu Ser Pro Cys Asp Ala Asp Trp Leu Gly Met Leu Val Val 85 90 95 Lys Lys Ala Lys Gly Ser Asp Met Ile Val Pro Gly Pro Ser Tyr Lys 100 105 110 Gly Lys Val Phe Phe Glu Arg Pro Thr Phe Asp Gly Tyr Val Gly Trp 115 120 125 Gly Cys Gly Ser Gly Lys Ser Arg Thr Glu Ser Gly Glu Leu Cys Ser 130 135 140 Ser Asp Ser Gly Thr Ser Ser Gly Leu Leu Pro Ser Asp Arg Val Leu 145 150 155 160 Trp Ile Gly Asp Val Ala Cys Gln Pro Met Thr Pro Ile Pro Glu Glu 165 170 175 Thr Phe Leu Glu Leu Lys Ser Phe Ser Gln Ser Glu Phe Pro Asp Ile 180 185 190 Cys Lys Ile Asp Gly Ile Val Phe Asn Gln Cys Glu Gly Glu Ser Leu 195 200 205 Pro Gln Pro Phe Asp Val Ala Trp Met Asp Val Gly His Ser His Lys 210 215 220 Ile Ile Met Arg Glu His Lys Thr Lys Trp Val Gln Glu Ser Ser Ser 225 230 235 240 Lys Asp Phe Val Cys Tyr Lys Glu Gly Thr Gly Pro Cys Ser Glu Ser 245 250 255 Glu Glu Lys Thr Cys Lys Thr Ser Gly Ser Cys Arg Gly Asp Met Gln 260 265 270 Phe Cys Lys Val Ala Gly Cys Glu His Gly Glu Glu Ala Ser Glu Ala 275 280 285 Lys Cys Arg Cys Ser Leu Val His Lys Pro Gly Glu Val Val Val Ser 290 295 300 Tyr Gly Gly Met Arg Val Arg Pro Lys Cys Tyr Gly Phe Ser Arg Met 305 310 315 320 Met Ala Thr Leu Glu Val Asn Gln Pro Glu Gln Arg Ile Gly Gln Cys 325 330 335 Thr Gly Cys His Leu Glu Cys Ile Asn Gly Gly Val Arg Leu Ile Thr 340 345 350 Leu Thr Ser Glu Leu Lys Ser Ala Thr Val Cys Ala Ser His Phe Cys 355 360 365 Ser Ser Ala Thr Ser Gly Lys Lys Ser Thr Glu Ile Gln Phe His Ser 370 375 380 Gly Ser Leu Val Gly Lys Thr Ala Ile His Val Lys Gly Ala Leu Val 385 390 395 400 Asp Gly Thr Glu Phe Thr Phe Glu Gly Ser Cys Met Phe Pro Asp Gly 405 410 415 Cys Asp Ala Val Asp Cys Thr Phe Cys Arg Glu Phe Leu Lys Asn Pro 420 425 430 Gln Cys Tyr Pro Ala Lys Lys Gly Gln Ala Gly Gln Glu Pro Lys Ser 435 440 445 Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu 450 455 460 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 465 470 475 480 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 485 490 495 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 500 505 510 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 515 520 525 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 530 535 540 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 545 550 555 560 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 565 570 575 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 580 585 590 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 595 600 605 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 610 615 620 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 625 630 635 640 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 645 650 655 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 660 665 670 Ser Pro Gly Lys 675 <210> 11 <211> 711 <212> PRT <213> Unknown(未知) <220> <223> Gc-Fc <400> 11 Asp Val Ala Gln Ala Ala Cys Asp Glu Met Val His Ala Asp Ser Lys 1 5 10 15 Leu Val Ser Cys Arg Gln Gly Ser Gly Asn Met Lys Glu Cys Val Thr 20 25 30 Thr Gly Arg Ala Leu Leu Pro Ala Val Asn Pro Gly Gln Glu Ala Cys 35 40 45 Leu His Phe Thr Ala Pro Gly Ser Pro Asp Ser Lys Cys Leu Lys Ile 50 55 60 Lys Val Lys Arg Ile Asn Leu Lys Cys Lys Lys Ser Ser Ser Tyr Phe 65 70 75 80 Val Pro Asp Ala Arg Ser Arg Cys Thr Ser Val Arg Arg Cys Arg Trp 85 90 95 Ala Gly Asp Cys Gln Ser Gly Cys Pro Pro His Phe Thr Ser Asn Ser 100 105 110 Phe Ser Asp Asp Trp Ala Gly Lys Met Asp Arg Ala Gly Leu Gly Phe 115 120 125 Ser Gly Cys Ser Asp Gly Cys Gly Gly Ala Ala Cys Gly Cys Phe Asn 130 135 140 Ala Ala Pro Ser Cys Ile Phe Trp Arg Lys Trp Val Glu Asn Pro His 145 150 155 160 Gly Ile Ile Trp Lys Val Ser Pro Cys Ala Ala Trp Val Pro Ser Ala 165 170 175 Val Ile Glu Leu Thr Met Pro Ser Gly Glu Val Arg Thr Phe His Pro 180 185 190 Met Ser Gly Ile Pro Thr Gln Val Phe Lys Gly Val Ser Val Thr Tyr 195 200 205 Leu Gly Ser Asp Met Glu Val Ser Gly Leu Thr Asp Leu Cys Glu Ile 210 215 220 Glu Glu Leu Lys Ser Lys Lys Leu Ala Leu Ala Pro Cys Asn Gln Ala 225 230 235 240 Gly Met Gly Val Val Gly Lys Val Gly Glu Ile Gln Cys Ser Ser Glu 245 250 255 Glu Ser Ala Arg Thr Ile Lys Lys Asp Gly Cys Ile Trp Asn Ala Asp 260 265 270 Leu Val Gly Ile Glu Leu Arg Val Asp Asp Ala Val Cys Tyr Ser Lys 275 280 285 Ile Thr Ser Val Glu Ala Val Ala Asn Tyr Ser Ala Ile Pro Thr Thr 290 295 300 Ile Gly Gly Leu Arg Phe Glu Arg Ser His Asp Ser Gln Gly Lys Ile 305 310 315 320 Ser Gly Ser Pro Leu Asp Ile Thr Ala Ile Arg Gly Ser Phe Ser Val 325 330 335 Asn Tyr Arg Gly Leu Arg Leu Ser Leu Ser Glu Ile Thr Ala Thr Cys 340 345 350 Thr Gly Glu Val Thr Asn Val Ser Gly Cys Tyr Ser Cys Met Thr Gly 355 360 365 Ala Lys Val Ser Ile Lys Leu His Ser Ser Lys Asn Ser Thr Ala His 370 375 380 Val Arg Cys Lys Gly Asp Glu Thr Ala Phe Ser Val Leu Glu Gly Val 385 390 395 400 His Ser Tyr Thr Val Ser Leu Ser Phe Asp His Ala Val Val Asp Glu 405 410 415 Gln Cys Gln Leu Asn Cys Gly Gly His Glu Ser Gln Val Thr Leu Lys 420 425 430 Gly Asn Leu Ile Phe Leu Asp Val Pro Lys Phe Val Asp Gly Ser Tyr 435 440 445 Met Gln Thr Tyr His Ser Thr Val Pro Thr Gly Ala Asn Ile Pro Ser 450 455 460 Pro Thr Asp Trp Leu Asn Ala Leu Phe Gly Asn Gly Leu Ser Arg Glu 465 470 475 480 Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro 485 490 495 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 500 505 510 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 515 520 525 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 530 535 540 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 545 550 555 560 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 565 570 575 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 580 585 590 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 595 600 605 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 610 615 620 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 625 630 635 640 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 645 650 655 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 660 665 670 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 675 680 685 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 690 695 700 Leu Ser Leu Ser Pro Gly Lys 705 710 <210> 12 <211> 245 <212> PRT <213> Unknown(未知) <220> <223> NP <400> 12 Met Ser Glu Trp Ser Arg Ile Ala Val Glu Phe Gly Glu Gln Gln Leu 1 5 10 15 Asn Leu Ser Glu Leu Glu Asp Phe Ala Arg Glu Leu Ala Tyr Glu Gly 20 25 30 Leu Asp Pro Ala Leu Ile Ile Lys Lys Leu Lys Glu Thr Gly Gly Asp 35 40 45 Asp Trp Val Lys Asp Thr Lys Phe Ile Ile Val Phe Ala Leu Thr Arg 50 55 60 Gly Asn Lys Ile Val Lys Ala Ser Gly Lys Met Ser Asn Ser Gly Ser 65 70 75 80 Lys Arg Leu Met Ala Leu Gln Glu Lys Tyr Gly Leu Val Glu Arg Ala 85 90 95 Glu Thr Arg Leu Ser Ile Thr Pro Val Arg Val Ala Gln Ser Leu Pro 100 105 110 Thr Trp Thr Cys Ala Ala Ala Ala Ala Leu Lys Glu Tyr Leu Pro Val 115 120 125 Gly Pro Ala Val Met Asn Leu Lys Val Glu Asn Tyr Pro Pro Glu Met 130 135 140 Met Cys Met Ala Phe Gly Ser Leu Ile Pro Thr Ala Gly Val Ser Glu 145 150 155 160 Ala Thr Thr Lys Thr Leu Met Glu Ala Tyr Ser Leu Trp Gln Asp Ala 165 170 175 Phe Thr Lys Thr Ile Asn Val Lys Met Arg Gly Ala Ser Lys Thr Glu 180 185 190 Val Tyr Asn Ser Phe Arg Asp Pro Leu His Ala Ala Val Asn Ser Val 195 200 205 Phe Phe Pro Asn Asp Val Arg Val Lys Trp Leu Lys Ala Lys Gly Ile 210 215 220 Leu Gly Pro Asp Gly Val Pro Ser Arg Ala Ala Glu Val Ala Ala Ala 225 230 235 240 Ala Tyr Arg Asn Leu 245

Claims

1. A vaccine composition for fever with thrombocytopenia syndrome virus, characterized in that: The active ingredient is a mixture of the extracellular domain of the Gc protein fused to the Fc site of a human antibody and the full-length NP protein, or a mixture of the extracellular domain of the Gn protein fused to the Fc site of a human antibody and the full-length NP protein. The extracellular domain of the Gc protein fused to the Fc site of the human antibody is composed of the amino acid sequence of SEQ ID NO: 11 excluding the N-terminal AspValAlaGlnAlaAla; The extracellular domain of the Gn protein fused to the Fc site of the human antibody is composed of the amino acid sequence of SEQ ID NO: 10 excluding the N-terminal AspValAlaGlnAlaAla; The full-length NP protein consists of the amino acid sequence of sequence number 3.

2. The vaccine composition according to claim 1, characterized in that: The composition comprises a pharmaceutically acceptable carrier.

3. The vaccine composition according to claim 2, characterized in that: The pharmaceutically acceptable carrier comprises a diluent.

4. The vaccine composition according to claim 2, characterized in that: The pharmaceutically acceptable carrier comprises an excipient.

5. The vaccine composition according to claim 2, characterized in that: The pharmaceutically acceptable carrier comprises a stabilizer.

6. The vaccine composition according to claim 2, characterized in that: The pharmaceutically acceptable carrier contains a preservative.

7. The vaccine composition according to claim 3, characterized in that: The diluent is propylene glycol, polyethylene glycol or vegetable oil.

8. The vaccine composition according to claim 4, characterized in that: The excipient is starch, glucose, lactose, sucrose, gelatin, malt, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talcum powder, anhydrous skim milk, glycerol, propylene, ethylene glycol or ethanol.

9. The vaccine composition according to claim 5, characterized in that: The stabilizer is sorbitol, mannitol, dextran or glutamic acid.

10. The vaccine composition according to claim 6, characterized in that: The preservative is thimerosal, thimerosal, gentamicin, neomycin, nystatin, amphotericin B, tetracycline, penicillin, streptomycin or polymyxin B.

11. The vaccine composition according to claim 1, characterized in that: The composition also comprises an adjuvant.

12. The vaccine composition according to claim 11, characterized in that: The adjuvant is an aluminum salt in gel form.

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