Patent application title: AVIPOX RECOMBINANTS EXPRESSING FOOT AND MOUTH DISEASE VIRUS GENES
Inventors:
Robert Nordgren (Athens, GA, US)
Sheena May Loosmore (Aurora, CA)
Jean Christophe Francis Audonnet (Lyon, FR)
Marvin J. Grubman (Southold, NY, US)
IPC8 Class: AC12N1566FI
USPC Class:
435 9141
Class name: Polynucleotide (e.g., nucleic acid, oligonucleotide, etc.) modification or preparation of a recombinant dna vector by insertion or addition of one or more nucleotides
Publication date: 2009-10-08
Patent application number: 20090253185
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Patent application title: AVIPOX RECOMBINANTS EXPRESSING FOOT AND MOUTH DISEASE VIRUS GENES
Inventors:
Robert Nordgren
Sheena May Loosmore
Jean-Christophe Francis Audonnet
Marvin J. Grubman
Agents:
FROMMER LAWRENCE & HAUG
Assignees:
Origin: NEW YORK, NY US
IPC8 Class: AC12N1566FI
USPC Class:
435 9141
Patent application number: 20090253185
Abstract:
The present invention relates to modified poxyiral vectors and to methods
of making and using the same. In particular, the invention relates to
recombinant avipox that expresses gene products of foot and mouth disease
virus (FMDV), and to compositions or vaccines that elicit immune
responses directed to FMDV gene products and which can confer protective
immunity against infection by FMDV.Claims:
1-30. (canceled)
31. A method of producing a recombinant avipox vector comprising at least one nucleic acid molecule encoding one or more foot-and-mouth disease virus (FMDV) antigen(s), comprising the steps of:(a) linearizing a donor plasmid with a restriction endonuclease, wherein the donor plasmid comprises restriction endonuclease cleavage sites or a multiple cloning site; and(b) ligating at least one nucleic acid molecule comprising(i) a nucleic acid sequence encoding one or more FMDV antigen(s),(ii) a viral promoter sequence, and(iii) insertion sequences flanking (i) and (ii) that have complementary restriction endonuclease cleavage sites to the donor plasmid at FMDV antigens,thereby producing the recombinant avipox vector.
32. The method of claim 31, further comprising the steps of:(c) introducing the vector into a cell permissive for replication of the vector; and(d) isolating the vector from the cell.
33. The method of claim 31, wherein the avipox is ALVAC.
34. The method of claim 31, wherein the avipox is fowlpox.
35. The method of claim 31, wherein the antigen comprises at least one of FMDV VP1, VP2, VP3, VP4, 2A, 2B, and 3C.
36. The method of claim 31, wherein the nucleic acid sequence encoding one or more FMDV antigen(s) is a cDNA encoding FMDV P1 region and a cDNA encoding FMDV 3C protease.
37. The method of claim 31, wherein the promoter sequence is selected from the group consisting of H6 vaccinia promoter, I3L vaccinia promoter, 42K poxyiral promoter, 7.5K vaccinia promoter and Pi vaccinia promoter.
38. The method of claim 37, wherein the promoter is the H6 vaccinia promoter, which is mutated such that expression levels of the FMDV antigens are decreased compared with expression levels of the FMDV antigens under a wild type H6 vaccinia promoter.
39. The method of claim 31, wherein the vector comprises a C6 insertion locus, and wherein flanking sequences of the C6 insertion locus promote homologous recombination of the FMDV antigens with the C6 insertion locus.
40. The method of claim 39, wherein the flanking sequences comprise C6L and C6R open reading frames of avipox.
41. The method of claim 31, wherein the vector comprises a F8 insertion locus, and wherein flanking sequences of the F8 insertion locus promote homologous recombination of the FMDV antigens with the F8 insertion locus.
42. The method of claim 41, wherein the flanking sequences comprise F8L and F8R open reading frames of avipox.
43. The method of claim 31, wherein the vector further comprises a reporter gene.
44. The method of claim 43, wherein the reporter gene is selected from the group consisting of neomycin resistance gene, ampicillin resistance gene, lacZ (quadrature-galactosidase), luciferase, and green fluorescent protein (GFP).
45. The method of claim 32, wherein the cell permissive for growth of the vector is a chicken embryonic fibroblast.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001]This application is a divisional of U.S. application Ser. No. 11/110,480, filed Apr. 20, 2005, which claims priority to provisional U.S. application Ser. No. 60/563,786 filed on Apr. 20, 2004.
[0002]This application makes reference to U.S. application Ser. No. 10/327,481, filed on Dec. 20, 2002, which is a continuation of International application No. PCT/FR01/02042, filed on Jun. 27, 2001, published on Jan. 3, 2002 as WO 02/00251, and claiming priority to French application No. 00/08437, filed on Jun. 29, 2000.
[0003]All of the foregoing applications, as well as all documents cited in the foregoing applications ("application documents") and all documents cited or referenced in the application documents are incorporated herein by reference. Also, all documents cited in this application ("herein-cited documents") and all documents cited or referenced in herein-cited documents are incorporated herein by reference. In addition, any manufacturer's instructions or catalogues for any products cited or mentioned in each of the application documents or herein-cited documents are incorporated by reference. Documents incorporated by reference into this text or any teachings therein can be used in the practice of the invention. Documents incorporated by reference into this text are not admitted to be prior art.
FIELD OF THE INVENTION
[0004]The present invention relates to vectors, such as viruses, e.g., modified viruses such as poxviruses, and to methods of making and using the same. In particular, the invention relates to recombinant avipox vectors and viruses that express antigens of foot and mouth disease virus (FMDV), and to methods of making and using the same. The invention further relates to methods of eliciting an immune response to FMDV in a subject.
BACKGROUND OF THE INVENTION
[0005]Foot-and-mouth disease (FMD) is one of the most virulent and contagious diseases affecting farm animals. This disease is endemic in numerous countries in the world, especially in Africa, Asia and South America. In addition, epidemic outbreaks can occur periodically. The presence of this disease in a country may have very severe economic consequences resulting from loss of productivity, loss of weight and milk production in infected herds, and from trade embargoes imposed on these countries. The measures taken against this disease consist of strict application of import restrictions, hygiene controls and quarantine, slaughtering sick animals and vaccination programs using inactivated vaccines, either as a preventive measure at the national or regional level, or periodically when an epidemic outbreak occurs.
[0006]FMD is characterized by its short incubation period, its highly contagious nature, the formation of ulcers in the mouth and on the feet and sometimes, the death of young animals. FMD affects a number of animal species, in particular cattle, pigs, sheep and goats. The agent responsible for this disease is a ribonucleic acid (RNA) virus belonging to the Aphthovirus genus of the Picornaviridae family (Cooper et al., Intervirology, 1978, 10, 165-180). At present, at least seven types of foot-and-mouth disease virus (FMDV) are known: the European types (A, O and C), the African types (SAT1, SAT2 and SAT3) and an Asiatic type (Asia 1). Numerous sub-types have also been distinguished (Kleid et al. Science (1981), 214, 1125-1129).
[0007]FMDV is a naked icosahedral virus of about 25 nm in diameter, containing a single-stranded RNA molecule consisting of about 8500 nucleotides, with a positive polarity. This RNA molecule comprises a single open reading frame (ORF), encoding a single polyprotein containing, inter alia, the capsid precursor also known as protein P1 or P88. The protein P1 is myristylated at its amino-terminal end. During the maturation process, the protein P1 is cleaved by the protease 3C into three proteins known as VP0, VP1 and VP3 (or 1AB, 1D and 1C respectively; Belsham G. J., Progress in Biophysics and Molecular Biology, 1993, 60, 241-261). In the virion, the protein VP0 is then cleaved into two proteins, VP4 and VP2 (or 1A and 1B respectively). The mechanism for the conversion of the proteins VP0 into VP 1 and VP3, and for the formation of mature virions is not known. The proteins VP1, VP2 and VP3 have a molecular weight of about 26,000 Da, while the protein VP4 is smaller at about 8,000 Da.
[0008]The simple combination of the capsid proteins forms the protomer or 5S molecule, which is the elementary constituent of the FMDV capsid. This protomer is then complexed into a pentamer to form the 12S molecule. The virion results from the encapsidation of a genomic RNA molecule by assembly of twelve 12S pentamers, thus constituting the 146S particles. The viral capsid may also be formed without the presence of an RNA molecule inside it (hereinafter "empty capsid"). The empty capsid is also designated as particle 70S. The formation of empty capsids may occur naturally during viral replication or may be produced artificially by chemical treatment.
[0009]Many hypotheses, research routes, and proposals have been developed in an attempt to design effective vaccines against FMD. Currently, the only vaccines on the market comprise inactivated virus. Concerns about safety of the FMDV vaccine exist, as outbreaks of FMD in Europe have been associated with shortcomings in vaccine manufacture (King, A. M. Q. et al, (1981) Nature 293: 479-480). The inactivated vaccines do not confer long-term immunity, thus requiring booster injections given every year, or more often in the event of epidemic outbreaks. In addition, there are risks linked to incomplete inactivation and/or to the escape of virus during the production of inactivated vaccines (King, A. M. Q., ibid). A goal in the art has been to construct conformationally correct immunogens lacking the infective FMDV genome to make effective and safe vaccines.
[0010]Vaccinia virus has been used successfully to immunize against smallpox, culminating in the worldwide eradication of smallpox in 1980. Thus, a new role for poxviruses became important, that of a genetically engineered vector for the expression of foreign genes (Panicali and Paoletti, 1982; Paoletti et al., 1984). Genes encoding heterologous antigens have been expressed in vaccinia, often resulting in protective immunity against challenge by the corresponding pathogen (reviewed in Tartaglia et al., 1990). A highly attenuated strain of vaccines, designated MVA, has also been used as a vector for poxvirus-based vaccines. Use of MVA is described in U.S. Pat. No. 5,185,146.
[0011]Additional vaccine vector systems involve the use of avipox viruses, which are naturally host-restricted poxviruses. Both fowlpoxvirus (FPV; Taylor et al. 1988a, b) and canarypoxvirus (CPV; Taylor et al., 1991 & 1992) have been engineered to express foreign gene products. Fowlpox virus (FPV) is the prototypic virus of the Avipox genus of the Poxvirus family. The virus causes an economically important disease of poultry that has been well controlled since the 1920's by the use of live attenuated vaccines. Replication of the avipox viruses is limited to avian species (Matthews, 1982) and there are no reports in the literature of avipox virus causing a productive infection in any non-avian species including man. This host restriction provides an inherent safety barrier against transmission of the virus to other species and makes the use of avipox virus based vaccine vectors in veterinary and human applications an attractive proposition.
[0012]Other attenuated poxvirus vectors have been prepared by genetic modifications of wild type strains of virus. The NYVAC vector, derived by deletion of specific virulence and host-range genes from the Copenhagen strain of vaccinia (Tartaglia et al., 1992) has proven useful as a recombinant vector in eliciting a protective immune response against an expressed foreign antigen. Another engineered poxvirus vector is ALVAC, derived from canarypox virus (see U.S. Pat. No. 5,756,103). ALVAC does not productively replicate in non-avian hosts, a characteristic thought to improve its safety profile (Taylor et al., 1991 & 1992). ALVAC was deposited under the terms of the Budapest Treaty with the American Type Culture Collection under accession number VR-2547. Yet another engineered poxvirus vector is TROVAC, derived from fowlpox virus (see U.S. Pat. No. 5,766,599).
[0013]Recombinant poxviruses can be constructed in two steps known in the art and analogous to the methods for creating synthetic recombinants of poxviruses such as the vaccinia virus and avipox virus described in U.S. Pat. Nos. 4,769,330; 4,722,848; 4,603,112; 5,110,587; 5,174,993; 5,494,807; and 5,505,941, the disclosures of which are incorporated herein by reference. It can thus be appreciated that provision of a FMDV recombinant poxvirus, and of compositions and products therefrom, particularly ALVAC or TROVAC-based FMDV recombinants and compositions and products therefrom, especially such recombinants containing the P1 genes and/or C3 protease gene of FMDV, and compositions and products therefrom, would be a highly desirable advance over the current state of technology.
SUMMARY OF THE INVENTION
[0014]Accordingly, one aspect of the present invention provides a recombinant avipox vector comprising at least one nucleic acid molecule encoding one or more foot-and-mouth disease virus (FMDV) antigen(s). In advantageous embodiments, the avipox is ALVAC or TROVAC.
[0015]Advantageously, the FMDV antigen(s) can be VP1, VP2, VP3, VP4, 2A, 2B or 3C. Advantageously, the nucleic acid molecule encoding one or more foot-and-mouth disease virus (FMDV) antigen(s) is a cDNA encoding FMDV P1 region and a cDNA encoding FMDV 3C protease of FMDV.
[0016]In one embodiment, the FMDV antigens are operably linked to a promoter sequence, which can be the H6 vaccinia promoter, I3L vaccinia promoter, 42K vaccinia promoter, 7.5K vaccinia promoter, or Pi vaccinia promoter. In another embodiment, the promoter is the H6 vaccinia promoter, which is mutated such that the expression levels of the FMDV antigens are decreased compared with expression levels of the FMDV antigens under a wild type (i.e. unmutated) H6 vaccinia promoter.
[0017]In another embodiment, the avipox vector of the present invention comprises a C6 insertion locus, wherein flanking sequences of the C6 insertion locus promote homologous recombination of the FMDV antigens with the C6 insertion locus. Advantageously, the flanking sequences comprise the C6L and C6R open reading frames of canarypox.
[0018]In a further embodiment, the avipox vector of the present invention comprises a F8 insertion locus, wherein the flanking sequences of the F8 insertion locus promote homologous recombination of the FMDV antigens with the F8 insertion locus. Advantageously, the flanking sequences comprise the F8L and F8R open reading frames of fowlpox.
[0019]A second aspect of the present invention provides a recombinant avipox virus, comprising at least one nucleic acid molecule encoding one or more FMDV antigens. The present invention also provides recombinant avipox viruses vCP2186, vCP2181, vCP2176, and vFP2215.
[0020]A further aspect of the invention relates to a method of eliciting an immune response to FMDV in a subject, comprising administering the avipox vector or avipox virus of the present invention to the subject.
[0021]In yet another aspect of the present invention, a method of producing a recombinant avipox vector comprising at least one nucleic acid molecule encoding one or more FMDV antigen(s), comprising the steps of: a) linearizing a donor plasmid with a restriction endonuclease, wherein the donor plasmid comprises restriction endonuclease cleavage sites or a multiple cloning site; and b) ligating at least one nucleic acid molecule comprising (i) a nucleic acid sequence encoding one or more FMDV antigen(s), (ii) a viral promoter sequence, and (iii) insertion sequences flanking (i) and (ii) that have complementary restriction endonuclease cleavage sites to the donor plasmid at FMDV antigens, thereby producing the recombinant avipox vector.
[0022]The method can further comprise the steps of c) introducing the vector into a cell permissive for replication of the vector; and d) isolating the vector from the cell. Advantageously, the cell permissive for replication of the vector is a chicken embryonic fibroblast.
[0023]In another embodiment, the vector further comprises a reporter gene, which is selected from the group consisting of the neomycin resistance gene, the ampicillin resistance gene, lacZ (β-galactosidase), luciferase, and green fluorescent protein (GFP).
[0024]These and other objects of the invention will be described in further detail in connection with the detailed description of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0025]The following Detailed Description, given by way of example, but not intended to limit the invention to specific embodiments described, may be understood in conjunction with the accompanying drawings, incorporated herein by reference. Various preferred features and embodiments of the present invention will now be described by way of non-limiting examples and with reference to the accompanying drawings in which:
[0026]FIG. 1 shows the genome of foot and mouth disease virus (FMDV) and the genes inserted into the avipox recombinants.
[0027]FIG. 2 shows the oligonucleotide primers used to PCR-amplify the H6p FMDV gene cassette (SEQ ID NO:1-3), and the amino acids encoded by the nucleotides (SEQ ID NO:4 and 5).
[0028]FIGS. 3A and 3B show the construction of a pC5 H6p FMDV P1+3C donor plasmid for generating ALVAC recombinants, with inserts at the C5 loci.
[0029]FIGS. 4A-4E show the nucleotide (SEQ ID NO:6) and amino acid sequences (SEQ ID NO:7) of the C5 H6p FMDV gene cassette of the pC5 H6p FMDV P1+3C donor plasmid.
[0030]FIGS. 5A and 5B show the construction of a pF8 H6p FMDV P1+3C donor plasmid for generating fowlpox recombinants, with the insert at the unique F8 locus.
[0031]FIGS. 6A-6F show the nucleotide (SEQ ID NO:8) and amino acid sequences (SEQ ID NO:9) of the F8 H6p FMDV gene cassette of the pF8 H6p FMDV P1+3C donor plasmid.
[0032]FIG. 7 shows the oligonucleotide primers used to PCR amplify the 3'-end of the FMDV gene cassette (SEQ ID NO:10-12), and the amino acids encoded by the nucleotides (SEQ ID NO:13 and 14).
[0033]FIG. 8 shows the construction of a promoter-less pC6 FMDV P1+3C insertion plasmid for introduction of different promoters.
[0034]FIGS. 9A and 9B show the construction of a pC6 H6p FMDV P1+3C donor plasmid for generating ALVAC recombinants, with the insert at the unique C6 locus.
[0035]FIGS. 1A-10E show the nucleotide (SEQ ID NO:15) and amino acid sequences (SEQ ID NO:16) of the C6 H6p FMDV gene cassette of the pC6 H6p FMDV P1+3C donor plasmid.
[0036]FIG. 11 shows the nucleotide sequences of the wild-type early/late H6 promoter (H6p) (SEQ ID NO:17) and the mutant early H6 promoter (H6p*) (SEQ ID NO:18).
[0037]FIGS. 12A and 12B show the oligonucleotide primers used to amplify an H6p* 5'-FMDV fragment (SEQ ID NO:19-23) and the amino acids encoded by the nucleotides (SEQ ID NO:24 and 25).
[0038]FIGS. 13A and 13B show the construction of a pC6 H6p* FMDV P1+3C donor plasmid for generating ALVAC recombinants, with the insert at the unique C6 locus.
[0039]FIGS. 14A-14E show the nucleotide (SEQ ID NO:26) and amino acid sequences (SEQ ID NO:27) of the C6 H6p* FMDV gene cassette of the pC6 H6p* FMDV P1+3C donor plasmid.
[0040]FIGS. 15A and 15B show the oligonucleotide primers used to amplify the I3Lp 5'-FMDV fragment (SEQ ID NOS:28-33), and the amino acids encoded by the nucleotides (SEQ ID NO:34 and 35).
[0041]FIGS. 16A and 16B show the construction of a pC6 I3Lp FMDV P1+3C donor plasmid for generating ALVAC recombinants, with the insert at the unique C6 locus.
[0042]FIGS. 17A-17E show the nucleotide (SEQ ID NO:36) and amino acid sequences (SEQ ID NO:37) of the C6 I3Lp FMDV gene cassettes of the pC6 I3Lp FMDV P1+3C donor plasmid.
[0043]FIGS. 18A and 18B show the oligonucleotide primers used to amplify the 42Kp 5'-FMDV fragment (SEQ ID NO:38-43) and the amino acids encoded by the nucleotides (SEQ ID NO:44 and 45).
[0044]FIGS. 19A and 19B show the construction of a pC6 42Kp FMDV P1+3C donor plasmid for generating ALVAC recombinants, with the insert at the unique C6 locus.
[0045]FIGS. 20A-20E show the nucleotide (SEQ ID NO:46) and amino acid sequences (SEQ ID NO:47) of the C6 42Kp FMDV gene cassette of the pC6 42Kp FMDV P1+3C donor plasmid.
[0046]FIGS. 21A-21C show the oligonucleotide primers used to amplify and repair the 7.5Kp 5'-FMDV fragment (SEQ ID NO:48-54), and the amino acids encoded by the nucleotides (SEQ ID NO:55-57).
[0047]FIGS. 22A and 22B show the construction of a pC6 7.5K FMDV P1+3C donor plasmid for generating ALVAC recombinants, with the insert at the unique C6 locus.
[0048]FIGS. 23A-23E shows the nucleotide (SEQ ID NO:58) and amino acid sequences (SEQ ID NO:59) of the C6 7.5Kp FMDV gene cassette of the pC6 7.5Kp FMDV P1+3C donor plasmid.
[0049]FIGS. 24A-24E show the oligonucleotide primers used to amplify and repair the Pip 5'-FMDV fragment (SEQ ID NO:60-77), and the amino acids encoded by the nucleotides (SEQ ID NO:78-80).
[0050]FIGS. 25A and 25B show the construction of a pC6 Pip FMDV P1+3C donor plasmid for generating ALVAC recombinants, with the insert at the unique C6 locus.
[0051]FIGS. 26A-26E show the nucleotide (SEQ ID NO: 81) and amino acid sequences (SEQ ID NO:82) of the C6 Pip FMDV gene cassette of the pC6 Pip FMDV P1+3C donor plasmid.
[0052]FIG. 27 describes the oligonucleotide primers used to PCR amplify an H6p* 5'-FMDV fragment for insertion into a pF8 donor plasmid (SEQ ID NO:83-86).
[0053]FIGS. 28A and 28B illustrate the construction of a pF8 H6p* FMDV P1+3C donor plasmid for generating fowlpox recombinants.
[0054]FIGS. 29A-29F depict the nucleotide (SEQ ID NO:87) and amino acid (SEQ ID NO:88) sequences of the F8 H6p* FMDV P1+3C gene cassette of the pF8 H6p* FMDV P1+3C donor plasmid.
[0055]FIG. 30 shows the expression analysis of ALVAC recombinants containing the FMDV P1+3C gene cassette under the I3L or 42K promoters.
DETAILED DESCRIPTION OF THE INVENTION
[0056]In this disclosure, "comprises," "comprising," "containing" and "having" and the like can have the meaning ascribed to them in U.S. Patent law and can mean "includes," "including," and the like; "consisting essentially of" or "consists essentially" likewise has the meaning ascribed in U.S. Patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than that which is recited, but excludes prior art embodiments.
[0057]As used herein, the term "operably linked" means that the components described are in a relationship permitting them to function in their intended manner.
[0058]An "antigen" is a substance that is recognized by the immune system and induces an immune response. A similar term used in this context is "immunogen".
[0059]It is therefore an object of this invention to provide compositions and methods for treatment and prophylaxis of infection with FMDV. It is also an object to provide a means to treat or prevent foot and mouth disease.
[0060]In one aspect, the present invention relates to a modified recombinant avipox vector expressing at least one nucleic acid sequences encoding for one or more FMDV antigens. The viral vector according to the present invention is advantageously an avipox virus, such as fowlpox virus and canarypox virus and more particularly, ALVAC or TROVAC. The modified recombinant vector comprises a heterologous nucleic acid sequence, which encodes an antigenic protein, e.g., derived from FMDV ORFs that are encoded by the P1 (comprising VP1, VP2, VP3, VP4, and 2A), 2B, and/or 3C regions.
[0061]In another aspect, the present invention relates to a modified recombinant avipox virus that includes, in a non-essential region of the virus genome, at least one heterologous nucleic acid sequence that encodes one or more antigens from FMDV, such as gene products of the P1 gene (comprising VP1, VP2, VP3, VP4, 2A), 2B, and/or 3C.
[0062]In a still further aspect, the present invention relates to methods of eliciting an immune response to FMDV in a subject, comprising administering the recombinant avipox vector of the present invention. The present invention also relates to methods of eliciting an immune response to FMDV in a subject, comprising administering the recombinant avipox virus of the present invention. Advantageously, the avipox virus is selected from the group consisting of vCP2186, vCP2181, vCP2176, and vFP2215.
[0063]The virus used according to the present invention is advantageously a poxvirus, particularly an avipox virus, such as fowlpox virus or canarypox virus. TROVAC refers to an attenuated fowlpox that was a plaque-cloned isolate derived from the FP-1 vaccine strain of fowlpoxvirus that is licensed for vaccination of 1-day-old chicks. ALVAC is an attenuated canarypox virus-based vector that was a plaque-cloned derivative of the licensed canarypox vaccine, Kanapox (Tartaglia et al., 1992). ALVAC-based recombinant viruses expressing extrinsic immunogens have also been demonstrated efficacious as vaccine vectors (Tartaglia et al., 1993 a,b). This avipox vector is restricted to avian species for productive replication. On human cell cultures, canarypox virus replication is aborted early in the viral replication cycle prior to viral DNA synthesis. Nevertheless, when engineered to express extrinsic immunogens, authentic expression and processing is observed in vitro in mammalian cells and inoculation into numerous mammalian species induces antibody and cellular immune responses to the extrinsic immunogen and provides protection against challenge with the cognate pathogen (Taylor et al., 1992; Taylor et al., 1991).
[0064]ALVAC and TROVAC have also been recognized as unique among avipoxviruses in that the National Institutes of Health ("NIH"; U.S. Public Health Service), Recombinant DNA Advisory Committee, which issues guidelines for the physical containment of genetic material such as viruses and vectors, i.e., guidelines for safety procedures for the use of such viruses and vectors, which are based upon the pathogenicity of the particular virus or vector, granted a reduction in physical containment level: from BSL2 to BSL1. No other avipoxvirus has a BSL1 physical containment level. Even the Copenhagen strain of vaccinia virus--the common smallpox vaccine--has a higher physical containment level; namely, BSL2. Accordingly, the art has recognized that ALVAC and TROVAC have a lower pathogenicity than other avipox viruses.
[0065]Advantageously, the avipox virus vector is an ALVAC or a canarypox virus (Rentschler vaccine strain), which was attenuated through 200 or more serial passages on chick embryo fibroblasts, after which a master seed therefrom was subjected to four successive plaque purifications under agar, from which a clone was amplified through five additional passages. The avipox virus vector can also be a fowlpox virus, or an attenuated fowlpox virus such as TROVAC.
[0066]The invention further relates to the product of expression of the inventive recombinant avipox virus and uses therefor, such as to form antigenic, immunological or vaccine compositions for treatment, prevention, diagnosis or testing; and, to DNA from the recombinant avipox virus which are useful in constructing DNA probes and PCR primers.
[0067]In one aspect, the present invention relates to recombinant avipox viruses containing at least one nucleic acid sequence expressing one or more antigens from FMDV, advantageously in a non-essential region of the avipox virus genome. The avipox virus can be a fowlpox virus, especially an attenuated fowlpox virus such as TROVAC, or a canarypox virus, especially an attenuated canarypox virus, such as ALVAC.
[0068]According to the present invention, the recombinant avipox virus and avipox viral vectors express at least one nucleic acid sequence encoding one or more FMDV antigens. In particular, any or all genes or open reading frames (ORFs) encoding FMDV antigens can be isolated, characterized and inserted into ALVAC recombinants. The resulting recombinant avipox virus is used to infect an animal. Expression in the animal of FMDV antigens results in an immune response in the animal to FMDV. Thus, the recombinant avipox virus of the present invention may be used in an immunological composition or vaccine to provide a means to induce an immune response, which may, but need not be, protective. The molecular biology techniques used are described by Sambrook et al. (1989).
[0069]The invention also contemplates FMDV antigens that can be delivered as a naked DNA plasmid or vector, or DNA vaccine or immunological or immunogenic compositions comprising nucleic acid molecules encoding and expressing in vivo an FMDV antigen(s).
[0070]The FMDV antigen of interest can be obtained from FMDV or can be obtained from in vitro and/or in vivo recombinant expression of FMDV gene(s) or portions thereof. The FMDV antigen of interest can also be provided using synthetic FMDV sequences. The FMDV antigen of interest can be, but are not limited to: Lb, Lab, P1-2A (comprising VP1, VP2, VP3, VP4, and 2A); P2 (comprising 2B and 2C), and P3 (comprising 3A, 3B, VPg, 3C, and 3D), or portions thereof. In an advantageous embodiment, the FMDV antigens are P1 and 3C. In a particularly preferred embodiment, the FMDV antigens are P1-2A or P1-2A, 2B. Reference is made herein to U.S. patent application Ser. No. 10/327,481, relating to isolation of FMDV genome sequences, the contents of which are incorporated by reference.
[0071]Non-essential regions have been defined in the art (Johnson et al., (1993) Virology 196: 381-401) for vaccinia virus. These sites, also referred to herein as "insertion loci", are described in U.S. Pat. Nos. 6,340,462, and 5,756,103 for ALVAC, the contents of which are incorporated herein by reference, and include, but are not limited to, thymidine kinase (TK), hemagglutinin (HA), M2L, C6, and other loci. In one embodiment, where canarypox is used, the insertion locus is C6. In another embodiment, where fowlpox is used, the insertion locus is F8.
[0072]Insertion of nucleic acid sequences encoding FMDV antigens can be facilitated by homologous recombination, wherein the FMDV sequence of interest is flanked by sequences corresponding to avipox viral open reading frames immediately adjacent to the insertion locus (hereinafter referred to as "flanking sequences" or "insertion sequences"). Homologous recombination is facilitated by recognition of homologous flanking sequences, which promotes integration of the FMDV sequences into the insertion locus of interest. By way of example, insertion of FMDV sequences into the C6 locus requires the presence of the C6L and C6R ORFs on either side of the nucleic acid sequence encoding the FMDV antigen of interest in the viral vector. Thus, advantageously the insertion loci is C6 and the flanking sequences comprise C6L and C6R. Where the F8 insertion locus is used, the flanking sequences comprise F8L and F8R.
[0073]The recombinant viral vectors of the invention expressing FMDV antigens can be replicated or produced in cells or cell lines, or in vivo in a host or subject. One alternative embodiment consists of replicating the vector in cells permissive for replication of the vector.
[0074]It must be noted that avipox viruses can only productively replicate in or be passaged through avian species or avian cell lines such as, for example, chicken embryonic fibroblasts or QT35. The recombinant avipox viruses harvested from avian host cells, when inoculated into a non-avian vertebrate, such as a mammal, in a manner analogous to the inoculation of mammals by vaccinia virus, without productive replication of the avipox virus. Despite the failure of the avipox virus to productively replicate in such an inoculated non-avian vertebrate, sufficient expression of the virus occurs so that the inoculated animal responds immunologically to the antigenic determinants of the recombinant avipox virus and also to the antigenic determinants encoded in exogenous genes therein. Thus, in an advantageous embodiment, when avipox viruses or viral vectors are used, chicken embryonic fibroblasts or QT35 are preferred as the cells permissive for viral vector replication.
[0075]The recombinant viral vectors and recombinant viruses can contain promoters that are operably linked to the FMDV antigens of the present invention. The promoter is advantageously of poxyiral origin and advantageously early or early-late promoters, which may be, in particular, the promoter P11K of the vaccinia virus, I3L poxyiral promoter, 42K poxyiral promoter, H6 poxyiral promoter, Pi poxyiral promoter, P28K of the vaccinia virus, P160K ATI of the cowpox virus. In particular, the sequence driving the early transcription of an early-late promoter can be used instead of the full-length promoter (Moss, B. (1990) Ann. Rev. Biochem. 59: 661-688; Mars, M. et al, (1987) J. Mol. Biol. 198: 619-631; Davison, A. et al (1989) J. Mol. Biol. 210: 749-769; Vassef, A. (1987) Nucl. Acid. Res. 15: 1427-1443). The promoter is advantageously a weak promoter. The terms "strong promoter" and "weak promoter" are known in the art and are defined by the relative frequency of transcription initiation (times per minute) at the promoter.
[0076]The invention also provides for poxyiral promoters that are mutated. The present inventors have found that expression of certain FMDV antigens is not possible from strong poxyiral promoters. Without being bound by theory, it is believed that high levels of expression of potentially toxic FMDV antigens can preclude formation of stable poxyiral recombinants. Therefore, the present invention also comprehends the use of a mutated poxyiral promoter, such as, for example, a mutated H6 promoter, such that the expression levels of the FMDV antigens are decreased compared with expression levels of the FMDV antigens under a wild type promoter (Davison, A. et al (1989) J. Mol. Biol. 210: 749-769). The mutated H6 promoter of the instant invention can be considered a weak promoter.
[0077]The mutated H6 promoter taught herein contains a point mutation. The invention can also employ promoters other than H6, which contain point mutations that reduce their frequency of transcription initiation compared with the wild type promoter. In addition, other types of mutated promoters are suitable for use in the instant invention. For example, U.S. application Ser. No. 10/679,520, incorporated herein by reference, describes a truncated form of the H6 promoter (see also Davison, A. et al (1989) J. Mol. Biol. 210: 749-769; Taylor J. et al., Vaccine, 1988, 6, 504-508; Guo P. et al. J. Virol., 1989, 63, 4189-4198; Perkus M. et al., J. Virol., 1989, 63, 3829-3836).
[0078]The present invention also relates to a method of producing a recombinant avipox vector comprising FMDV antigens, comprising the steps of linearizing a donor plasmid with a restriction endonuclease, wherein the donor plasmid comprises restriction endonuclease cleavage sites or a multiple cloning site, and ligating at least one nucleic acid sequence comprising (i) a nucleic acid sequence encoding one or more FMDV antigen(s), (ii) a viral promoter sequence, and (iii) insertion sequences flanking (i) and (ii) that have complementary restriction endonuclease cleavage sites to the donor plasmid at FMDV antigens, thereby producing the recombinant avipox vector. Advantageously, the method further comprises the steps of introducing the vector into a cell permissive for replication of the vector, and isolating the vector from the cell.
[0079]By definition, a donor plasmid expression vector (or donor plasmid) includes a DNA transcription unit comprising a polynucleotide sequence containing the cDNA to be expressed and the elements necessary for its expression in vivo. The donor plasmid can also include a poxyiral early termination signal at the 3' terminus of the foreign gene (Moss, B. (1990) Ann. Rev. Biochem. 59: 661-688). The circular, super-coiled or uncoiled plasmid form is preferred. The linear form also comes under the scope of this invention.
[0080]Methods for making and/or using vectors (or recombinants) for expression and uses of expression products and products therefrom (such as antibodies) can be by or analogous to the methods disclosed in herein cited documents and documents referenced or cited in herein cited documents. See, for example, Sambrook et al. Molecular Cloning (1999). The invention also includes the use of the avipox vectors expressing FMDV antigens in the research setting. The recombinant avipox vectors and recombinant avipox viruses can be used to transfect or infect cells or cell lines of interest to study, for example, cellular responses to FMDV antigens, or signal transduction pathways mediated by FMDV antigens.
[0081]In the research setting, it is often advantageous to design recombinant vectors or viruses that comprise reporter genes that can be easily detected by laboratory assays and techniques. Reporter genes are well known in the art and can comprise resistance genes to antibiotics such as, but not limited to, ampicillin, neomycin, zeocin, kanamycin, bleomycin, hygromycin, chloramphenicol, among others. Reporter genes can also comprise green fluorescent protein, the lacZ gene (which encodes β-galactosidase), luciferase, and β-glucuronidase.
[0082]The invention also relates to a method of eliciting an immune response against foot-and-mouth disease in a subject comprising administering the recombinant avipox vectors or recombinant avipox viruses according to the present invention to the subject. The subject can be any animal which can become infected with FMDV, in particular, bovine, ovine, porcine or caprine species. Methods of administration and doses are defined herein.
[0083]The recombinant avipox vectors and viruses expressing FMDV antigens or an expression product thereof, immunological, antigenic or vaccine compositions or therapeutic compositions, can be administered via a parenteral route (intradermal, intramuscular or subcutaneous). Such an administration enables a systemic immune response, or humoral or cell-mediated responses.
[0084]As used herein, the terms "immunogenic composition" and "immunological composition" and "immunogenic or immunological composition" cover any composition that elicits an immune response against the targeted FMDV antigen; for instance, after administration of injection into the animal, elicits an immune response against the targeted FMDV antigen. The terms "vaccinal composition" and "vaccine" and "vaccine composition" covers any composition that induces a protective immune response against the FMDV antigen or which efficaciously protects against the antigen after administration or injection into the animal. The invention also comprehends recombinant avipox viral vectors administered as a plasmid DNA vector or vaccine.
[0085]More generally, the inventive recombinant avipox viral vectors and recombinant avipox viruses expressing FMDV antigens, antigenic, immunogenic, immunological or vaccine avipox virus-FMDV compositions or therapeutic compositions, can be prepared in accordance with standard techniques well known to those skilled in the pharmaceutical or veterinary arts. Such compositions can be administered in dosages and by techniques well known to those skilled in the medical or veterinary arts taking into consideration such factors as the age, sex, weight, species and condition of the particular patient, and the route of administration.
[0086]The compositions can be administered alone, or can be co-administered or sequentially administered with compositions, e.g., with "other" immunological, antigenic or vaccine or therapeutic compositions thereby providing multivalent or "cocktail" or combination compositions of the invention and methods of employing them. Again, the ingredients and manner (sequential or co-administration) of administration, as well as dosages can be determined taking into consideration such factors as the age, sex, weight, species and condition of the particular subject, and the route of administration. In this regard, reference is made to U.S. Pat. No. 5,843,456, incorporated herein by reference, and directed to rabies compositions and combination compositions and uses thereof.
[0087]Examples of compositions of the invention include liquid preparations for orifice, or mucosal, e.g., oral, nasal, anal, vaginal, peroral, intragastric, etc., administration such as suspensions, solutions, sprays, syrups or elixirs; and, preparations for parenteral, subcutaneous, intradermal, intramuscular or intravenous administration (e.g., injectable administration) such as sterile suspensions or emulsions. In such compositions, the recombinant avipox virus or recombinant avipox viral vectors may be in admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose or the like. The compositions can also be lyophilized. The compositions can contain auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, adjuvants, gelling or viscosity enhancing additives, preservatives, flavoring agents, colors, and the like, depending upon the route of administration and the preparation desired. Standard texts, such as "REMINGTON'S PHARMACEUTICAL SCIENCE", 17th edition, 1985, incorporated herein by reference, may be consulted to prepare suitable preparations, without undue experimentation.
[0088]Compositions in forms for various administration routes are envisioned by the invention. And again, the effective dosage and route of administration are determined by known factors, such as age, sex, weight, condition and nature of the animal, as well as LD50 and other screening procedures which are known and do not require undue experimentation. Dosages of each active agent can be as in herein cited documents (or documents referenced or cited in herein cited documents) and/or can range from one or a few to a few hundred or thousand micrograms, e.g., 1 μg to 1 mg, for an immunogenic, immunological or vaccine composition; and, 104 to 1010 TCID50 advantageously 106 to 103 TCID50 for an immunogenic, immunological or vaccine composition.
[0089]Recombinants or vectors can be administered in a suitable amount to obtain in vivo expression corresponding to the dosages described herein and/or in herein cited documents. For instance, suitable ranges for viral suspensions can be determined empirically. The viral vector or recombinant in the invention can be administered to an animal or infected or transfected into cells in an amount of about at least 103 pfu; more advantageously about 104 pfu to about 1010 pfu, e.g., about 105 pfu to about 109 pfu, for instance about 106 pfu to about 108 pfu, with doses generally ranging from about 106 to about 1010, advantageously about 108 pfu/dose, and advantageously about 107 pfu per dose of 2 ml. And, if more than one gene product is expressed by more than one recombinant, each recombinant can be administered in these amounts; or, each recombinant can be administered such that there is, in combination, a sum of recombinants comprising these amounts.
[0090]In vector or plasmid compositions employed in the invention, dosages can be as described in documents cited herein or as described herein or as in documents referenced or cited in herein cited documents. Advantageously, the dosage should be a sufficient amount of plasmid to elicit a response analogous to compositions wherein the antigen(s) of FMDV are directly present; or to have expression analogous to dosages in such compositions; or to have expression analogous to expression obtained in vivo by recombinant compositions. For instance, where DNA vaccines are administered, suitable quantities of each plasmid DNA in plasmid compositions can be 1 μg to 2 mg, advantageously 50 μg to 1 mg. Documents cited herein (or documents cited or referenced in herein cited documents) regarding DNA plasmid vectors may be consulted by the skilled artisan to ascertain other suitable dosages for DNA plasmid vector compositions of the invention, without undue experimentation.
[0091]However, the dosage of the composition(s), concentration of components therein and timing of administering the composition(s), which elicit a suitable immunological response, can be determined by methods such as by antibody titrations of sera, e.g., by ELISA and/or seroneutralization assay analysis. Such determinations do not require undue experimentation from the knowledge of the skilled artisan, this disclosure and the documents cited herein. And, the time for sequential administrations can be likewise ascertained with methods ascertainable from this disclosure, and the knowledge in the art, without undue experimentation.
[0092]The immunogenic or immunological compositions contemplated by the invention can also contain an adjuvant. Particularly suitable adjuvants for use in the practice of the present invention are (1) polymers of acrylic or methacrylic acid, maleic anhydride and alkenyl derivative polymers, (2) immunostimulating sequences (ISS), such as oligodeoxyribonucleotide sequences having one or more non-methylated CpG units (Klinman D. M. et al., Proc. Natl. Acad. Sci., USA, 1996, 93, 2879-2883; WO98/16247), (3) an oil in water emulsion, such as the SPT emulsion described on p 147 of "Vaccine Design, The Subunit and Adjuvant Approach" published by M. Powell, M. Newman, Plenum Press 1995, and the emulsion MF59 described on p 183 of the same work, (4) cationic lipids containing a quaternary ammonium salt, (5) cytokines, (6) aluminum hydroxide or aluminum phosphate or (7) other adjuvants discussed in any document cited and incorporated by reference into the instant application, or (8) any combinations or mixtures thereof. The DNA vaccines or immunogenic or immunological compositions encompassed by the invention can be formulated with a liposome, in the presence or absence of an adjuvant as described above.
[0093]Other suitable adjuvants include FMLP (N-formyl-methionyl-leucyl-phenylalanine; U.S. Pat. No. 6,017,537) and/or acrylic acid or methacrylic acid polymer and/or a copolymer of maleic anhydride and of alkenyl derivative. The acrylic acid or methacrylic acid polymers can be cross-linked, e.g., with polyalkenyl ethers of sugars or of polyalcohols. These compounds are known under the term "carbomer" (Pharmeuropa, Vol. 8, No. 2, June 1996). A person skilled in the art may also refer to U.S. Pat. No. 2,909,462 (incorporated by reference), which discusses such acrylic polymers cross-linked with a polyhydroxylated compound containing at least 3 hydroxyl groups: in one embodiment, a polyhydroxylated compound contains not more than 8 hydroxyl groups; in another embodiment, the hydrogen atoms of at least 3 hydroxyls are replaced with unsaturated aliphatic radicals containing at least 2 carbon atoms; in other embodiments, radicals contain from about 2 to about 4 carbon atoms, e.g., vinyls, allyls and other ethylenically unsaturated groups. The unsaturated radicals can themselves contain other substituents, such as methyl. The products sold under the name Carbopol® (Noveon Inc., Ohio, USA) are particularly suitable for use as an adjuvant. They are cross-linked with an allyl sucrose or with allylpentaerythritol, as to which, mention is made of the products Carbopol® 974P, 934P, and 971P.
[0094]As to the copolymers of maleic anhydride and of alkenyl derivative, mention is made of the EMA® products (Monsanto), which are copolymers of maleic anhydride and of ethylene, which may be linear or cross-linked, for example cross-linked with divinyl ether. Also, reference may be made to J. Fields et al., Nature 186:778-780, 1960 (incorporated by reference).
[0095]With regard to structure, the acrylic or methacrylic acid polymers and EMA are advantageously formed by basic units having the following formula:
##STR00001##
[0096]in which: [0097]R1 and R2, which can be the same or different, represent H or CH3 [0098]x=0 or 1, advantageously x=1 [0099]y=1 or 2, withx+y=2.
[0100]For EMA, x=0 and y=2 and for carbomers x=y=1.
[0101]These polymers are soluble in water or physiological salt solution (20 g/l NaCl) and the pH can be adjusted to 7.3 to 7.4, e.g., by soda (NaOH), to provide the adjuvant solution in which the expression vector(s) can be incorporated. The polymer concentration in the final vaccine composition can range between 0.01 and 1.5% w/v, advantageously 0.05 to 1% w/v and advantageously 0.1 to 0.4% w/v.
[0102]The cationic lipids containing a quaternary ammonium salt which are advantageously but not exclusively suitable for plasmids, are advantageously those having the following formula:
##STR00002##
in which R1 is a saturated or unsaturated straight-chain aliphatic radical having 12 to 18 carbon atoms, R2 is another aliphatic radical containing 2 or 3 carbon atoms and X is an amine or hydroxyl group.
[0103]Among these cationic lipids, preference is given to DMRIE (N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propane ammonium; WO96/34109), advantageously associated with a neutral lipid, advantageously DOPE (dioleoyl-phosphatidyl-ethanol amine; Behr J. P., 1994, Bioconjugate Chemistry, 5, 382-389), to form DMRIE-DOPE.
[0104]Advantageously, the plasmid mixture with the adjuvant is formed extemporaneously or contemporaneously with administration of the preparation or shortly before administration of the preparation; for instance, shortly before or prior to administration, the plasmid-adjuvant mixture is formed, advantageously so as to give enough time prior to administration for the mixture to form a complex, e.g. between about 10 and about 60 minutes prior to administration, such as approximately 30 minutes prior to administration.
[0105]When DOPE is present, the DMRIE:DOPE molar ratio is advantageously about 95:about 5 to about 5:about 95, more advantageously about 1:about 1, e.g., 1:1.
[0106]The DMRIE or DMRIE-DOPE adjuvant:plasmid weight ratio can be between about 50:about 1 and about 1:about 10, such as about 10:about 1 and about 1:about 5, and advantageously about 1:about 1 and about 1:about 2, e.g., 1:1 and 1:2.
[0107]A recombinant vaccine or immunogenic or immunological composition can also be formulated in the form of an oil-in-water emulsion. The oil-in-water emulsion can be based, for example, on light liquid paraffin oil (European Pharmacopea type); isoprenoid oil such as squalane, squalene, EICOSANE® or tetratetracontane; oil resulting from the oligomerization of alkene(s), e.g., isobutene or decene; esters of acids or of alcohols containing a linear alkyl group, such as plant oils, ethyl oleate, propylene glycol di(caprylate/caprate), glyceryl tri(caprylate/caprate) or propylene glycol dioleate; esters of branched fatty acids or alcohols, e.g., isostearic acid esters. The oil advantageously is used in combination with emulsifiers to form the emulsion. The emulsifiers can be nonionic surfactants, such as esters of sorbitan, mannide (e.g., anhydromannitol oleate), glycerol, polyglycerol, propylene glycol, and oleic, isostearic, ricinoleic, or hydroxystearic acid, which are optionally ethoxylated, and polyoxypropylene-polyoxyethylene copolymer blocks, such as the Pluronic® products, e.g., L121. The adjuvant can be a mixture of emulsifier(s), micelle-forming agent, and oil such as that which is available under the name Provax® (IDEC Pharmaceuticals, San Diego, Calif.).
[0108]The term "prime-boost" refers to the successive administrations of two different types of vaccine or immunogenic or immunological compositions having at least one antigen in common. The priming administration (priming) is the administration of a first vaccine or immunogenic or immunological composition type and may comprise one, two or more administrations. The boost administration is the administration of a second vaccine or immunogenic or immunological composition type and may comprise one, two or more administrations, and, for instance, may comprise or consist essentially of annual administrations.
[0109]Thus, the invention encompasses prime-boost immunization or vaccination method of an animal against at least one FMDV antigen comprising administering to the animal a priming DNA vaccine or immunological or immunogenic composition comprising nucleic acid molecule(s) encoding and expressing in vivo an antigen(s) from FMDV, and thereafter administering a boosting composition that comprises the FMDV antigen expressed by the DNA vaccine or immunogenic or immunological composition, or a recombinant or modified vector, e.g., virus, such as an avipox virus (such as ALVAC, canarypox, TROVAC, or fowlpox virus) that contains and expresses in an animal host cell a nucleotide sequence encoding the antigen of FMDV expressed by the DNA vaccine or immunogenic or immunological composition. The boosting vaccine or immunogenic or immunological composition can be the same as or different than the priming vaccine or immunogenic or immunological composition.
[0110]For instance, the boosting vaccine or immunogenic or immunological composition can be advantageously the FMDV antigen expressed by the DNA vaccine (or immunogenic or immunological composition) and/or a recombinant or modified avipox vector, e.g., virus, vaccine or immunogenic or immunological composition. A recombinant or modified vector is advantageously an in vivo expression vector, such as a modified or recombinant bacteria, yeast, virus, e.g. avipox virus, comprising nucleic acid molecule(s) encoding and expressing in vivo the antigen(s) from FMDV expressed by the DNA vaccine or immunogenic or immunological composition. The boost is advantageously performed with an inactivated vaccine or immunogenic or immunological composition, or with a vaccine or immunogenic or immunological composition comprising a recombinant live viral vector, such as a recombinant avipox virus, that comprises nucleic acid molecule(s) encoding and expressing in vivo the antigen(s) from the FMDV antigen expressed by the DNA vaccine or immunogenic or immunological composition. Thus, it is advantageous that the boost either comprises the antigen expressed by the DNA vaccine or immunogenic or immunological composition or expresses in vivo the same FMDV antigen expressed by the DNA vaccine or immunogenic or immunological composition. Advantageously, the boost comprises the recombinant avipox virus expressing FMDV antigens described herein.
[0111]Alternatively, the prime-boost immunization or vaccination method can comprise administering to the animal a priming vaccine comprising the recombinant avipox viruses of the present invention, and boosting thereafter with the DNA vaccine.
[0112]The DNA plasmid, or recombinant avipox vector expressing one or more nucleic acid sequences encoding at least one FMDV antigen, e.g., vector according to this disclosure, can be preserved and/or conserved and stored either in liquid form, at about 5° C., or in lyophilised or freeze-dried form, in the presence of a stabilizer. Freeze-drying can be according to well-known standard freeze-drying procedures. The pharmaceutically acceptable stabilizers may be SPGA (sucrose phosphate glutamate albumin; Bovarnik et al., J. Bacteriology 59:509, 1950), carbohydrates (e.g., sorbitol, mannitol, lactose, sucrose, glucose, dextran, trehalose), sodium glutamate (Tsvetkov T et al, Cryobiology 20(3): 318-23, 1983; Israeli E et al., Cryobiology 30(5): 519-23, 1993), proteins such as peptone, albumin or casein, protein containing agents such as skimmed milk (Mills C K et al, Cryobiology 25(2): 148-52, 1988; Wolff E et al., Cryobiology 27(5):569-75, 1990), and buffers (e.g., phosphate buffer, alkaline metal phosphate buffer). An adjuvant and/or a vehicle or excipient may be used to make soluble the freeze-dried preparations.
[0113]The invention will now be further described by way of the following non-limiting Examples, given by way of illustration.
EXAMPLES
Example 1
Construction of a pC5 H6p FMDV P1+3C Donor Plasmid for Introduction of FMDV Genes into the C5 Loci of ALVAC
[0114]Plasmid pAd5-A24 was used as the donor plasmid to generate the adenovirus Ad5A24 recombinant. It is a ˜39 kb plasmid containing the strain A24 P1 genes and the strain A12 3C protease. Several deletions of the FMDV genome were made for safety reasons and are indicated in FIG. 1.
[0115]Plasmid pAd5-A24 was digested with EcoRI and XbaT and the ˜3.4 kb fragment containing the FMDV genes was inserted in pUC8:2 (pUC8 with BglII and XbaI sites added to the multiple cloning site). The resulting 6 kb pUC FMDV plasmid (designated pHM-1119-1) was used as the source of the FMDV genes in all future constructs.
[0116]The H6 promoter (H6p) is an early/late promoter derived from the vaccinia H6 gene (Perkus, M. E. et al, (1989) J. Virol. 63: 3829-3836), which is designated as the H5 gene in the Copenhagen vaccinia strain. The H6p is a strong promoter that has been used extensively in avipox recombinants for foreign gene expression.
[0117]Plasmid pHM-1119-1 was used as the template for PCR amplification with primers 11277.SL and 11282.SL. These primers introduced the 3' end of the vaccinia H6 promoter, as well as translation and transcription stop signals, and XbaI or BamHI restriction sites for cloning. The primer sequences are shown in FIG. 2. The 3.4 kb PCR product was cloned into pCR2.1 to generate plasmid pHM-1151-4, pCR2.1 H6p FMDV.
[0118]Plasmid pCXL-148-2 is an ALVAC insertion plasmid for the C5 loci, which contains the vaccinia virus H6 promoter. The 3.4 kb NruI-XbaI fragment from pHM-1151-4 was inserted into pCXL-148-2, to generate pC5 H6p FMDV P1+3C (pHM-1175-1). The construction of pHM-1175-1 is illustrated in FIGS. 3A and 3B and the sequence of the C5 H6p FMDV gene cassette is shown in FIGS. 4A-4E.
[0119]Despite multiple attempts, no ALVAC recombinants were generated from pC5 H6p FMDV P1+3C, pHM-1175-1.
Example 2
Construction of a pF8 H6p FMDV P1+3C Donor Plasmid for Introduction of FMDV Genes into the F8 Locus of Fowlpox
[0120]Plasmid pSL-6427-2-1 (pF8 H6p) is a fowlpox insertion plasmid, which contains the vaccinia virus H6 promoter. The 3.4 kb NruI-BamHI fragment from pHM-1151-4 (pCR2.1 H6p FMDV; see Example 1) was inserted into pSL-6427-2-1, generating vector pHM-1180-11 (pF8 H6p FMDV P1+3C). The construction of pHM-1180-11 is illustrated in FIGS. 5A and 5B and the sequence of the F8 H6p FMDV gene cassette is shown in FIGS. 6A-6F.
[0121]Despite multiple attempts, no fowlpox recombinants could be generated from pF8 H6p FMDV P1+3C, pHM-1180-1.
Example 3
Construction of a Promoter-Less pC6 FMDV P1+3C Insertion Plasmid
[0122]The failure to generate avipox recombinants expressing FMDV genes could be due to the use of the strong vaccinia virus H6 promoter in the pC5 H6p FMDV P1+3C and pF8 H6p FMDV P1+3C plasmids described in Examples 1 and 2. In addition, the ALVAC donor plasmid results in the insertion of gene cassettes at the two C5 loci. For ALVAC, different viral promoters and the unique C6 insertion locus was used.
[0123]Plasmid pHM-1119-1 (pUC FMDV, see Example 1) was used as the template for PCR amplification of a 3'-fragment of FMDV, with primers 11280.SL and 11352.CXL. The ˜900 bp PCR fragment contains the 3'-end of FMDV from the XhoI site and introduces translational and transcriptional stops and a PstI cloning site. The primers are illustrated in FIG. 7. The PCR fragment was cloned into pCR2.1, generating plasmid pHM-1240-2, pCR2.1 3'-FMDV.
[0124]Plasmid pC6L is an ALVAC insertion plasmid for the unique ALVAC C6 site. The ˜2.6 kb EcoRI-Xho 5'-FMDV fragment from pHM-1119-1 was inserted into pC6L, generating plasmid pCXL-1008-1, pC6 5'-FMDV. The ˜900 bp XhoI-PstI fragment from pHM-1240-2 was inserted into pCXL-1008-1, generating pCXL-1013-2, pC6 FMDV. The construction of pC6 FMDV is illustrated in FIG. 8.
Example 4
Construction of a pC6 H6p FMDV P1+3C Donor Plasmid for Insertion of the FMDV Gene Cassette at the Unique C6 Locus of ALVAC
[0125]Plasmid pSL-6407-7 is a pC6 H6p insertion plasmid for the ALVAC C6 locus, which contains the vaccinia virus H6 promoter. The H6 promoter is in the opposite orientation to the C6 arms. The ˜2.6 kb NruI-Xho 5'-FMDV fragment from pHM-1151-4 (pCR2.1 FMDV, see Example 1) was inserted into pSL-6407-7, generating pC6 H6p 5'-FMDV, pCXL-1008-3. The ˜900 bp Xho-PstI 3'-FMDV fragment from pHM-1240-2 (pCR2.1 3'-FMDV, see Example 3) was inserted into pCXL-1008-3, generating pC6 H6p FMDV P1+3C, pCXL-1013-4. The construction of pC6 H6p FMDV P1+3C is illustrated in FIGS. 9A and 9B and the sequence of the C6 H6p FMDV gene cassette is shown in FIGS. 10A-10E.
[0126]Despite multiple attempts, ALVAC recombinants could not be generated using the pC6 H6p FMDV P1+3C donor plasmid, suggesting that insertion at a single site with a strong promoter was not feasible.
Example 5
Construction of a pC6 H6p* FMDV P1+3C Donor Plasmid for Insertion of the FMDV Gene Cassette at the Unique C6 Locus of ALVAC
[0127]Based upon studies with the vaccinia virus 7.5K early promoter (Davison, A. J. and Moss, B. (1989) J. Mol. Biol. 210: 749-769), a point mutation was introduced into the vaccinia virus H6 early promoter region, generating a mutant H6 promoter, H6p*. The wild-type early/late H6p and mutant early H6p* sequences are shown in FIG. 11.
[0128]Plasmid pHM-1119-1 (pUC FMDV, see Example 1) was used as the template to PCR amplify the H6p* 5'-FMDV fragment, with primers 11353.CXL and 11358.CXL. The ˜1.2 kb fragment contained the H6p* and the 5'-FMDV genes up to a unique NdeI site. The fragment was cloned into pCR2.1, generating plasmid pHM-1249-1-3. This clone was missing a nucleotide in VP4, so site-directed mutagenesis was performed with primers 11410.HM and 11411.HM to repair the PCR error. Clone pHM-1260-2, pCR2.1 H6p* 5'-FMDV, was confirmed by sequence analysis. FIG. 12A describes the PCR amplification primers and FIG. 12B describes the mutagenesis primers.
[0129]The ˜1.2 kb EcoR I-Nde I H6p* 5'-FMDV fragment from pHM-1260-2 was inserted into pCXL-1013-2 (pC6 FMDV P1+3C, see Example 3), generating plasmid pHM-1273-1, pC6 H6p* FMDV P1+3C. The construction of pC6 H6p* FMDV P1+3C is illustrated in FIGS. 13A and 13B and the sequence of the C6 H6p* FMDV gene cassette is shown in FIGS. 14A-14E.
[0130]To generate an ALVAC recombinant, primary chicken embryonic fibroblasts (CEF) were transfected with SapI-linearized pHM-1273-1 donor plasmid, in the presence of FuGENE-6® reagent (Roche). The transfected cells were subsequently infected with ALVAC as rescue virus at an MOI of 10 and after 24 hours, the transfected-infected cells were harvested, sonicated, and used for recombinant virus screening. Recombinant plaques were screened based on the plaque lift hybridization method using a 1.7 kb FMDV-specific probe labeled with horseradish peroxidase (HRP) according to the manufacturer's protocol (Amersham). ALVAC recombinants were generated and designated as vCP2176.
Example 6
Construction of a pC6 T3Lp FMDV P1+3C Donor Plasmid for Introduction of the FMDV Genes into the Unique C6 Locus of ALVAC
[0131]The early/intermediate I3L promoter (I3Lp) from vaccinia virus (Schmitt, J. F. and Stunnenberg, H. G. (1988) J. Virol. 62: 1889-1897) has been used previously in avipox recombinants.
[0132]Plasmid pCXL-1-4 is pC5 H6p EHV-1 gB (-TM)/42Kp EHV-1gD (-TM)/I3Lp EHV-1 gC (-TM), a donor plasmid used to introduce the EHV-1 gB, gC, and gD genes into ALVAC (described in U.S. Pat. No. 5,756,103). Each gene utilizes a different viral promoter, so pCXL-1-4 was used as the template to PCR amplify the I3L promoter. Primers 11407.CXL and 11423.CXL were used to amplify a 75 bp fragment containing the I3 L promoter and the 5'-end of the FMDV genes. The PCR primers are described in FIG. 15A.
[0133]A 648 bp PCR fragment, which contains a 20 bp overlap with the 75 bp I3Lp fragment, was amplified using primers 11425.CXL and 11407.CXL, with pHM-1119-1 (pUC FMDV, see Example 1) as template. This fragment contained the 5'-FMDV genes up to the unique KpnI site. The PCR primers are described in FIG. 15B.
[0134]The two PCR fragments were mixed at a 1:1 molar ratio and PCR amplified using primers 11423.CXL and 11407.CXL. The resultant 703 bp fragment was cloned into pCR2.1, generating pCXL-1068-1 (pCR2.1 I3Lp 5'-FMDV).
[0135]The ˜700 bp EcoRI-KpnI I3Lp 5'-FMDV fragment from pCXL-1068-1 was inserted into pHM1119-1, generating pCXL-1072-2 (pUC I3Lp FMDV P1+3C).
[0136]The ˜1.2 kb EcoRI-NdeI I3Lp 5'-FMDV fragment from pCXL-1072-2 was inserted into pCXL-1013-2 (pC6 FMDV P1+3C). The construction of pC6 I3Lp FMDV P1+3C is illustrated in FIGS. 16A and 16B and the sequence of the C6 I3Lp FMDV gene cassette is shown in FIGS. 17A-17E.
[0137]To generate an ALVAC recombinant, primary CEFs were transfected with 20 μg of SapI-linearized donor plasmid pCXL-1079-1 using FuGENE-6® reagent (Roche). The transfected cells were subsequently infected with ALVAC as rescue virus at an MOI of 10 and after 24 hours, the transfected-infected cells were harvested, sonicated, and used for recombinant virus screening. Recombinant plaques were screened based on the plaque lift hybridization method using a 1.7 kb FMDV-specific probe labeled with horseradish peroxidase (HRP) according to the manufacturer's protocol (Amersham). After four sequential rounds of plaque purification, the recombinants designated as vCP2181.4.1.1.1 and vCP2181.5.1.1.1 were generated and confirmed by hybridization as 100% positive for the FMDV insert and 100% negative for the C6 ORF.
[0138]Single plaques were selected from the 4th round of plaque purification and expanded to obtain P1 (1×T25 flask per sister), P2 (1×T75 flask per sister) and P3 (4×roller bottles per sister) amplified stocks of the vCP2181 recombinants. The infected cells from the roller bottles were harvested and concentrated to produce virus stock. The viral concentrate was re-confirmed by hybridization of plaque lifts with the FMDV- and C6-specific probes. Viral DNA was prepared and the correct insertion of the FMDV gene cassette at the ALVAC C6 locus was confirmed by Southern blot and sequence analyses. Immunoblot and immunoplaque assays were performed using specific antibodies as described in Example 7 (see FIG. 30).
Example 7
Construction of a pC6 42 kDp FMDV P1+3C Donor Plasmid for the Introduction of the FMDV Genes into the Unique C6 Locus of ALVAC
[0139]A 42K promoter (42Kp) derived from the AMV091 gene (vaccinia virus A23R homolog) of the insect poxvirus Amsacta moorei (Bawden, A. L. et al, (2000) Virology 274: 120-139) has been used previously in avipox recombinants (U.S. Pat. No. 5,756,103).
[0140]Plasmid pCXL-1-4 is pC5 H6p EHV-1 gB (-TM)/42Kp EHV-1 gD (-TM)/I3Lp EHV-1 gC (-TM), a donor plasmid used to introduce the EHV-1 gB, gC, and gD genes into ALVAC (see Example 6). Each gene uses a different viral promoter, so pCXL-1-4 was used as the template to PCR amplify the 42K promoter. Primers 11426.CXL and 11427.CXL were used to amplify a 48 bp fragment containing the 42K promoter and the 5'-end of the FMDV genes. The PCR primers are described in FIG. 18A.
[0141]A 647 bp PCR fragment, which contains a 20-bp overlap with the 48 bp 42Kp fragment, was amplified using primers 11428.CXL and 11407.CXL, with pHM-1119-1 (pUC FMDV, see Example 1) as a template. This fragment contains the 5'-FMDV genes up to the unique KpnI site. The PCR primers are described in FIG. 18B.
[0142]The two PCR fragments were mixed at a 1:1 molar ratio and PCR amplified using primers 11426.CXL and 11407.CXL. The resultant 676 bp fragment was cloned into pCR2.1, generating pCXL-1080-2-2 (pCR2.1 42Kp 5'-FMDV).
[0143]The 676 bp EcoRI-KpnI 42Kp 5'-FMDV fragment from pCXL-1080-2-2 was inserted into pHM-1119-1, generating pCXL-1089-1 (pUC 42Kp FMDV P1+3C).
[0144]The ˜1.2 kb EcoRI-NdeI 42Kp 5'-FMDV fragment from pCXL-1089-1 was inserted into pCXL-1013-2 (pC6 FMDV P1+3C, see Example 3), generating pCXL-1095-1 (pC6 42Kp FMDV P1+3C). The construction of pC6 42Kp FMDV P1+3C is illustrated in FIGS. 19A and 19B and the sequence of the C6 42Kp FMDV gene cassette is shown in FIGS. 20A-20E.
[0145]To generate an ALVAC recombinant, primary CEFs were transfected with 20 μg of SapI-linearized donor plasmid pCXL-1095-1, using FuGENE-6® reagent (Roche). The transfected cells were subsequently infected with ALVAC as rescue virus at an MOI of 10 and after 29 hours, the transfected-infected cells were harvested, sonicated, and used for recombinant virus screening. Recombinant plaques were screened based on the plaque lift hybridization method using the 1.7 kb FMDV-specific probe labeled with horseradish peroxidase (HRP) according to the manufacturer's protocol (Amersham). After four sequential rounds of plaque purification, the recombinant designated as vCP2186.6.2.1.1 was generated and confirmed by hybridization as 100% positive for the FMDV insert and 100% negative for the C6 ORF.
Single plaques were selected from the 4th round of plaque purification, and expanded to obtain P1 (1×T25 flask), P2 (1×T75 flask) and P3 (8×roller bottles) amplified stocks. The infected cells from the roller bottles were harvested and concentrated to produce virus stock. The virl concentrate was characterized by performing hybridization of plaque lifts with the FMDV- and C6-specific probes to confirm 100% genetic purity. Viral DNA was extracted and Southern blotting and sequence analyses confirmed the correct insertion of the FMDV gene cassette.
[0146]For expression analysis, CEFs were infected at an MOI of 10 with vCP2181 (ALVAC C6 I3Lp FMDV P1+3C; see Example 6) or vCP2186 (ALVAC C6 42Kp FMDV P1+3C) and grown at 37° C., in the presence of 5% CO2, for 24 hours. The supernatant was harvested and clarified and the cell monolayer was resuspended in PBS, and then pelleted. The pellets were resuspended in water, and then SDS PAGE sample buffer was added to the supernatants. The protein samples were separated on a 10% SDS PAGE gel, then electrotransferred to a nylon membrane. The membrane was blocked, and then probed with rabbit anti-FMDV VP1, VP2, and VP3 antisera. Secondary antibody and colorimetric analysis revealed that both recombinants expressed specific proteins of sizes consistent with VP0, VP1 and VP3 in both the pellets and supernatants. These data are illustrated in FIG. 30.
Example 8
Construction of a pC6 7.5Kp FMDV P1+3C Donor Plasmid for the Introduction of the FMDV Genes into the Unique C6 Locus of ALVAC
[0147]The early 7.5K promoter (7.5Kp) of vaccinia virus (Davison, A. J. and Moss, B. (1989) J. Mol. Biol. 210: 749-769) has been used previously in avipox recombinants.
[0148]Plasmid pHM-1119-1 (pUC FMDV, see Example 1) was used as the template for PCr amplification of the 7.5K promoter and FMDV genes, up to the unique NdeI site. Primers 11357.CXL and 11358.CXL were used to amplify a 1214 bp 7.5Kp 5'-FMDV fragment, which was cloned into pCR2.1, generating pHM-1249-5-3. The PCR amplification primers are describe in FIG. 21A.
[0149]Sequence analysis revealed that three base pair deletions in pHM-1249-5-3. Oligonucleotide primers 11429.HM and 11430.HM were designed to re-introduce the missing ucleotides by site-directed mutagenesis. The mutagenesis primers are described I FIG. 21B. The resultant clones contained 2 of the 3 re-introduced nucleotides, so clone pHM-1267-4 was subjected to a further round of site-directed mutagenesis with primers 11445.HM and 11446.HM. The mutagenesis primers are described in FIG. 21C. Clone pHM-1299-2 (pCR2.1 7.5Kp 5'-FMDV) was confirmed to be correct by sequence analysis.
[0150]The ˜1.2 kb EcoRI-NdeI fragment from pHM-1299-2 was inserted into pCXL-1013-2 (pC6 FMDV, see Example 3), generating plasmid pHM-1310-4 (pC6 7.5Kp FMDV P1+3C). The construction of pHM-1310-4 is illustrated in FIGS. 22A and 22B and the sequence of the C6 7.5Kp FMDV gene cassette is shown in FIGS. 23A-23E.
[0151]ALVAC recombinant vCP2189 was obtained after two rounds of screening, but could not be purified/amplified and was lost, suggesting that it was unstable and/or toxic.
Example 9
Construction of a pC6 Pip FMDV P1+3C Donor Plasmid for the Insertion of the FMDV Genes into the Unique C6 Locus of ALVAC
[0152]The early Pi promoter (Pip) from vaccinia virus (Wachsman, M. et al, (1987) J. Infect. Dis. 155: 1188-1197) has been used previously in avipox recombinants. It is 81 nucleotides in length and is thought to be a relatively weak promoter.
[0153]Plasmid pHM-1119-1 (pUC FMDV, see Example 1) was used as a template to PCR-amplify the Pip 5'-FMDV fragment, with primers 11356.CXL and 11358.CXL (FIG. 24A). The amplified fragment was cloned into pCR2.1 and several clones were screened by sequence analysis. The clone with the fewest PCR errors (pHM-1249-4-4, pCR2.1 Pip* 5'-FMDV) was missing 28 nucleotides randomly throughout the Pi promoter region, including the EcoRI cloning site.
[0154]Oligonucleotides 11395.CXL and 11399.CXL (FIG. 24B) were used to assemble the correct Pi promoter. The Pip was PCR amplified with primers 11400.CXL and 11401.CXL (FIG. 24C) and cloned into pCR2.1 to generate pHM-1263-1 (pCR2.1 Pip). Plasmid pHM-1263-1 was used as a template to PCR-amplify a 97 bp Pip 5'-FMDV fragment, using primers 11402.CXL and 1140.CXL (FIG. 24D). This fragment contains the EcoRI cloning site, the full-length Pip and 10 bp of FMDV.
[0155]Using pHM-1249-4-4 as template, a 648 bp fragment was PCR-amplified using primers 11406.CXL and 11407.CXL (FIG. 24E). This fragment contains 10 bp of the 3' end of Pip and the 5'-FMDV genes up to a unique KpnI site.
[0156]Equimolar amounts of the 97 bp Pip 5'-FMDV and 648 bp 3'-Pip 5'-FMDV PCR fragments were mixed and amplified using primers 11402.CXL and 11407.CXL. The resulting 745 bp Pip 5'-FMDV (EcoRI-KpnI) fragment was cloned into pCR2.1 to generate pHM-1268-1 (pCR2.1 Pip 5'-FMDV, EcoRI-KpnI). The EcoRI-KpnI fragment from pHM-1268-1 was inserted into pHM-1119-1, generating pHM-1277-6 (pUC Pip FMDV).
[0157]The 1252 bp EcoRI-NdeI Pip 5'-FMDV fragment from pHM-1277-6 was inserted into plasmid pCXL-1013-2 (pC6 FMDV P1+3C, see Example 3), generating plasmid pHM-1284-25 (pC6 Pip FMDV P1+3C). The construction of pC6 Pip FMDV P1+3C is illustrated in FIG. 25 and the sequence of the C6 Pip FMDV gene cassette is shown in FIGS. 26A-26E.
[0158]ALVAC recombinant vCP2184 was obtained after two rounds of purification, but was lost at the third round of screening/amplification, suggesting that it was toxic and/or unstable.
Example 10
Construction of a pF8 H6p* FMDV P1+3C Donor Plasmid for Insertion of the FMDV Gene Cassette at the Unique F8 Locus of Fowlpox
[0159]Plasmid pHM-1260-2 (pCR2.1 H6p* 5'-FMDV (Nde); see Example 5) was used as the template to PCR amplify an H6p* 5'-FMDV fragment, with primers 11506.HM and 11279.5L. Primer 11506.HM was designed to introduce a Hind III site in front of H6p* and primer 11279.5L was designed to amplify the FMDV genes up to the unique KpnI site in the VP2 gene. The ˜700 bp fragment was cloned into pCR2.1, generating pHM-1341-7 (pCR2.1 H6p* 5'-FMDV KpnI), which was confirmed as correct by sequence analysis. FIG. 27 describes the PCR primers.
[0160]Plasmid pHM-1180-11 is pF8 H6p FMDV P1+3C, containing the wild-type H6 promoter (see Example 2 and FIGS. 5A and 5B). Plasmid pSL-6427-1-1 (pF8 MCS) is a promoter-less plasmid used for insertion into the fowlpox F8 site. The 0.7 kb HindIII-KpnI H6p* 5'-FMDV fragment from pHM-1341-7 and the 2.7 kb KpnI-BamH I 3'-FMDV fragment from pHM-1180-11 were ligated into plasmid pSL-6427-1-1 that had been digested with HindIII and BamHI, generating pHM-1354-1 (pF8 H6p* FMDV P1+3C). The construction ofpHM-1354-1 is illustrated in FIGS. 28A and 28B and the sequence of the F8 H6p* FMDV P1+3C gene cassette is shown in FIGS. 29A-29F.
[0161]To generate a fowlpox recombinant, primary CEFs were transfected with NotI-linearized pHM-1354-1, in the presence of Fugene-6® reagent (Roche). The transfected cells were subsequently infected with fowlpox as rescue virus at MOI of 10 and after 51 hours, the transfected-infected cells were harvested, sonicated and used for recombinant virus screening. Recombinant plaques were screened based on the plaque lift hybridization method using a 1.7 kb FMDV-specific probe labelled with horseradish peroxidase (HRP) according to the manufacturer's protocol (Amersham Cat# RPN3001). After five sequential rounds of plaque purification, a fowlpox recombinant designated as vFP2215.1.3.1.1.1 was generated and confirmed by hybridization as 100% positive for the FMDV insert and 100% negative for the F8 ORF.
[0162]Single plaques were selected from the 5th round of plaque purification, and expanded to obtain P1 (1×T25 flask), P2 (2×T75 flask) and P3 (10×roller bottles) stocks to amplify vFP2215. The infected cells from the roller bottles was harvested and concentrated to produce virus stock. The viral concentrate was re-confirmed by hybridization of plaque lifts with the FMDV- and F8-specific probes. Viral DNA was prepared and the correct insertion of the FMDV gene cassette at the fowlpox F8 locus was confirmed by Southern blot and sequence analyses.
Example 11
Preparation and Purification of Plasmids
[0163]For the preparation of the plasmids intended for the vaccination of animals, any technique may be used which makes it possible to obtain a suspension of purified plasmids predominantly in a supercoiled form. These techniques are well known to persons skilled in the art. There may be mentioned in particular the alkaline lysis technique followed by two successive ultracentrifugations on a caesium chloride gradient in the presence of ethidium bromide as described in J. Sambrook et al. (Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1989). Reference may also be made to Patent Applications PCT WO 95/21250 and PCT WO 96/02658, which describe methods for producing, on an industrial scale, plasmids which can be used for vaccination. For the purposes of the manufacture of vaccines (see Example 11), the purified plasmids are resuspended so as to obtain solutions at a high concentration (>2 mg/ml), which are compatible with storage. To do this, the plasmids are resuspended either in ultrapure water or in TE buffer (10 mM Tris-HC 1; 1 mM EDTA, pH 8.0).
Example 12
Manufacture of the Associated Vaccine
[0164]The various plasmids necessary for the manufacture of an associated vaccine are mixed starting with their concentrated solutions (Example 10). The mixtures are prepared such that the final concentration of each plasmid corresponds to the effective dose of each plasmid. The solutions, which can be used to adjust the final concentration of the vaccine may be either a 0.9M NaCl solution, or PBS buffer.
[0165]Specific formulations such as liposomes or cationic lipids may also be used for the manufacture of the vaccines.
Example 13
Vaccination of Animals
[0166]The animals are vaccinated with doses of 100 pg, 250 μg or 500 μg per plasmid. The injections are performed with a needle by the intramuscular route either at the level of the gluteus muscle, or at the level of the neck muscles. The vaccinal doses are administered in volumes of between 1 and 5 ml.
[0167]Having thus described in detail preferred embodiments of the present invention, it is to be understood that the invention defined by the appended claims is not to be limited to particular details set forth in the above description as many apparent variations thereof are possible without departing from the spirit or scope of the present invention.
Sequence CWU
1
88154DNAArtificial SeqenceDescription of Artificial Sequence Synthetic
primer 1tatcgcgata tccgttaagt ttgtatcgta atgggagctg ggcaatccag ccca
54246DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 2gttgaccctg aaccacaaca cgagtagtaa tttttctaga ggatcc
46346DNAArtificial SequenceDescription of Artificial
Sequence Synthetic primer 3ggatcctcta gaaaaattac tactcgtgtt
gtggttcagg gtcaac 4648PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 4Met
Gly Ala Gly Gln Ser Ser Pro1 558PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 5Val
Asp Pro Glu Pro Gln His Glu1 565594DNAFoot-and-mouth
disease virusCDS(1758)..(5135) 6tgaatgttaa atgttatact ttggatgaag
ctataaatat gcattggaaa aataatccat 60ttaaagaaag gattcaaata ctacaaaacc
taagcgataa tatgttaact aagcttattc 120ttacgacgc tttaaatata cacaaataaa
cataattttt gtataaccta acaaataact 180aaaacataaa aataataaaa ggaaatgtaa
tatcgtaatt attttactca ggaatggggt 240taaatattta tatcacgtgt atatctatac
tgttatcgta tactctttac aattactatt 300acgaatatgc aagagataat aagattacgt
atttaagaga atcttgtcat gataattggg 360tacgacatag tgataaatgc tatttcgcat
cgttacataa agtcagttgg aaagatggat 420ttgacagatg taacttaata ggtgcaaaaa
tgttaaataa cagcattcta tcggaagata 480ggataccagt tatattatac aaaaatcact
ggttggataa aacagattct gcaatattcg 540taaaagatga agattactgc gaatttgtaa
actatgacaa taaaaagcca tttatctcaa 600cgacatcgtg taattcttcc atgttttatg
tatgtgtttc agatattatg agattactat 660aaactttttg tatacttata ttccgtaaac
tatattaatc atgaagaaaa tgaaaaagta 720tagaagctgt tcacgagcgg ttgttgaaaa
caacaaaatt atacattcaa gatggcttac 780atatacgtct gtgaggctat catggataat
gacaatgcat ctctaaatag gtttttggac 840aatggattcg accctaacac ggaatatggt
actctacaat ctcctcttga aatggctgta 900atgttcaaga ataccgaggc tataaaaatc
ttgatgaggt atggagctaa acctgtagtt 960actgaatgca caacttcttg tctgcatgat
gcggtgttga gagacgacta caaaatagtg 1020aaagatctgt tgaagaataa ctatgtaaac
aatgttcttt acagcggagg ctttactcct 1080ttgtgtttgg cagcttacct taacaaagtt
aatttggtta aacttctatt ggctcattcg 1140gcggatgtag atatttcaaa cacggatcgg
ttaactcctc tacatatagc cgtatcaaat 1200aaaaatttaa caatggttaa acttctattg
aacaaaggtg ctgatactga cttgctggat 1260aacatgggac gtactccttt aatgatcgct
gtacaatctg gaaatattga aatatgtagc 1320acactactta aaaaaaataa aatgtccaga
actgggaaaa attgatcttg ccagctgtaa 1380ttcatggtag aaaagaagtg ctcaggctac
ttttcaacaa aggagcagat gtaaactaca 1440tctttgaaag aaatggaaaa tcatatactg
ttttggaatt gattaaagaa agttactctg 1500agacacaaaa gaggtagctg aagtggtact
ctcaaaggta cgtgactaat tagctataaa 1560aaggatccgg gttaattaat tagtcatcag
gcagggcgag aacgagacta tctgctcgtt 1620aattaattag agcttcttta ttctatactt
aaaaagtgaa aataaataca aaggttcttg 1680agggttgtgt taaattgaaa gcgagaaata
atcataaatt atttcattat cgcgatatcc 1740gttaagtttg tatcgta atg gga gct ggg
caa tcc agc cca gca acc ggc 1790 Met Gly Ala Gly
Gln Ser Ser Pro Ala Thr Gly 1 5
10tcg cag aac cag tct ggc aac act ggc agc ata atc aac aac tac tac
1838Ser Gln Asn Gln Ser Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr
15 20 25atg caa cag tac cag
aac tcc atg gac aca cag ttg gga gac aat gcc 1886Met Gln Gln Tyr Gln
Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala 30 35
40atc agt gga ggc tcc aac gag ggc tcc acg gac aca act
tca aca cac 1934Ile Ser Gly Gly Ser Asn Glu Gly Ser Thr Asp Thr Thr
Ser Thr His 45 50 55 aca acc aac act
caa aac aat gac tgg ttc tcg aag ctc gcc agt tca 1982Thr Thr Asn Thr
Gln Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser60 65
70 75gct ttt acc ggt ctg ttc ggt gca ctg
ctc gcc gac aag aag aca gag 2030Ala Phe Thr Gly Leu Phe Gly Ala Leu
Leu Ala Asp Lys Lys Thr Glu 80 85
90gaa acg aca ctt ctt gag gac cgc atc ctc acc acc cgc aac ggg
cac 2078Glu Thr Thr Leu Leu Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly
His 95 100 105acc acc tcg acg
acc caa tcg agt gtg ggt gtc aca cac ggg tac tcc 2126Thr Thr Ser Thr
Thr Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser 110
115 120aca gag gag gac cac gtt gct ggg ccc aac aca tcg
ggc ctg gag acg 2174Thr Glu Glu Asp His Val Ala Gly Pro Asn Thr Ser
Gly Leu Glu Thr 125 130 135cga gtg gtg
cag gca gag aga ttc tac aaa aag tac ttg ttt gac tgg 2222Arg Val Val
Gln Ala Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp140
145 150 155aca acg gac aag gca ttt gga
cac ctg gaa aag ctg gag ctc ccg tcc 2270Thr Thr Asp Lys Ala Phe Gly
His Leu Glu Lys Leu Glu Leu Pro Ser 160
165 170gac cac cac ggt gtc ttt gga cac ttg gtg gac tcg
tac gcc tat atg 2318Asp His His Gly Val Phe Gly His Leu Val Asp Ser
Tyr Ala Tyr Met 175 180 185aga
aat ggc tgg gat gtt gag gtg tcc gct gtt ggc aac cag ttc aac 2366Arg
Asn Gly Trp Asp Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn 190
195 200ggc ggg tgc ctc ctg gtg gcc atg gta
cct gaa tgg aag gaa ttt gac 2414Gly Gly Cys Leu Leu Val Ala Met Val
Pro Glu Trp Lys Glu Phe Asp 205 210
215aca cgg gag aaa tac caa ctc acc ctt ttc ccg cac cag ttt att agc
2462Thr Arg Glu Lys Tyr Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser220
225 230 235ccc aga act aac
atg act gcc cac atc acg gtc ccc tac ctt ggt gtg 2510Pro Arg Thr Asn
Met Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val 240
245 250aac agg tat gat cag tac aag aag cat aag
ccc tgg aca ttg gtt gtc 2558Asn Arg Tyr Asp Gln Tyr Lys Lys His Lys
Pro Trp Thr Leu Val Val 255 260
265atg gtc gtg tcg cca ctt acg gtc aac aac act agt gcg gca caa atc
2606Met Val Val Ser Pro Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile
270 275 280aag gtc tac gcc aac ata gct
ccg acc tat gtt cac gtg gcc ggt gaa 2654Lys Val Tyr Ala Asn Ile Ala
Pro Thr Tyr Val His Val Ala Gly Glu 285 290
295ctc ccc tcg aaa gag ggg att ttc ccg gtt gca tgt gcg gac ggt tac
2702Leu Pro Ser Lys Glu Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr300
305 310 315gga gga ttg gtg
acg aca gac ccg aag aca gct gac cct gct tat ggc 2750Gly Gly Leu Val
Thr Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly 320
325 330aag gtg tac aac ccg cct agg act aac tac
cct ggg cgc ttc acc aac 2798Lys Val Tyr Asn Pro Pro Arg Thr Asn Tyr
Pro Gly Arg Phe Thr Asn 335 340
345ctg ttg gac gtg gcc gaa gcg tgt ccc act ttc ctc tgc ttt gac gac
2846Leu Leu Asp Val Ala Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp
350 355 360ggg aaa ccg tac gtc acc acg
cgg acg gat gac acc cga ctt ttg gcc 2894Gly Lys Pro Tyr Val Thr Thr
Arg Thr Asp Asp Thr Arg Leu Leu Ala 365 370
375aag ttt gac ctt tcc ctt gcc gca aaa cat atg tcc aac aca tac ctg
2942Lys Phe Asp Leu Ser Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu380
385 390 395tca ggg att gct
cag tac tac aca cag tac tct ggc acc atc aat ttg 2990Ser Gly Ile Ala
Gln Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu 400
405 410cat ttc atg ttt aca ggt tcc act gat tca
aag gcc cga tac atg gtg 3038His Phe Met Phe Thr Gly Ser Thr Asp Ser
Lys Ala Arg Tyr Met Val 415 420
425gcc tac atc cca cct ggg gtg gag aca cca ccg gac aca cct gaa agg
3086Ala Tyr Ile Pro Pro Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg
430 435 440gct gcc cac tgc att cac gct
gaa tgg gac act gga cta aac tcc aaa 3134Ala Ala His Cys Ile His Ala
Glu Trp Asp Thr Gly Leu Asn Ser Lys 445 450
455ttc act ttc tca atc ccg tac gta tcc gcc gcg gat tac gcg tac aca
3182Phe Thr Phe Ser Ile Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr Thr460
465 470 475gcg tct gac acg
gca gaa aca atc aac gta cag gga tgg gtc tgc atc 3230Ala Ser Asp Thr
Ala Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile 480
485 490tac caa att aca cac ggg aag gct gaa aat
gac acc ttg gtc gtg tcg 3278Tyr Gln Ile Thr His Gly Lys Ala Glu Asn
Asp Thr Leu Val Val Ser 495 500
505gtt agc gcc ggc aaa gac ttt gag ttg cgc ctc ccg att gac ccc cgc
3326Val Ser Ala Gly Lys Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg
510 515 520cag cag acc acc gct acc ggg
gaa tca gca gac ccg gtc acc acc acc 3374Gln Gln Thr Thr Ala Thr Gly
Glu Ser Ala Asp Pro Val Thr Thr Thr 525 530
535gtg gag aac tac ggc ggt gag aca caa atc cag aga cgt cac cac acg
3422Val Glu Asn Tyr Gly Gly Glu Thr Gln Ile Gln Arg Arg His His Thr540
545 550 555gac att ggt ttc
atc atg gac aga ttt gtg aag atc caa agc ttg agc 3470Asp Ile Gly Phe
Ile Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser 560
565 570cca aca cat gtc att gac ctc atg cag gct
cac caa cac ggt ctg gtg 3518Pro Thr His Val Ile Asp Leu Met Gln Ala
His Gln His Gly Leu Val 575 580
585ggt gcc ttg ctg cgt gca gcc acg tac tac ttt tct gac ctg gaa att
3566Gly Ala Leu Leu Arg Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile
590 595 600gtt gta cgg cac gaa ggc aat
ctg acc tgg gtg ccc aac ggc gcc cct 3614Val Val Arg His Glu Gly Asn
Leu Thr Trp Val Pro Asn Gly Ala Pro 605 610
615gaa tca gcc ctg ttg aac acc agc aac ccc act gcc tac aac aag gca
3662Glu Ser Ala Leu Leu Asn Thr Ser Asn Pro Thr Ala Tyr Asn Lys Ala620
625 630 635cca ttc acg aga
ctc gct ctc ccc tac act gcg ccg cac cgt gtg ctg 3710Pro Phe Thr Arg
Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu 640
645 650gca aca gtg tac aac ggg acg agt aag tat
gct gtg ggt ggt tca ggc 3758Ala Thr Val Tyr Asn Gly Thr Ser Lys Tyr
Ala Val Gly Gly Ser Gly 655 660
665aga aga ggc gac atg ggg tct ctc gcg gcg cga gtc gtg aaa cag ctt
3806Arg Arg Gly Asp Met Gly Ser Leu Ala Ala Arg Val Val Lys Gln Leu
670 675 680cct gct tca ttt aac tac ggt
gca atc aag gcc gac gcc atc cac gaa 3854Pro Ala Ser Phe Asn Tyr Gly
Ala Ile Lys Ala Asp Ala Ile His Glu 685 690
695ctt ctc gtg cgc atg aaa cgg gcc gag ctc tac tgc ccc aga ccg ctg
3902Leu Leu Val Arg Met Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu700
705 710 715ttg gca ata gag
gtg tct tcg caa gac agg cac aag caa aag atc att 3950Leu Ala Ile Glu
Val Ser Ser Gln Asp Arg His Lys Gln Lys Ile Ile 720
725 730gca cca gca aag cag ctt ctg aat ttt gac
ctg ctc aag ttg gcc gga 3998Ala Pro Ala Lys Gln Leu Leu Asn Phe Asp
Leu Leu Lys Leu Ala Gly 735 740
745gac gtt gag tcc aac ccc ggg cca ttc ttc ttt gct gac gtt agg tca
4046Asp Val Glu Ser Asn Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser
750 755 760aac ttt tca aag ttg gta gac
aca atc aac cag atg cag gag gac atg 4094Asn Phe Ser Lys Leu Val Asp
Thr Ile Asn Gln Met Gln Glu Asp Met 765 770
775tcc aca aaa cac ggg ccc gac ttc aac cgg ttg gtg tcc gca ttt gag
4142Ser Thr Lys His Gly Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu780
785 790 795gaa ttg gcc act
gga gtt aaa gct atc agg acc ggt ctc gac gag gcc 4190Glu Leu Ala Thr
Gly Val Lys Ala Ile Arg Thr Gly Leu Asp Glu Ala 800
805 810aaa ccc tgg tac aag ctt atc aaa ctc cta
agc cgc ctg tcg tgc atg 4238Lys Pro Trp Tyr Lys Leu Ile Lys Leu Leu
Ser Arg Leu Ser Cys Met 815 820
825gcc gct gtg gca gca cgg tcc aag gac cca gtc ctt gtg gcc atc atg
4286Ala Ala Val Ala Ala Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met
830 835 840ctg gcc gac acc ggt ctc gag
cgt cag aga cct ctg aaa gtg aga gct 4334Leu Ala Asp Thr Gly Leu Glu
Arg Gln Arg Pro Leu Lys Val Arg Ala 845 850
855aag ctc cca cag cag gaa gga cct tac gct ggc ccg ttg gag aga cag
4382Lys Leu Pro Gln Gln Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln860
865 870 875aaa ccg ctg aaa
gtg aaa gca aaa gcc ccg gtc gtc aag gaa gga cct 4430Lys Pro Leu Lys
Val Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro 880
885 890tac gag gga ccg gtg aag aag cct gtc gct
ttg aaa gtg aaa gct aag 4478Tyr Glu Gly Pro Val Lys Lys Pro Val Ala
Leu Lys Val Lys Ala Lys 895 900
905aac ttg ata gtc act gag agt ggt gcc cca ccg acc gac ttg caa aag
4526Asn Leu Ile Val Thr Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys
910 915 920atg gtc atg ggc aac aca aag
cct gtt gag ctc atc ctt gac ggg aag 4574Met Val Met Gly Asn Thr Lys
Pro Val Glu Leu Ile Leu Asp Gly Lys 925 930
935aca gta gcc atc tgt tgt gct act gga gtg ttt ggc act gct tac ctc
4622Thr Val Ala Ile Cys Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu940
945 950 955gtg cct cgt cat
ctt ttc gca gag aag tat gac aag atc atg ctg gat 4670Val Pro Arg His
Leu Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp 960
965 970ggc aga gcc atg aca gac agt gac tac aga
gtg ttt gag ttt gag att 4718Gly Arg Ala Met Thr Asp Ser Asp Tyr Arg
Val Phe Glu Phe Glu Ile 975 980
985aaa gta aaa gga cag gac atg ctc tca gac gct gcg ctc atg gtg ctc
4766Lys Val Lys Gly Gln Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu
990 995 1000cac cgt ggg aac cgc gtg aga
gat atc acg aaa cac ttt cgt gat aca 4814His Arg Gly Asn Arg Val Arg
Asp Ile Thr Lys His Phe Arg Asp Thr 1005 1010
1015gca aga atg aag aaa ggc acc ccc gtc gtc ggt gtg gtc aac aac gcc
4862Ala Arg Met Lys Lys Gly Thr Pro Val Val Gly Val Val Asn Asn
Ala1020 1025 1030 1035gac gtt
ggg aga ctg att ttc tct ggt gag gcc ctc acc tac aag gat 4910Asp Val
Gly Arg Leu Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp
1040 1045 1050att gta gtg tgc atg gac gga
gac acc atg cct ggc ctc ttt gcc tac 4958Ile Val Val Cys Met Asp Gly
Asp Thr Met Pro Gly Leu Phe Ala Tyr 1055 1060
1065aaa gcc gcc acc aag gca ggc tac tgt gga gga gcc gtt ctc
gcc aag 5006Lys Ala Ala Thr Lys Ala Gly Tyr Cys Gly Gly Ala Val Leu
Ala Lys 1070 1075 1080gac ggg gcc
gac act ttc atc gtc ggc act cac tcc gca gga ggc aat 5054Asp Gly Ala
Asp Thr Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn 1085
1090 1095gga gtt gga tac tgc tca tgc gtt tcc agg tcc atg
ctt ctc aga atg 5102Gly Val Gly Tyr Cys Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met1100 1105 1110
1115aag gca cac gtt gac cct gaa cca caa cac gag tagtaatttt tctagaatcg
5155Lys Ala His Val Asp Pro Glu Pro Gln His Glu 1120
1125atcccgggtt tttatgacta gttaatcacg gccgcttata aagatctaaa
atgcataatt 5215tctaaataat gaaaaaaagt acatcatgag caacgcgtta gtatatttta
caatggagat 5275taacgctcta taccgttcta tgtttattga ttcagatgat gttttagaaa
agaaagttat 5335tgaatatgaa aactttaatg aagatgaaga tgacgacgat gattattgtt
gtaaatctgt 5395tttagatgaa gaagatgacg cgctaaagta tactatggtt acaaagtata
agtctatact 5455actaatggcg acttgtgcaa gaaggtatag tatagtgaaa atgttgttag
attatgatta 5515tgaaaaacca aataaatcag atccatatct aaaggtatct cctttgcaca
taatttcatc 5575tattcctagt ttagaatac
559471126PRTFoot-and-mouth disease virus 7Met Gly Ala Gly Gln
Ser Ser Pro Ala Thr Gly Ser Gln Asn Gln Ser1 5
10 15Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr
Met Gln Gln Tyr Gln 20 25
30Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly Gly Ser
35 40 45Asn Glu Gly Ser Thr Asp Thr Thr
Ser Thr His Thr Thr Asn Thr Gln 50 55
60Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu65
70 75 80Phe Gly Ala Leu Leu
Ala Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu 85
90 95Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His
Thr Thr Ser Thr Thr 100 105
110Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu Asp His
115 120 125Val Ala Gly Pro Asn Thr Ser
Gly Leu Glu Thr Arg Val Val Gln Ala 130 135
140Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys
Ala145 150 155 160Phe Gly
His Leu Glu Lys Leu Glu Leu Pro Ser Asp His His Gly Val
165 170 175Phe Gly His Leu Val Asp Ser
Tyr Ala Tyr Met Arg Asn Gly Trp Asp 180 185
190Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys
Leu Leu 195 200 205Val Ala Met Val
Pro Glu Trp Lys Glu Phe Asp Thr Arg Glu Lys Tyr 210
215 220Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro
Arg Thr Asn Met225 230 235
240Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln
245 250 255Tyr Lys Lys His Lys
Pro Trp Thr Leu Val Val Met Val Val Ser Pro 260
265 270Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys
Val Tyr Ala Asn 275 280 285Ile Ala
Pro Thr Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu 290
295 300Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr
Gly Gly Leu Val Thr305 310 315
320Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn Pro
325 330 335Pro Arg Thr Asn
Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala 340
345 350Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp
Gly Lys Pro Tyr Val 355 360 365Thr
Thr Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser 370
375 380Leu Ala Ala Lys His Met Ser Asn Thr Tyr
Leu Ser Gly Ile Ala Gln385 390 395
400Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe
Thr 405 410 415Gly Ser Thr
Asp Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro 420
425 430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu
Arg Ala Ala His Cys Ile 435 440
445His Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile 450
455 460Pro Tyr Val Ser Ala Ala Asp Tyr
Ala Tyr Thr Ala Ser Asp Thr Ala465 470
475 480Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr
Gln Ile Thr His 485 490
495Gly Lys Ala Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys
500 505 510Asp Phe Glu Leu Arg Leu
Pro Ile Asp Pro Arg Gln Gln Thr Thr Ala 515 520
525Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu Asn
Tyr Gly 530 535 540Gly Glu Thr Gln Ile
Gln Arg Arg His His Thr Asp Ile Gly Phe Ile545 550
555 560Met Asp Arg Phe Val Lys Ile Gln Ser Leu
Ser Pro Thr His Val Ile 565 570
575Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
580 585 590Ala Ala Thr Tyr Tyr
Phe Ser Asp Leu Glu Ile Val Val Arg His Glu 595
600 605Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu
Ser Ala Leu Leu 610 615 620Asn Thr Ser
Asn Pro Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu625
630 635 640Ala Leu Pro Tyr Thr Ala Pro
His Arg Val Leu Ala Thr Val Tyr Asn 645
650 655Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg
Arg Gly Asp Met 660 665 670Gly
Ser Leu Ala Ala Arg Val Val Lys Gln Leu Pro Ala Ser Phe Asn 675
680 685Tyr Gly Ala Ile Lys Ala Asp Ala Ile
His Glu Leu Leu Val Arg Met 690 695
700Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val705
710 715 720Ser Ser Gln Asp
Arg His Lys Gln Lys Ile Ile Ala Pro Ala Lys Gln 725
730 735Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala
Gly Asp Val Glu Ser Asn 740 745
750Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser Lys Leu
755 760 765Val Asp Thr Ile Asn Gln Met
Gln Glu Asp Met Ser Thr Lys His Gly 770 775
780Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr
Gly785 790 795 800Val Lys
Ala Ile Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr Lys
805 810 815Leu Ile Lys Leu Leu Ser Arg
Leu Ser Cys Met Ala Ala Val Ala Ala 820 825
830Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp
Thr Gly 835 840 845Leu Glu Arg Gln
Arg Pro Leu Lys Val Arg Ala Lys Leu Pro Gln Gln 850
855 860Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys
Pro Leu Lys Val865 870 875
880Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro Val
885 890 895Lys Lys Pro Val Ala
Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr 900
905 910Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met
Val Met Gly Asn 915 920 925Thr Lys
Pro Val Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys 930
935 940Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu
Val Pro Arg His Leu945 950 955
960Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met Thr
965 970 975Asp Ser Asp Tyr
Arg Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln 980
985 990Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu
His Arg Gly Asn Arg 995 1000 1005Val
Arg Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys 1010
1015 1020Gly Thr Pro Val Val Gly Val Val Asn Asn
Ala Asp Val Gly Arg Leu1025 1030 1035
1040Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys
Met 1045 1050 1055Asp Gly
Asp Thr Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys 1060
1065 1070Ala Gly Tyr Cys Gly Gly Ala Val Leu
Ala Lys Asp Gly Ala Asp Thr 1075 1080
1085Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val Gly Tyr Cys
1090 1095 1100Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met Lys Ala His Val Asp1105 1110
1115 1120Pro Glu Pro Gln His Glu
112586442DNAFoot-and-mouth disease virusCDS(1640)..(5017) 8gaccctttac
aagaataaaa gaagaaacaa ctgtgaaata gtttataaat gtaattcgta 60tgcagaaaac
gataatatat tttggtatga gaaatctaaa ggagacatag tttgtataga 120catgcgctct
tccgatgaga tattcgatgc ttttctaatg tatcatatag ctacaagata 180tgcctatcat
gatgatgata tatatctaca aatagtgtta tattattcta ataatcaaaa 240tgttatatct
tatattacga aaaataaata cgttaagtat ataagaaata aaactagaga 300cgatattcat
aaagtaaaaa tattagctct agaagacttt acaacggaag aaatatattg 360ttggattagt
aatatataac agcgtagctg cacggttttg atcattttcc aacaatataa 420accaatgaag
gaggacgact catcaaacat aaataacatt cacggaaaat attcagtatc 480agatttatca
caagatgatt atgttattga atgtatagac ggatcttttg attcgatcaa 540gtatagagat
ataaaggtta taataatgaa gaataacggt tacgttaatt gtagtaaatt 600atgtaaaatg
cggaataaat acttttctag atggttgcgt ctttctactt ctaaagcatt 660attagacatt
tacaataata agtcagtaga taatgctatt gttaaagtct atggtaaagg 720taagaaactt
attataacag gattttatct caaacaaaat atgatacgtt atgttattga 780gtggataggg
gatgatttta caaacgatat atacaaaatg attaatttct ataatgcgtt 840attcggtaac
gatgaattaa aaatagtatc ctgtgaaaac actctatgcc cgtttataga 900acttggtaga
tgctattatg gtaaaaaatg taagtatata cacggagatc aatgtgatat 960ctgtggtcta
tatatactac accctaccga tattaaccaa cgagtttctc acaagaaaac 1020ttgtttagta
gatagagatt ctttgattgt gtttaaaaga agtaccagta aaaagtgtgg 1080catatgcata
gaagaaataa acaaaaaaca tatttccgaa cagtattttg gaattctccc 1140aagttgtaaa
catatttttt gcctatcatg tataagacgt tgggcagata ctaccagaaa 1200tacagatact
gaaaatacgt gtcctgaatg tagaatagtt tttcctttca taatacccag 1260taggtattgg
atagataata aatatgataa aaaaatatta tataatagat ataagaaaat 1320gatttttaca
aaaataccta taagaacaat aaaaatataa ttacatttac ggaaaatagc 1380tggttttagt
ttaccaactt agagtaatta tcatattgaa tctatattgc taattagcta 1440ataaaaaccc
gggttaatta attagtcatc aggcagggcg agaacgagac tatctgctcg 1500ttaattaatt
agagcttctt tattctatac ttaaaaagtg aaaataaata caaaggttct 1560tgagggttgt
gttaaattga aagcgagaaa taatcataaa ttatttcatt atcgcgatat 1620ccgttaagtt
tgtatcgta atg gga gct ggg caa tcc agc cca gca acc ggc 1672
Met Gly Ala Gly Gln Ser Ser Pro Ala Thr Gly 1
5 10tcg cag aac cag tct ggc aac act ggc
agc ata atc aac aac tac tac 1720Ser Gln Asn Gln Ser Gly Asn Thr Gly
Ser Ile Ile Asn Asn Tyr Tyr 15 20
25atg caa cag tac cag aac tcc atg gac aca cag ttg gga gac aat gcc
1768Met Gln Gln Tyr Gln Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala
30 35 40atc agt gga ggc tcc aac gag
ggc tcc acg gac aca act tca aca cac 1816Ile Ser Gly Gly Ser Asn Glu
Gly Ser Thr Asp Thr Thr Ser Thr His 45 50
55aca acc aac act caa aac aat gac tgg ttc tcg aag ctc gcc agt tca
1864Thr Thr Asn Thr Gln Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser60
65 70 75gct ttt acc ggt
ctg ttc ggt gca ctg ctc gcc gac aag aag aca gag 1912Ala Phe Thr Gly
Leu Phe Gly Ala Leu Leu Ala Asp Lys Lys Thr Glu 80
85 90gaa acg aca ctt ctt gag gac cgc atc ctc
acc acc cgc aac ggg cac 1960Glu Thr Thr Leu Leu Glu Asp Arg Ile Leu
Thr Thr Arg Asn Gly His 95 100
105acc acc tcg acg acc caa tcg agt gtg ggt gtc aca cac ggg tac tcc
2008Thr Thr Ser Thr Thr Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser
110 115 120aca gag gag gac cac gtt gct
ggg ccc aac aca tcg ggc ctg gag acg 2056Thr Glu Glu Asp His Val Ala
Gly Pro Asn Thr Ser Gly Leu Glu Thr 125 130
135cga gtg gtg cag gca gag aga ttc tac aaa aag tac ttg ttt gac tgg
2104Arg Val Val Gln Ala Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp140
145 150 155aca acg gac aag
gca ttt gga cac ctg gaa aag ctg gag ctc ccg tcc 2152Thr Thr Asp Lys
Ala Phe Gly His Leu Glu Lys Leu Glu Leu Pro Ser 160
165 170gac cac cac ggt gtc ttt gga cac ttg gtg
gac tcg tac gcc tat atg 2200Asp His His Gly Val Phe Gly His Leu Val
Asp Ser Tyr Ala Tyr Met 175 180
185aga aat ggc tgg gat gtt gag gtg tcc gct gtt ggc aac cag ttc aac
2248Arg Asn Gly Trp Asp Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn
190 195 200ggc ggg tgc ctc ctg gtg gcc
atg gta cct gaa tgg aag gaa ttt gac 2296Gly Gly Cys Leu Leu Val Ala
Met Val Pro Glu Trp Lys Glu Phe Asp 205 210
215aca cgg gag aaa tac caa ctc acc ctt ttc ccg cac cag ttt att agc
2344Thr Arg Glu Lys Tyr Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser220
225 230 235ccc aga act aac
atg act gcc cac atc acg gtc ccc tac ctt ggt gtg 2392Pro Arg Thr Asn
Met Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val 240
245 250aac agg tat gat cag tac aag aag cat aag
ccc tgg aca ttg gtt gtc 2440Asn Arg Tyr Asp Gln Tyr Lys Lys His Lys
Pro Trp Thr Leu Val Val 255 260
265atg gtc gtg tcg cca ctt acg gtc aac aac act agt gcg gca caa atc
2488Met Val Val Ser Pro Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile
270 275 280aag gtc tac gcc aac ata gct
ccg acc tat gtt cac gtg gcc ggt gaa 2536Lys Val Tyr Ala Asn Ile Ala
Pro Thr Tyr Val His Val Ala Gly Glu 285 290
295ctc ccc tcg aaa gag ggg att ttc ccg gtt gca tgt gcg gac ggt tac
2584Leu Pro Ser Lys Glu Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr300
305 310 315gga gga ttg gtg
acg aca gac ccg aag aca gct gac cct gct tat ggc 2632Gly Gly Leu Val
Thr Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly 320
325 330aag gtg tac aac ccg cct agg act aac tac
cct ggg cgc ttc acc aac 2680Lys Val Tyr Asn Pro Pro Arg Thr Asn Tyr
Pro Gly Arg Phe Thr Asn 335 340
345ctg ttg gac gtg gcc gaa gcg tgt ccc act ttc ctc tgc ttt gac gac
2728Leu Leu Asp Val Ala Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp
350 355 360ggg aaa ccg tac gtc acc acg
cgg acg gat gac acc cga ctt ttg gcc 2776Gly Lys Pro Tyr Val Thr Thr
Arg Thr Asp Asp Thr Arg Leu Leu Ala 365 370
375aag ttt gac ctt tcc ctt gcc gca aaa cat atg tcc aac aca tac ctg
2824Lys Phe Asp Leu Ser Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu380
385 390 395tca ggg att gct
cag tac tac aca cag tac tct ggc acc atc aat ttg 2872Ser Gly Ile Ala
Gln Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu 400
405 410cat ttc atg ttt aca ggt tcc act gat tca
aag gcc cga tac atg gtg 2920His Phe Met Phe Thr Gly Ser Thr Asp Ser
Lys Ala Arg Tyr Met Val 415 420
425gcc tac atc cca cct ggg gtg gag aca cca ccg gac aca cct gaa agg
2968Ala Tyr Ile Pro Pro Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg
430 435 440gct gcc cac tgc att cac gct
gaa tgg gac act gga cta aac tcc aaa 3016Ala Ala His Cys Ile His Ala
Glu Trp Asp Thr Gly Leu Asn Ser Lys 445 450
455ttc act ttc tca atc ccg tac gta tcc gcc gcg gat tac gcg tac aca
3064Phe Thr Phe Ser Ile Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr Thr460
465 470 475gcg tct gac acg
gca gaa aca atc aac gta cag gga tgg gtc tgc atc 3112Ala Ser Asp Thr
Ala Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile 480
485 490tac caa att aca cac ggg aag gct gaa aat
gac acc ttg gtc gtg tcg 3160Tyr Gln Ile Thr His Gly Lys Ala Glu Asn
Asp Thr Leu Val Val Ser 495 500
505gtt agc gcc ggc aaa gac ttt gag ttg cgc ctc ccg att gac ccc cgc
3208Val Ser Ala Gly Lys Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg
510 515 520cag cag acc acc gct acc ggg
gaa tca gca gac ccg gtc acc acc acc 3256Gln Gln Thr Thr Ala Thr Gly
Glu Ser Ala Asp Pro Val Thr Thr Thr 525 530
535gtg gag aac tac ggc ggt gag aca caa atc cag aga cgt cac cac acg
3304Val Glu Asn Tyr Gly Gly Glu Thr Gln Ile Gln Arg Arg His His Thr540
545 550 555gac att ggt ttc
atc atg gac aga ttt gtg aag atc caa agc ttg agc 3352Asp Ile Gly Phe
Ile Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser 560
565 570cca aca cat gtc att gac ctc atg cag gct
cac caa cac ggt ctg gtg 3400Pro Thr His Val Ile Asp Leu Met Gln Ala
His Gln His Gly Leu Val 575 580
585ggt gcc ttg ctg cgt gca gcc acg tac tac ttt tct gac ctg gaa att
3448Gly Ala Leu Leu Arg Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile
590 595 600gtt gta cgg cac gaa ggc aat
ctg acc tgg gtg ccc aac ggc gcc cct 3496Val Val Arg His Glu Gly Asn
Leu Thr Trp Val Pro Asn Gly Ala Pro 605 610
615gaa tca gcc ctg ttg aac acc agc aac ccc act gcc tac aac aag gca
3544Glu Ser Ala Leu Leu Asn Thr Ser Asn Pro Thr Ala Tyr Asn Lys Ala620
625 630 635cca ttc acg aga
ctc gct ctc ccc tac act gcg ccg cac cgt gtg ctg 3592Pro Phe Thr Arg
Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu 640
645 650gca aca gtg tac aac ggg acg agt aag tat
gct gtg ggt ggt tca ggc 3640Ala Thr Val Tyr Asn Gly Thr Ser Lys Tyr
Ala Val Gly Gly Ser Gly 655 660
665aga aga ggc gac atg ggg tct ctc gcg gcg cga gtc gtg aaa cag ctt
3688Arg Arg Gly Asp Met Gly Ser Leu Ala Ala Arg Val Val Lys Gln Leu
670 675 680cct gct tca ttt aac tac ggt
gca atc aag gcc gac gcc atc cac gaa 3736Pro Ala Ser Phe Asn Tyr Gly
Ala Ile Lys Ala Asp Ala Ile His Glu 685 690
695ctt ctc gtg cgc atg aaa cgg gcc gag ctc tac tgc ccc aga ccg ctg
3784Leu Leu Val Arg Met Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu700
705 710 715ttg gca ata gag
gtg tct tcg caa gac agg cac aag caa aag atc att 3832Leu Ala Ile Glu
Val Ser Ser Gln Asp Arg His Lys Gln Lys Ile Ile 720
725 730gca cca gca aag cag ctt ctg aat ttt gac
ctg ctc aag ttg gcc gga 3880Ala Pro Ala Lys Gln Leu Leu Asn Phe Asp
Leu Leu Lys Leu Ala Gly 735 740
745gac gtt gag tcc aac ccc ggg cca ttc ttc ttt gct gac gtt agg tca
3928Asp Val Glu Ser Asn Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser
750 755 760aac ttt tca aag ttg gta gac
aca atc aac cag atg cag gag gac atg 3976Asn Phe Ser Lys Leu Val Asp
Thr Ile Asn Gln Met Gln Glu Asp Met 765 770
775tcc aca aaa cac ggg ccc gac ttc aac cgg ttg gtg tcc gca ttt gag
4024Ser Thr Lys His Gly Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu780
785 790 795gaa ttg gcc act
gga gtt aaa gct atc agg acc ggt ctc gac gag gcc 4072Glu Leu Ala Thr
Gly Val Lys Ala Ile Arg Thr Gly Leu Asp Glu Ala 800
805 810aaa ccc tgg tac aag ctt atc aaa ctc cta
agc cgc ctg tcg tgc atg 4120Lys Pro Trp Tyr Lys Leu Ile Lys Leu Leu
Ser Arg Leu Ser Cys Met 815 820
825gcc gct gtg gca gca cgg tcc aag gac cca gtc ctt gtg gcc atc atg
4168Ala Ala Val Ala Ala Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met
830 835 840ctg gcc gac acc ggt ctc gag
cgt cag aga cct ctg aaa gtg aga gct 4216Leu Ala Asp Thr Gly Leu Glu
Arg Gln Arg Pro Leu Lys Val Arg Ala 845 850
855aag ctc cca cag cag gaa gga cct tac gct ggc ccg ttg gag aga cag
4264Lys Leu Pro Gln Gln Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln860
865 870 875aaa ccg ctg aaa
gtg aaa gca aaa gcc ccg gtc gtc aag gaa gga cct 4312Lys Pro Leu Lys
Val Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro 880
885 890tac gag gga ccg gtg aag aag cct gtc gct
ttg aaa gtg aaa gct aag 4360Tyr Glu Gly Pro Val Lys Lys Pro Val Ala
Leu Lys Val Lys Ala Lys 895 900
905aac ttg ata gtc act gag agt ggt gcc cca ccg acc gac ttg caa aag
4408Asn Leu Ile Val Thr Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys
910 915 920atg gtc atg ggc aac aca aag
cct gtt gag ctc atc ctt gac ggg aag 4456Met Val Met Gly Asn Thr Lys
Pro Val Glu Leu Ile Leu Asp Gly Lys 925 930
935aca gta gcc atc tgt tgt gct act gga gtg ttt ggc act gct tac ctc
4504Thr Val Ala Ile Cys Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu940
945 950 955gtg cct cgt cat
ctt ttc gca gag aag tat gac aag atc atg ctg gat 4552Val Pro Arg His
Leu Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp 960
965 970ggc aga gcc atg aca gac agt gac tac aga
gtg ttt gag ttt gag att 4600Gly Arg Ala Met Thr Asp Ser Asp Tyr Arg
Val Phe Glu Phe Glu Ile 975 980
985aaa gta aaa gga cag gac atg ctc tca gac gct gcg ctc atg gtg ctc
4648Lys Val Lys Gly Gln Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu
990 995 1000cac cgt ggg aac cgc gtg aga
gat atc acg aaa cac ttt cgt gat aca 4696His Arg Gly Asn Arg Val Arg
Asp Ile Thr Lys His Phe Arg Asp Thr 1005 1010
1015gca aga atg aag aaa ggc acc ccc gtc gtc ggt gtg gtc aac aac gcc
4744Ala Arg Met Lys Lys Gly Thr Pro Val Val Gly Val Val Asn Asn
Ala1020 1025 1030 1035gac gtt
ggg aga ctg att ttc tct ggt gag gcc ctc acc tac aag gat 4792Asp Val
Gly Arg Leu Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp
1040 1045 1050att gta gtg tgc atg gac gga
gac acc atg cct ggc ctc ttt gcc tac 4840Ile Val Val Cys Met Asp Gly
Asp Thr Met Pro Gly Leu Phe Ala Tyr 1055 1060
1065aaa gcc gcc acc aag gca ggc tac tgt gga gga gcc gtt ctc
gcc aag 4888Lys Ala Ala Thr Lys Ala Gly Tyr Cys Gly Gly Ala Val Leu
Ala Lys 1070 1075 1080gac ggg gcc
gac act ttc atc gtc ggc act cac tcc gca gga ggc aat 4936Asp Gly Ala
Asp Thr Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn 1085
1090 1095gga gtt gga tac tgc tca tgc gtt tcc agg tcc atg
ctt ctc aga atg 4984Gly Val Gly Tyr Cys Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met1100 1105 1110
1115aag gca cac gtt gac cct gaa cca caa cac gag tagtaatttt tctagaggat
5037Lys Ala His Val Asp Pro Glu Pro Gln His Glu 1120
1125ccctcgagtt tttattgact agttaatcat aagataaata atatacagca
ttgtaaccat 5097cgtcatccgt tatacgggga ataatattac catacagtat tattaaattt
tcttacgaag 5157aatatagatc ggtatttatc gttagtttat tttacattta ttaattaaac
atgtctacta 5217ttacctgtta tggaaatgac aaatttagtt atataattta tgataaaatt
aagataataa 5277taatgaaatc aaataattat gtaaatgcta ctagattatg tgaattacga
ggaagaaagt 5337ttacgaactg gaaaaaatta agtgaatcta aaatattagt cgataatgta
aaaaaaataa 5397atgataaaac taaccagtta aaaacggata tgattatata cgttaaggat
attgatcata 5457aaggaagaga tacttgcggt tactatgtac accaagatct ggtatcttct
atatcaaatt 5517ggatatctcc gttattcgcc gttaaggtaa ataaaattat taactattat
atatgtaatg 5577aatatgatat acgacttagc gaaatggaat ctgatatgac agaagtaata
gatgtagttg 5637ataaattagt aggaggatac aatgatgaaa tagcagaaat aatatatttg
tttaataaat 5697ttatagaaaa atatattgct aacatatcgt tatcaactga attatctagt
atattaaata 5757attttataaa ttttaataaa aaatacaata acgacataaa agatattaaa
tctttaattc 5817ttgatctgaa aaacacatct ataaaactag ataaaaagtt attcgataaa
gataataatg 5877aatcgaacga tgaaaaattg gaaacagaag ttgataagct aatttttttc
atctaaatag 5937tattatttta ttgaagtacg aagttttacg ttagataaat aataaaggtc
gatttttatt 5997ttgttaaata tcaaatatgt cattatctga taaagataca aaaacacacg
gtgattatca 6057accatctaac gaacagatat tacaaaaaat acgtcggact atggaaaacg
aagctgatag 6117cctcaataga agaagcatta aagaaattgt tgtagatgtt atgaagaatt
gggatcatcc 6177tctcaacgaa gaaatagata aagttctaaa ctggaaaaat gatacattaa
acgatttaga 6237tcatctaaat acagatgata atattaagga aatcatacaa tgtctgatta
gagaatttgc 6297gtttaaaaag atcaattcta ttatgtatag ttatgctatg gtaaaactca
attcagataa 6357cgaaacattg aaagataaaa ttaaggatta ttttatagaa actattctta
aagacaaacg 6417tggttataaa caaaagccat taccc
644291126PRTFoot-and-mouth disease virus 9Met Gly Ala Gly Gln
Ser Ser Pro Ala Thr Gly Ser Gln Asn Gln Ser1 5
10 15Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr
Met Gln Gln Tyr Gln 20 25
30Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly Gly Ser
35 40 45Asn Glu Gly Ser Thr Asp Thr Thr
Ser Thr His Thr Thr Asn Thr Gln 50 55
60Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu65
70 75 80Phe Gly Ala Leu Leu
Ala Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu 85
90 95Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His
Thr Thr Ser Thr Thr 100 105
110Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu Asp His
115 120 125Val Ala Gly Pro Asn Thr Ser
Gly Leu Glu Thr Arg Val Val Gln Ala 130 135
140Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys
Ala145 150 155 160Phe Gly
His Leu Glu Lys Leu Glu Leu Pro Ser Asp His His Gly Val
165 170 175Phe Gly His Leu Val Asp Ser
Tyr Ala Tyr Met Arg Asn Gly Trp Asp 180 185
190Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys
Leu Leu 195 200 205Val Ala Met Val
Pro Glu Trp Lys Glu Phe Asp Thr Arg Glu Lys Tyr 210
215 220Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro
Arg Thr Asn Met225 230 235
240Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln
245 250 255Tyr Lys Lys His Lys
Pro Trp Thr Leu Val Val Met Val Val Ser Pro 260
265 270Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys
Val Tyr Ala Asn 275 280 285Ile Ala
Pro Thr Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu 290
295 300Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr
Gly Gly Leu Val Thr305 310 315
320Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn Pro
325 330 335Pro Arg Thr Asn
Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala 340
345 350Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp
Gly Lys Pro Tyr Val 355 360 365Thr
Thr Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser 370
375 380Leu Ala Ala Lys His Met Ser Asn Thr Tyr
Leu Ser Gly Ile Ala Gln385 390 395
400Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe
Thr 405 410 415Gly Ser Thr
Asp Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro 420
425 430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu
Arg Ala Ala His Cys Ile 435 440
445His Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile 450
455 460Pro Tyr Val Ser Ala Ala Asp Tyr
Ala Tyr Thr Ala Ser Asp Thr Ala465 470
475 480Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr
Gln Ile Thr His 485 490
495Gly Lys Ala Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys
500 505 510Asp Phe Glu Leu Arg Leu
Pro Ile Asp Pro Arg Gln Gln Thr Thr Ala 515 520
525Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu Asn
Tyr Gly 530 535 540Gly Glu Thr Gln Ile
Gln Arg Arg His His Thr Asp Ile Gly Phe Ile545 550
555 560Met Asp Arg Phe Val Lys Ile Gln Ser Leu
Ser Pro Thr His Val Ile 565 570
575Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
580 585 590Ala Ala Thr Tyr Tyr
Phe Ser Asp Leu Glu Ile Val Val Arg His Glu 595
600 605Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu
Ser Ala Leu Leu 610 615 620Asn Thr Ser
Asn Pro Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu625
630 635 640Ala Leu Pro Tyr Thr Ala Pro
His Arg Val Leu Ala Thr Val Tyr Asn 645
650 655Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg
Arg Gly Asp Met 660 665 670Gly
Ser Leu Ala Ala Arg Val Val Lys Gln Leu Pro Ala Ser Phe Asn 675
680 685Tyr Gly Ala Ile Lys Ala Asp Ala Ile
His Glu Leu Leu Val Arg Met 690 695
700Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val705
710 715 720Ser Ser Gln Asp
Arg His Lys Gln Lys Ile Ile Ala Pro Ala Lys Gln 725
730 735Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala
Gly Asp Val Glu Ser Asn 740 745
750Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser Lys Leu
755 760 765Val Asp Thr Ile Asn Gln Met
Gln Glu Asp Met Ser Thr Lys His Gly 770 775
780Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr
Gly785 790 795 800Val Lys
Ala Ile Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr Lys
805 810 815Leu Ile Lys Leu Leu Ser Arg
Leu Ser Cys Met Ala Ala Val Ala Ala 820 825
830Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp
Thr Gly 835 840 845Leu Glu Arg Gln
Arg Pro Leu Lys Val Arg Ala Lys Leu Pro Gln Gln 850
855 860Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys
Pro Leu Lys Val865 870 875
880Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro Val
885 890 895Lys Lys Pro Val Ala
Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr 900
905 910Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met
Val Met Gly Asn 915 920 925Thr Lys
Pro Val Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys 930
935 940Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu
Val Pro Arg His Leu945 950 955
960Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met Thr
965 970 975Asp Ser Asp Tyr
Arg Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln 980
985 990Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu
His Arg Gly Asn Arg 995 1000 1005Val
Arg Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys 1010
1015 1020Gly Thr Pro Val Val Gly Val Val Asn Asn
Ala Asp Val Gly Arg Leu1025 1030 1035
1040Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys
Met 1045 1050 1055Asp Gly
Asp Thr Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys 1060
1065 1070Ala Gly Tyr Cys Gly Gly Ala Val Leu
Ala Lys Asp Gly Ala Asp Thr 1075 1080
1085Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val Gly Tyr Cys
1090 1095 1100Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met Lys Ala His Val Asp1105 1110
1115 1120Pro Glu Pro Gln His Glu
11251024DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 10cgacaccggt ctcgagcgtc agag
241141DNAArtificial SequenceDescription of Artificial
Sequence Synthetic primer 11gttgaccctg aaccacaaca cgagtagtaa
tttttctgca g 411241DNAArtificial
SequenceDescription of Artificial Sequence Synthetic primer
12ctgcagaaaa attactactc gtgttgtggt tcagggtcaa c
41138PRTArtificial SequenceDescription of Artificial Sequence Synthetic
peptide 13Asp Thr Gly Leu Glu Arg Gln Arg1
5148PRTArtificial SequenceDescription of Artificial Sequence Synthetic
peptide 14Val Asp Pro Glu Pro Gln His Glu1
5155102DNAFoot-and-mouth disease virusCDS(1311)..(4688) 15ttcataaata
caagtttgat taaacttaag ttgttctaaa gttctttcct ccgaaggtat 60agaacaaagt
atttcttcta catccttact atttattgca gcttttaaca gcctatcacg 120tatcctattt
ttagtattgg tagaacgttt tagttctaaa gttaaaatat tagacataat 180tggcatattg
cttattcctt gcatagttga gtctgtagat cgtttcagta tatcactgat 240taatgtacta
ctgttatgat gaaatataga atcgatattg gcatttaact gttttgttat 300actaagtcta
gattttaaat cttctagtaa tatgctattt aatataaaag cttccacgtt 360tttgtataca
tttctttcca tattagtagc tactactaaa tgattatctt ctttcatatc 420ttgtagataa
gatagactat ctttatcttt attagtagaa aatacttctg gccatacatc 480gttaaatttt
tttgttgttg ttagatataa tattaaatat ctagaggatc ctattatttg 540tggtaaaatg
tttatagagt aaaatgatct ggctattaaa cataggccag ttaccataga 600atgctgcttc
ccgttacagt gttttaccat aaccatagat ctgcctgtat tgttgataca 660tataacagct
gtaaatccta aaaaattcct atcataatta ttaatattag gtaattcatt 720tccatgtgaa
agatagacta attttatatc ctttacctcc aaataattat ttacatctct 780taaacaatct
attttaatat cattaactgg tattttataa tatccagaaa ggtttgaagg 840ggttgatgga
ataagtctat taacatcgtt aagtaaatta ttaatatcat gaatctttat 900tatattatac
ccataagtta aatttatatt tactttctca tcatctgact tagttagttt 960gtaataaggt
gtgtctgaaa aaattaaaag gtaattcgtt gaatgaagct gtatttgctg 1020tatcattttt
atctaatttt ggagatttag cagtacttac ttcattagaa gaagaatctg 1080ccagttcctg
tctattactg atatttcgtt tcattattat atgatttata ttttactttt 1140tcaattatat
atactcattt gactagttaa tcaataaaaa gaattgttct ttattctata 1200cttaaaaagt
gaaaataaat acaaaggttc ttgagggttg tgttaaattg aaagcgagaa 1260ataatcataa
attatttcat tatcgcgata tccgttaagt ttgtatcgta atg gga 1316
Met Gly
1gct ggg caa tcc agc cca gca acc ggc tcg
cag aac cag tct ggc aac 1364Ala Gly Gln Ser Ser Pro Ala Thr Gly Ser
Gln Asn Gln Ser Gly Asn 5 10
15act ggc agc ata atc aac aac tac tac atg caa cag tac cag aac tcc
1412Thr Gly Ser Ile Ile Asn Asn Tyr Tyr Met Gln Gln Tyr Gln Asn Ser 20
25 30atg gac aca cag ttg gga gac aat
gcc atc agt gga ggc tcc aac gag 1460Met Asp Thr Gln Leu Gly Asp Asn
Ala Ile Ser Gly Gly Ser Asn Glu35 40 45
50ggc tcc acg gac aca act tca aca cac aca acc aac act
caa aac aat 1508Gly Ser Thr Asp Thr Thr Ser Thr His Thr Thr Asn Thr
Gln Asn Asn 55 60 65gac
tgg ttc tcg aag ctc gcc agt tca gct ttt acc ggt ctg ttc ggt 1556Asp
Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu Phe Gly 70
75 80gca ctg ctc gcc gac aag aag aca
gag gaa acg aca ctt ctt gag gac 1604Ala Leu Leu Ala Asp Lys Lys Thr
Glu Glu Thr Thr Leu Leu Glu Asp 85 90
95cgc atc ctc acc acc cgc aac ggg cac acc acc tcg acg acc caa tcg
1652Arg Ile Leu Thr Thr Arg Asn Gly His Thr Thr Ser Thr Thr Gln Ser
100 105 110agt gtg ggt gtc aca cac ggg
tac tcc aca gag gag gac cac gtt gct 1700Ser Val Gly Val Thr His Gly
Tyr Ser Thr Glu Glu Asp His Val Ala115 120
125 130ggg ccc aac aca tcg ggc ctg gag acg cga gtg gtg
cag gca gag aga 1748Gly Pro Asn Thr Ser Gly Leu Glu Thr Arg Val Val
Gln Ala Glu Arg 135 140
145ttc tac aaa aag tac ttg ttt gac tgg aca acg gac aag gca ttt gga
1796Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys Ala Phe Gly
150 155 160cac ctg gaa aag ctg gag
ctc ccg tcc gac cac cac ggt gtc ttt gga 1844His Leu Glu Lys Leu Glu
Leu Pro Ser Asp His His Gly Val Phe Gly 165 170
175cac ttg gtg gac tcg tac gcc tat atg aga aat ggc tgg gat
gtt gag 1892His Leu Val Asp Ser Tyr Ala Tyr Met Arg Asn Gly Trp Asp
Val Glu 180 185 190gtg tcc gct gtt ggc
aac cag ttc aac ggc ggg tgc ctc ctg gtg gcc 1940Val Ser Ala Val Gly
Asn Gln Phe Asn Gly Gly Cys Leu Leu Val Ala195 200
205 210atg gta cct gaa tgg aag gaa ttt gac aca
cgg gag aaa tac caa ctc 1988Met Val Pro Glu Trp Lys Glu Phe Asp Thr
Arg Glu Lys Tyr Gln Leu 215 220
225acc ctt ttc ccg cac cag ttt att agc ccc aga act aac atg act gcc
2036Thr Leu Phe Pro His Gln Phe Ile Ser Pro Arg Thr Asn Met Thr Ala
230 235 240cac atc acg gtc ccc tac
ctt ggt gtg aac agg tat gat cag tac aag 2084His Ile Thr Val Pro Tyr
Leu Gly Val Asn Arg Tyr Asp Gln Tyr Lys 245 250
255aag cat aag ccc tgg aca ttg gtt gtc atg gtc gtg tcg cca
ctt acg 2132Lys His Lys Pro Trp Thr Leu Val Val Met Val Val Ser Pro
Leu Thr 260 265 270gtc aac aac act agt
gcg gca caa atc aag gtc tac gcc aac ata gct 2180Val Asn Asn Thr Ser
Ala Ala Gln Ile Lys Val Tyr Ala Asn Ile Ala275 280
285 290ccg acc tat gtt cac gtg gcc ggt gaa ctc
ccc tcg aaa gag ggg att 2228Pro Thr Tyr Val His Val Ala Gly Glu Leu
Pro Ser Lys Glu Gly Ile 295 300
305ttc ccg gtt gca tgt gcg gac ggt tac gga gga ttg gtg acg aca gac
2276Phe Pro Val Ala Cys Ala Asp Gly Tyr Gly Gly Leu Val Thr Thr Asp
310 315 320ccg aag aca gct gac cct
gct tat ggc aag gtg tac aac ccg cct agg 2324Pro Lys Thr Ala Asp Pro
Ala Tyr Gly Lys Val Tyr Asn Pro Pro Arg 325 330
335act aac tac cct ggg cgc ttc acc aac ctg ttg gac gtg gcc
gaa gcg 2372Thr Asn Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala
Glu Ala 340 345 350tgt ccc act ttc ctc
tgc ttt gac gac ggg aaa ccg tac gtc acc acg 2420Cys Pro Thr Phe Leu
Cys Phe Asp Asp Gly Lys Pro Tyr Val Thr Thr355 360
365 370cgg acg gat gac acc cga ctt ttg gcc aag
ttt gac ctt tcc ctt gcc 2468Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys
Phe Asp Leu Ser Leu Ala 375 380
385gca aaa cat atg tcc aac aca tac ctg tca ggg att gct cag tac tac
2516Ala Lys His Met Ser Asn Thr Tyr Leu Ser Gly Ile Ala Gln Tyr Tyr
390 395 400aca cag tac tct ggc acc
atc aat ttg cat ttc atg ttt aca ggt tcc 2564Thr Gln Tyr Ser Gly Thr
Ile Asn Leu His Phe Met Phe Thr Gly Ser 405 410
415act gat tca aag gcc cga tac atg gtg gcc tac atc cca cct
ggg gtg 2612Thr Asp Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro
Gly Val 420 425 430gag aca cca ccg gac
aca cct gaa agg gct gcc cac tgc att cac gct 2660Glu Thr Pro Pro Asp
Thr Pro Glu Arg Ala Ala His Cys Ile His Ala435 440
445 450gaa tgg gac act gga cta aac tcc aaa ttc
act ttc tca atc ccg tac 2708Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe
Thr Phe Ser Ile Pro Tyr 455 460
465gta tcc gcc gcg gat tac gcg tac aca gcg tct gac acg gca gaa aca
2756Val Ser Ala Ala Asp Tyr Ala Tyr Thr Ala Ser Asp Thr Ala Glu Thr
470 475 480atc aac gta cag gga tgg
gtc tgc atc tac caa att aca cac ggg aag 2804Ile Asn Val Gln Gly Trp
Val Cys Ile Tyr Gln Ile Thr His Gly Lys 485 490
495gct gaa aat gac acc ttg gtc gtg tcg gtt agc gcc ggc aaa
gac ttt 2852Ala Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys
Asp Phe 500 505 510gag ttg cgc ctc ccg
att gac ccc cgc cag cag acc acc gct acc ggg 2900Glu Leu Arg Leu Pro
Ile Asp Pro Arg Gln Gln Thr Thr Ala Thr Gly515 520
525 530gaa tca gca gac ccg gtc acc acc acc gtg
gag aac tac ggc ggt gag 2948Glu Ser Ala Asp Pro Val Thr Thr Thr Val
Glu Asn Tyr Gly Gly Glu 535 540
545aca caa atc cag aga cgt cac cac acg gac att ggt ttc atc atg gac
2996Thr Gln Ile Gln Arg Arg His His Thr Asp Ile Gly Phe Ile Met Asp
550 555 560aga ttt gtg aag atc caa
agc ttg agc cca aca cat gtc att gac ctc 3044Arg Phe Val Lys Ile Gln
Ser Leu Ser Pro Thr His Val Ile Asp Leu 565 570
575atg cag gct cac caa cac ggt ctg gtg ggt gcc ttg ctg cgt
gca gcc 3092Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
Ala Ala 580 585 590acg tac tac ttt tct
gac ctg gaa att gtt gta cgg cac gaa ggc aat 3140Thr Tyr Tyr Phe Ser
Asp Leu Glu Ile Val Val Arg His Glu Gly Asn595 600
605 610ctg acc tgg gtg ccc aac ggc gcc cct gaa
tca gcc ctg ttg aac acc 3188Leu Thr Trp Val Pro Asn Gly Ala Pro Glu
Ser Ala Leu Leu Asn Thr 615 620
625agc aac ccc act gcc tac aac aag gca cca ttc acg aga ctc gct ctc
3236Ser Asn Pro Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu Ala Leu
630 635 640ccc tac act gcg ccg cac
cgt gtg ctg gca aca gtg tac aac ggg acg 3284Pro Tyr Thr Ala Pro His
Arg Val Leu Ala Thr Val Tyr Asn Gly Thr 645 650
655agt aag tat gct gtg ggt ggt tca ggc aga aga ggc gac atg
ggg tct 3332Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly Asp Met
Gly Ser 660 665 670ctc gcg gcg cga gtc
gtg aaa cag ctt cct gct tca ttt aac tac ggt 3380Leu Ala Ala Arg Val
Val Lys Gln Leu Pro Ala Ser Phe Asn Tyr Gly675 680
685 690gca atc aag gcc gac gcc atc cac gaa ctt
ctc gtg cgc atg aaa cgg 3428Ala Ile Lys Ala Asp Ala Ile His Glu Leu
Leu Val Arg Met Lys Arg 695 700
705gcc gag ctc tac tgc ccc aga ccg ctg ttg gca ata gag gtg tct tcg
3476Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val Ser Ser
710 715 720caa gac agg cac aag caa
aag atc att gca cca gca aag cag ctt ctg 3524Gln Asp Arg His Lys Gln
Lys Ile Ile Ala Pro Ala Lys Gln Leu Leu 725 730
735aat ttt gac ctg ctc aag ttg gcc gga gac gtt gag tcc aac
ccc ggg 3572Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp Val Glu Ser Asn
Pro Gly 740 745 750cca ttc ttc ttt gct
gac gtt agg tca aac ttt tca aag ttg gta gac 3620Pro Phe Phe Phe Ala
Asp Val Arg Ser Asn Phe Ser Lys Leu Val Asp755 760
765 770aca atc aac cag atg cag gag gac atg tcc
aca aaa cac ggg ccc gac 3668Thr Ile Asn Gln Met Gln Glu Asp Met Ser
Thr Lys His Gly Pro Asp 775 780
785ttc aac cgg ttg gtg tcc gca ttt gag gaa ttg gcc act gga gtt aaa
3716Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr Gly Val Lys
790 795 800gct atc agg acc ggt ctc
gac gag gcc aaa ccc tgg tac aag ctt atc 3764Ala Ile Arg Thr Gly Leu
Asp Glu Ala Lys Pro Trp Tyr Lys Leu Ile 805 810
815aaa ctc cta agc cgc ctg tcg tgc atg gcc gct gtg gca gca
cgg tcc 3812Lys Leu Leu Ser Arg Leu Ser Cys Met Ala Ala Val Ala Ala
Arg Ser 820 825 830aag gac cca gtc ctt
gtg gcc atc atg ctg gcc gac acc ggt ctc gag 3860Lys Asp Pro Val Leu
Val Ala Ile Met Leu Ala Asp Thr Gly Leu Glu835 840
845 850cgt cag aga cct ctg aaa gtg aga gct aag
ctc cca cag cag gaa gga 3908Arg Gln Arg Pro Leu Lys Val Arg Ala Lys
Leu Pro Gln Gln Glu Gly 855 860
865cct tac gct ggc ccg ttg gag aga cag aaa ccg ctg aaa gtg aaa gca
3956Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys Pro Leu Lys Val Lys Ala
870 875 880aaa gcc ccg gtc gtc aag
gaa gga cct tac gag gga ccg gtg aag aag 4004Lys Ala Pro Val Val Lys
Glu Gly Pro Tyr Glu Gly Pro Val Lys Lys 885 890
895cct gtc gct ttg aaa gtg aaa gct aag aac ttg ata gtc act
gag agt 4052Pro Val Ala Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr
Glu Ser 900 905 910ggt gcc cca ccg acc
gac ttg caa aag atg gtc atg ggc aac aca aag 4100Gly Ala Pro Pro Thr
Asp Leu Gln Lys Met Val Met Gly Asn Thr Lys915 920
925 930cct gtt gag ctc atc ctt gac ggg aag aca
gta gcc atc tgt tgt gct 4148Pro Val Glu Leu Ile Leu Asp Gly Lys Thr
Val Ala Ile Cys Cys Ala 935 940
945act gga gtg ttt ggc act gct tac ctc gtg cct cgt cat ctt ttc gca
4196Thr Gly Val Phe Gly Thr Ala Tyr Leu Val Pro Arg His Leu Phe Ala
950 955 960gag aag tat gac aag atc
atg ctg gat ggc aga gcc atg aca gac agt 4244Glu Lys Tyr Asp Lys Ile
Met Leu Asp Gly Arg Ala Met Thr Asp Ser 965 970
975gac tac aga gtg ttt gag ttt gag att aaa gta aaa gga cag
gac atg 4292Asp Tyr Arg Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln
Asp Met 980 985 990ctc tca gac gct gcg
ctc atg gtg ctc cac cgt ggg aac cgc gtg aga 4340Leu Ser Asp Ala Ala
Leu Met Val Leu His Arg Gly Asn Arg Val Arg995 1000
1005 1010gat atc acg aaa cac ttt cgt gat aca gca
aga atg aag aaa ggc acc 4388Asp Ile Thr Lys His Phe Arg Asp Thr Ala
Arg Met Lys Lys Gly Thr 1015 1020
1025ccc gtc gtc ggt gtg gtc aac aac gcc gac gtt ggg aga ctg att ttc
4436Pro Val Val Gly Val Val Asn Asn Ala Asp Val Gly Arg Leu Ile Phe
1030 1035 1040tct ggt gag gcc ctc
acc tac aag gat att gta gtg tgc atg gac gga 4484Ser Gly Glu Ala Leu
Thr Tyr Lys Asp Ile Val Val Cys Met Asp Gly 1045
1050 1055gac acc atg cct ggc ctc ttt gcc tac aaa gcc gcc
acc aag gca ggc 4532Asp Thr Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala
Thr Lys Ala Gly 1060 1065 1070tac tgt
gga gga gcc gtt ctc gcc aag gac ggg gcc gac act ttc atc 4580Tyr Cys
Gly Gly Ala Val Leu Ala Lys Asp Gly Ala Asp Thr Phe Ile1075
1080 1085 1090gtc ggc act cac tcc gca gga
ggc aat gga gtt gga tac tgc tca tgc 4628Val Gly Thr His Ser Ala Gly
Gly Asn Gly Val Gly Tyr Cys Ser Cys 1095
1100 1105gtt tcc agg tcc atg ctt ctc aga atg aag gca cac
gtt gac cct gaa 4676Val Ser Arg Ser Met Leu Leu Arg Met Lys Ala His
Val Asp Pro Glu 1110 1115
1120cca caa cac gag tagtaatttt tctgcagccc gggtttttat agctaattag
4728Pro Gln His Glu 1125tcattttttc gtaagtaagt atttttattt
aatacttttt attgtactta tgttaaatat 4788aactgatgat aacaaaatcc attatgtatt
atttataact gtaatttctt tagcgtagtt 4848agatgtccaa tctctctcaa atacatcggc
tatcttttta gtgagatttt gatctatgca 4908gttgaaactt atgaacgcgt gatgattaaa
atgtgaaccg tccaaatttg cagtcattat 4968atgagcgtat ctattatcta ctatcatcat
ctttgagtta ttaatatcat ctactttaga 5028attgatagga aatatgaata cctttgtagt
aatatctata ctatctacac ctaactcatt 5088aagacttttg atag
5102161126PRTFoot-and-mouth disease
virus 16Met Gly Ala Gly Gln Ser Ser Pro Ala Thr Gly Ser Gln Asn Gln Ser1
5 10 15Gly Asn Thr Gly
Ser Ile Ile Asn Asn Tyr Tyr Met Gln Gln Tyr Gln 20
25 30Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala
Ile Ser Gly Gly Ser 35 40 45Asn
Glu Gly Ser Thr Asp Thr Thr Ser Thr His Thr Thr Asn Thr Gln 50
55 60Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser
Ser Ala Phe Thr Gly Leu65 70 75
80Phe Gly Ala Leu Leu Ala Asp Lys Lys Thr Glu Glu Thr Thr Leu
Leu 85 90 95Glu Asp Arg
Ile Leu Thr Thr Arg Asn Gly His Thr Thr Ser Thr Thr 100
105 110Gln Ser Ser Val Gly Val Thr His Gly Tyr
Ser Thr Glu Glu Asp His 115 120
125Val Ala Gly Pro Asn Thr Ser Gly Leu Glu Thr Arg Val Val Gln Ala 130
135 140Glu Arg Phe Tyr Lys Lys Tyr Leu
Phe Asp Trp Thr Thr Asp Lys Ala145 150
155 160Phe Gly His Leu Glu Lys Leu Glu Leu Pro Ser Asp
His His Gly Val 165 170
175Phe Gly His Leu Val Asp Ser Tyr Ala Tyr Met Arg Asn Gly Trp Asp
180 185 190Val Glu Val Ser Ala Val
Gly Asn Gln Phe Asn Gly Gly Cys Leu Leu 195 200
205Val Ala Met Val Pro Glu Trp Lys Glu Phe Asp Thr Arg Glu
Lys Tyr 210 215 220Gln Leu Thr Leu Phe
Pro His Gln Phe Ile Ser Pro Arg Thr Asn Met225 230
235 240Thr Ala His Ile Thr Val Pro Tyr Leu Gly
Val Asn Arg Tyr Asp Gln 245 250
255Tyr Lys Lys His Lys Pro Trp Thr Leu Val Val Met Val Val Ser Pro
260 265 270Leu Thr Val Asn Asn
Thr Ser Ala Ala Gln Ile Lys Val Tyr Ala Asn 275
280 285Ile Ala Pro Thr Tyr Val His Val Ala Gly Glu Leu
Pro Ser Lys Glu 290 295 300Gly Ile Phe
Pro Val Ala Cys Ala Asp Gly Tyr Gly Gly Leu Val Thr305
310 315 320Thr Asp Pro Lys Thr Ala Asp
Pro Ala Tyr Gly Lys Val Tyr Asn Pro 325
330 335Pro Arg Thr Asn Tyr Pro Gly Arg Phe Thr Asn Leu
Leu Asp Val Ala 340 345 350Glu
Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly Lys Pro Tyr Val 355
360 365Thr Thr Arg Thr Asp Asp Thr Arg Leu
Leu Ala Lys Phe Asp Leu Ser 370 375
380Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu Ser Gly Ile Ala Gln385
390 395 400Tyr Tyr Thr Gln
Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe Thr 405
410 415Gly Ser Thr Asp Ser Lys Ala Arg Tyr Met
Val Ala Tyr Ile Pro Pro 420 425
430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg Ala Ala His Cys Ile
435 440 445His Ala Glu Trp Asp Thr Gly
Leu Asn Ser Lys Phe Thr Phe Ser Ile 450 455
460Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr Thr Ala Ser Asp Thr
Ala465 470 475 480Glu Thr
Ile Asn Val Gln Gly Trp Val Cys Ile Tyr Gln Ile Thr His
485 490 495Gly Lys Ala Glu Asn Asp Thr
Leu Val Val Ser Val Ser Ala Gly Lys 500 505
510Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg Gln Gln Thr
Thr Ala 515 520 525Thr Gly Glu Ser
Ala Asp Pro Val Thr Thr Thr Val Glu Asn Tyr Gly 530
535 540Gly Glu Thr Gln Ile Gln Arg Arg His His Thr Asp
Ile Gly Phe Ile545 550 555
560Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser Pro Thr His Val Ile
565 570 575Asp Leu Met Gln Ala
His Gln His Gly Leu Val Gly Ala Leu Leu Arg 580
585 590Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile Val
Val Arg His Glu 595 600 605Gly Asn
Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ser Ala Leu Leu 610
615 620Asn Thr Ser Asn Pro Thr Ala Tyr Asn Lys Ala
Pro Phe Thr Arg Leu625 630 635
640Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala Thr Val Tyr Asn
645 650 655Gly Thr Ser Lys
Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly Asp Met 660
665 670Gly Ser Leu Ala Ala Arg Val Val Lys Gln Leu
Pro Ala Ser Phe Asn 675 680 685Tyr
Gly Ala Ile Lys Ala Asp Ala Ile His Glu Leu Leu Val Arg Met 690
695 700Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro
Leu Leu Ala Ile Glu Val705 710 715
720Ser Ser Gln Asp Arg His Lys Gln Lys Ile Ile Ala Pro Ala Lys
Gln 725 730 735Leu Leu Asn
Phe Asp Leu Leu Lys Leu Ala Gly Asp Val Glu Ser Asn 740
745 750Pro Gly Pro Phe Phe Phe Ala Asp Val Arg
Ser Asn Phe Ser Lys Leu 755 760
765Val Asp Thr Ile Asn Gln Met Gln Glu Asp Met Ser Thr Lys His Gly 770
775 780Pro Asp Phe Asn Arg Leu Val Ser
Ala Phe Glu Glu Leu Ala Thr Gly785 790
795 800Val Lys Ala Ile Arg Thr Gly Leu Asp Glu Ala Lys
Pro Trp Tyr Lys 805 810
815Leu Ile Lys Leu Leu Ser Arg Leu Ser Cys Met Ala Ala Val Ala Ala
820 825 830Arg Ser Lys Asp Pro Val
Leu Val Ala Ile Met Leu Ala Asp Thr Gly 835 840
845Leu Glu Arg Gln Arg Pro Leu Lys Val Arg Ala Lys Leu Pro
Gln Gln 850 855 860Glu Gly Pro Tyr Ala
Gly Pro Leu Glu Arg Gln Lys Pro Leu Lys Val865 870
875 880Lys Ala Lys Ala Pro Val Val Lys Glu Gly
Pro Tyr Glu Gly Pro Val 885 890
895Lys Lys Pro Val Ala Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr
900 905 910Glu Ser Gly Ala Pro
Pro Thr Asp Leu Gln Lys Met Val Met Gly Asn 915
920 925Thr Lys Pro Val Glu Leu Ile Leu Asp Gly Lys Thr
Val Ala Ile Cys 930 935 940Cys Ala Thr
Gly Val Phe Gly Thr Ala Tyr Leu Val Pro Arg His Leu945
950 955 960Phe Ala Glu Lys Tyr Asp Lys
Ile Met Leu Asp Gly Arg Ala Met Thr 965
970 975Asp Ser Asp Tyr Arg Val Phe Glu Phe Glu Ile Lys
Val Lys Gly Gln 980 985 990Asp
Met Leu Ser Asp Ala Ala Leu Met Val Leu His Arg Gly Asn Arg 995
1000 1005Val Arg Asp Ile Thr Lys His Phe Arg
Asp Thr Ala Arg Met Lys Lys 1010 1015
1020Gly Thr Pro Val Val Gly Val Val Asn Asn Ala Asp Val Gly Arg Leu1025
1030 1035 1040Ile Phe Ser Gly
Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys Met 1045
1050 1055Asp Gly Asp Thr Met Pro Gly Leu Phe Ala
Tyr Lys Ala Ala Thr Lys 1060 1065
1070Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys Asp Gly Ala Asp Thr
1075 1080 1085Phe Ile Val Gly Thr His Ser
Ala Gly Gly Asn Gly Val Gly Tyr Cys 1090 1095
1100Ser Cys Val Ser Arg Ser Met Leu Leu Arg Met Lys Ala His Val
Asp1105 1110 1115 1120Pro Glu
Pro Gln His Glu 112517124DNAVaccinia virus 17ttctttattc
tatacttaaa aagtgaaaat aaatacaaag gttcttgagg gttgtgttaa 60attgaaagcg
agaaataatc ataaattatt tcattatcgc gatatccgtt aagtttgtat 120cgta
1241847DNAVaccinia virus 18ttctttattc tatacttaaa aagtgcaaat aaatacaaag
gttcttg 471960DNAArtificial SequenceDescription of
Artificial Sequence Synthetic primer 19gaattcttct ttattctata
cttaaaaagt gcaaataaat acaaaggttc ttgatgggag 602022DNAArtificial
SequenceDescription of Artificial Sequence Synthetic primer
20cttgccgcaa aacatatgtc ca
222122DNAArtificial SequenceDescription of Artificial Sequence Synthetic
primer 21tggacatatg ttttgcggca ag
222225DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 22cttgatggga gctgggcaat ccagc
252325DNAArtificial SequenceDescription of Artificial
Sequence Synthetic primer 23gctggattgc ccagctccca tcaag
25247PRTArtificial SequenceDescription of
Artificial Sequence Synthetic peptide 24Leu Ala Ala Lys His Met Ser1
5257PRTArtificial SequenceDescription of Artificial
Sequence Synthetic peptide 25Met Gly Ala Gly Gln Ser Ser1
5265025DNAFoot-and-mouth disease virusCDS(1234)..(4611) 26ttcataaata
caagtttgat taaacttaag ttgttctaaa gttctttcct ccgaaggtat 60agaacaaagt
atttcttcta catccttact atttattgca gcttttaaca gcctatcacg 120tatcctattt
ttagtattgg tagaacgttt tagttctaaa gttaaaatat tagacataat 180tggcatattg
cttattcctt gcatagttga gtctgtagat cgtttcagta tatcactgat 240taatgtacta
ctgttatgat gaaatataga atcgatattg gcatttaact gttttgttat 300actaagtcta
gattttaaat cttctagtaa tatgctattt aatataaaag cttccacgtt 360tttgtataca
tttctttcca tattagtagc tactactaaa tgattatctt ctttcatatc 420ttgtagataa
gatagactat ctttatcttt attagtagaa aatacttctg gccatacatc 480gttaaatttt
tttgttgttg ttagatataa tattaaatat ctagaggatc ctattatttg 540tggtaaaatg
tttatagagt aaaatgatct ggctattaaa cataggccag ttaccataga 600atgctgcttc
ccgttacagt gttttaccat aaccatagat ctgcctgtat tgttgataca 660tataacagct
gtaaatccta aaaaattcct atcataatta ttaatattag gtaattcatt 720tccatgtgaa
agatagacta attttatatc ctttacctcc aaataattat ttacatctct 780taaacaatct
attttaatat cattaactgg tattttataa tatccagaaa ggtttgaagg 840ggttgatgga
ataagtctat taacatcgtt aagtaaatta ttaatatcat gaatctttat 900tatattatac
ccataagtta aatttatatt tactttctca tcatctgact tagttagttt 960gtaataaggt
gtgtctgaaa aaattaaaag gtaattcgtt gaatgaagct gtatttgctg 1020tatcattttt
atctaatttt ggagatttag cagtacttac ttcattagaa gaagaatctg 1080ccagttcctg
tctattactg atatttcgtt tcattattat atgatttata ttttactttt 1140tcaattatat
atactcattt gactagttaa tcaataaaaa gaattcttct ttattctata 1200cttaaaaagt
gcaaataaat acaaaggttc ttg atg gga gct ggg caa tcc agc 1254
Met Gly Ala Gly Gln Ser Ser
1 5cca gca acc ggc tcg cag aac cag tct ggc
aac act ggc agc ata atc 1302Pro Ala Thr Gly Ser Gln Asn Gln Ser Gly
Asn Thr Gly Ser Ile Ile 10 15
20aac aac tac tac atg caa cag tac cag aac tcc atg gac aca cag ttg
1350Asn Asn Tyr Tyr Met Gln Gln Tyr Gln Asn Ser Met Asp Thr Gln Leu 25
30 35gga gac aat gcc atc agt gga ggc
tcc aac gag ggc tcc acg gac aca 1398Gly Asp Asn Ala Ile Ser Gly Gly
Ser Asn Glu Gly Ser Thr Asp Thr40 45 50
55act tca aca cac aca acc aac act caa aac aat gac tgg
ttc tcg aag 1446Thr Ser Thr His Thr Thr Asn Thr Gln Asn Asn Asp Trp
Phe Ser Lys 60 65 70ctc
gcc agt tca gct ttt acc ggt ctg ttc ggt gca ctg ctc gcc gac 1494Leu
Ala Ser Ser Ala Phe Thr Gly Leu Phe Gly Ala Leu Leu Ala Asp 75
80 85aag aag aca gag gaa acg aca ctt
ctt gag gac cgc atc ctc acc acc 1542Lys Lys Thr Glu Glu Thr Thr Leu
Leu Glu Asp Arg Ile Leu Thr Thr 90 95
100cgc aac ggg cac acc acc tcg acg acc caa tcg agt gtg ggt gtc aca
1590Arg Asn Gly His Thr Thr Ser Thr Thr Gln Ser Ser Val Gly Val Thr
105 110 115cac ggg tac tcc aca gag gag
gac cac gtt gct ggg ccc aac aca tcg 1638His Gly Tyr Ser Thr Glu Glu
Asp His Val Ala Gly Pro Asn Thr Ser120 125
130 135ggc ctg gag acg cga gtg gtg cag gca gag aga ttc
tac aaa aag tac 1686Gly Leu Glu Thr Arg Val Val Gln Ala Glu Arg Phe
Tyr Lys Lys Tyr 140 145
150ttg ttt gac tgg aca acg gac aag gca ttt gga cac ctg gaa aag ctg
1734Leu Phe Asp Trp Thr Thr Asp Lys Ala Phe Gly His Leu Glu Lys Leu
155 160 165gag ctc ccg tcc gac cac
cac ggt gtc ttt gga cac ttg gtg gac tcg 1782Glu Leu Pro Ser Asp His
His Gly Val Phe Gly His Leu Val Asp Ser 170 175
180tac gcc tat atg aga aat ggc tgg gat gtt gag gtg tcc gct
gtt ggc 1830Tyr Ala Tyr Met Arg Asn Gly Trp Asp Val Glu Val Ser Ala
Val Gly 185 190 195aac cag ttc aac ggc
ggg tgc ctc ctg gtg gcc atg gta cct gaa tgg 1878Asn Gln Phe Asn Gly
Gly Cys Leu Leu Val Ala Met Val Pro Glu Trp200 205
210 215aag gaa ttt gac aca cgg gag aaa tac caa
ctc acc ctt ttc ccg cac 1926Lys Glu Phe Asp Thr Arg Glu Lys Tyr Gln
Leu Thr Leu Phe Pro His 220 225
230cag ttt att agc ccc aga act aac atg act gcc cac atc acg gtc ccc
1974Gln Phe Ile Ser Pro Arg Thr Asn Met Thr Ala His Ile Thr Val Pro
235 240 245tac ctt ggt gtg aac agg
tat gat cag tac aag aag cat aag ccc tgg 2022Tyr Leu Gly Val Asn Arg
Tyr Asp Gln Tyr Lys Lys His Lys Pro Trp 250 255
260aca ttg gtt gtc atg gtc gtg tcg cca ctt acg gtc aac aac
act agt 2070Thr Leu Val Val Met Val Val Ser Pro Leu Thr Val Asn Asn
Thr Ser 265 270 275gcg gca caa atc aag
gtc tac gcc aac ata gct ccg acc tat gtt cac 2118Ala Ala Gln Ile Lys
Val Tyr Ala Asn Ile Ala Pro Thr Tyr Val His280 285
290 295gtg gcc ggt gaa ctc ccc tcg aaa gag ggg
att ttc ccg gtt gca tgt 2166Val Ala Gly Glu Leu Pro Ser Lys Glu Gly
Ile Phe Pro Val Ala Cys 300 305
310gcg gac ggt tac gga gga ttg gtg acg aca gac ccg aag aca gct gac
2214Ala Asp Gly Tyr Gly Gly Leu Val Thr Thr Asp Pro Lys Thr Ala Asp
315 320 325cct gct tat ggc aag gtg
tac aac ccg cct agg act aac tac cct ggg 2262Pro Ala Tyr Gly Lys Val
Tyr Asn Pro Pro Arg Thr Asn Tyr Pro Gly 330 335
340cgc ttc acc aac ctg ttg gac gtg gcc gaa gcg tgt ccc act
ttc ctc 2310Arg Phe Thr Asn Leu Leu Asp Val Ala Glu Ala Cys Pro Thr
Phe Leu 345 350 355tgc ttt gac gac ggg
aaa ccg tac gtc acc acg cgg acg gat gac acc 2358Cys Phe Asp Asp Gly
Lys Pro Tyr Val Thr Thr Arg Thr Asp Asp Thr360 365
370 375cga ctt ttg gcc aag ttt gac ctt tcc ctt
gcc gca aaa cat atg tcc 2406Arg Leu Leu Ala Lys Phe Asp Leu Ser Leu
Ala Ala Lys His Met Ser 380 385
390aac aca tac ctg tca ggg att gct cag tac tac aca cag tac tct ggc
2454Asn Thr Tyr Leu Ser Gly Ile Ala Gln Tyr Tyr Thr Gln Tyr Ser Gly
395 400 405acc atc aat ttg cat ttc
atg ttt aca ggt tcc act gat tca aag gcc 2502Thr Ile Asn Leu His Phe
Met Phe Thr Gly Ser Thr Asp Ser Lys Ala 410 415
420cga tac atg gtg gcc tac atc cca cct ggg gtg gag aca cca
ccg gac 2550Arg Tyr Met Val Ala Tyr Ile Pro Pro Gly Val Glu Thr Pro
Pro Asp 425 430 435aca cct gaa agg gct
gcc cac tgc att cac gct gaa tgg gac act gga 2598Thr Pro Glu Arg Ala
Ala His Cys Ile His Ala Glu Trp Asp Thr Gly440 445
450 455cta aac tcc aaa ttc act ttc tca atc ccg
tac gta tcc gcc gcg gat 2646Leu Asn Ser Lys Phe Thr Phe Ser Ile Pro
Tyr Val Ser Ala Ala Asp 460 465
470tac gcg tac aca gcg tct gac acg gca gaa aca atc aac gta cag gga
2694Tyr Ala Tyr Thr Ala Ser Asp Thr Ala Glu Thr Ile Asn Val Gln Gly
475 480 485tgg gtc tgc atc tac caa
att aca cac ggg aag gct gaa aat gac acc 2742Trp Val Cys Ile Tyr Gln
Ile Thr His Gly Lys Ala Glu Asn Asp Thr 490 495
500ttg gtc gtg tcg gtt agc gcc ggc aaa gac ttt gag ttg cgc
ctc ccg 2790Leu Val Val Ser Val Ser Ala Gly Lys Asp Phe Glu Leu Arg
Leu Pro 505 510 515att gac ccc cgc cag
cag acc acc gct acc ggg gaa tca gca gac ccg 2838Ile Asp Pro Arg Gln
Gln Thr Thr Ala Thr Gly Glu Ser Ala Asp Pro520 525
530 535gtc acc acc acc gtg gag aac tac ggc ggt
gag aca caa atc cag aga 2886Val Thr Thr Thr Val Glu Asn Tyr Gly Gly
Glu Thr Gln Ile Gln Arg 540 545
550cgt cac cac acg gac att ggt ttc atc atg gac aga ttt gtg aag atc
2934Arg His His Thr Asp Ile Gly Phe Ile Met Asp Arg Phe Val Lys Ile
555 560 565caa agc ttg agc cca aca
cat gtc att gac ctc atg cag gct cac caa 2982Gln Ser Leu Ser Pro Thr
His Val Ile Asp Leu Met Gln Ala His Gln 570 575
580cac ggt ctg gtg ggt gcc ttg ctg cgt gca gcc acg tac tac
ttt tct 3030His Gly Leu Val Gly Ala Leu Leu Arg Ala Ala Thr Tyr Tyr
Phe Ser 585 590 595gac ctg gaa att gtt
gta cgg cac gaa ggc aat ctg acc tgg gtg ccc 3078Asp Leu Glu Ile Val
Val Arg His Glu Gly Asn Leu Thr Trp Val Pro600 605
610 615aac ggc gcc cct gaa tca gcc ctg ttg aac
acc agc aac ccc act gcc 3126Asn Gly Ala Pro Glu Ser Ala Leu Leu Asn
Thr Ser Asn Pro Thr Ala 620 625
630tac aac aag gca cca ttc acg aga ctc gct ctc ccc tac act gcg ccg
3174Tyr Asn Lys Ala Pro Phe Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro
635 640 645cac cgt gtg ctg gca aca
gtg tac aac ggg acg agt aag tat gct gtg 3222His Arg Val Leu Ala Thr
Val Tyr Asn Gly Thr Ser Lys Tyr Ala Val 650 655
660ggt ggt tca ggc aga aga ggc gac atg ggg tct ctc gcg gcg
cga gtc 3270Gly Gly Ser Gly Arg Arg Gly Asp Met Gly Ser Leu Ala Ala
Arg Val 665 670 675gtg aaa cag ctt cct
gct tca ttt aac tac ggt gca atc aag gcc gac 3318Val Lys Gln Leu Pro
Ala Ser Phe Asn Tyr Gly Ala Ile Lys Ala Asp680 685
690 695gcc atc cac gaa ctt ctc gtg cgc atg aaa
cgg gcc gag ctc tac tgc 3366Ala Ile His Glu Leu Leu Val Arg Met Lys
Arg Ala Glu Leu Tyr Cys 700 705
710ccc aga ccg ctg ttg gca ata gag gtg tct tcg caa gac agg cac aag
3414Pro Arg Pro Leu Leu Ala Ile Glu Val Ser Ser Gln Asp Arg His Lys
715 720 725caa aag atc att gca cca
gca aag cag ctt ctg aat ttt gac ctg ctc 3462Gln Lys Ile Ile Ala Pro
Ala Lys Gln Leu Leu Asn Phe Asp Leu Leu 730 735
740aag ttg gcc gga gac gtt gag tcc aac ccc ggg cca ttc ttc
ttt gct 3510Lys Leu Ala Gly Asp Val Glu Ser Asn Pro Gly Pro Phe Phe
Phe Ala 745 750 755gac gtt agg tca aac
ttt tca aag ttg gta gac aca atc aac cag atg 3558Asp Val Arg Ser Asn
Phe Ser Lys Leu Val Asp Thr Ile Asn Gln Met760 765
770 775cag gag gac atg tcc aca aaa cac ggg ccc
gac ttc aac cgg ttg gtg 3606Gln Glu Asp Met Ser Thr Lys His Gly Pro
Asp Phe Asn Arg Leu Val 780 785
790tcc gca ttt gag gaa ttg gcc act gga gtt aaa gct atc agg acc ggt
3654Ser Ala Phe Glu Glu Leu Ala Thr Gly Val Lys Ala Ile Arg Thr Gly
795 800 805ctc gac gag gcc aaa ccc
tgg tac aag ctt atc aaa ctc cta agc cgc 3702Leu Asp Glu Ala Lys Pro
Trp Tyr Lys Leu Ile Lys Leu Leu Ser Arg 810 815
820ctg tcg tgc atg gcc gct gtg gca gca cgg tcc aag gac cca
gtc ctt 3750Leu Ser Cys Met Ala Ala Val Ala Ala Arg Ser Lys Asp Pro
Val Leu 825 830 835gtg gcc atc atg ctg
gcc gac acc ggt ctc gag cgt cag aga cct ctg 3798Val Ala Ile Met Leu
Ala Asp Thr Gly Leu Glu Arg Gln Arg Pro Leu840 845
850 855aaa gtg aga gct aag ctc cca cag cag gaa
gga cct tac gct ggc ccg 3846Lys Val Arg Ala Lys Leu Pro Gln Gln Glu
Gly Pro Tyr Ala Gly Pro 860 865
870ttg gag aga cag aaa ccg ctg aaa gtg aaa gca aaa gcc ccg gtc gtc
3894Leu Glu Arg Gln Lys Pro Leu Lys Val Lys Ala Lys Ala Pro Val Val
875 880 885aag gaa gga cct tac gag
gga ccg gtg aag aag cct gtc gct ttg aaa 3942Lys Glu Gly Pro Tyr Glu
Gly Pro Val Lys Lys Pro Val Ala Leu Lys 890 895
900gtg aaa gct aag aac ttg ata gtc act gag agt ggt gcc cca
ccg acc 3990Val Lys Ala Lys Asn Leu Ile Val Thr Glu Ser Gly Ala Pro
Pro Thr 905 910 915gac ttg caa aag atg
gtc atg ggc aac aca aag cct gtt gag ctc atc 4038Asp Leu Gln Lys Met
Val Met Gly Asn Thr Lys Pro Val Glu Leu Ile920 925
930 935ctt gac ggg aag aca gta gcc atc tgt tgt
gct act gga gtg ttt ggc 4086Leu Asp Gly Lys Thr Val Ala Ile Cys Cys
Ala Thr Gly Val Phe Gly 940 945
950act gct tac ctc gtg cct cgt cat ctt ttc gca gag aag tat gac aag
4134Thr Ala Tyr Leu Val Pro Arg His Leu Phe Ala Glu Lys Tyr Asp Lys
955 960 965atc atg ctg gat ggc aga
gcc atg aca gac agt gac tac aga gtg ttt 4182Ile Met Leu Asp Gly Arg
Ala Met Thr Asp Ser Asp Tyr Arg Val Phe 970 975
980gag ttt gag att aaa gta aaa gga cag gac atg ctc tca gac
gct gcg 4230Glu Phe Glu Ile Lys Val Lys Gly Gln Asp Met Leu Ser Asp
Ala Ala 985 990 995ctc atg gtg ctc cac
cgt ggg aac cgc gtg aga gat atc acg aaa cac 4278Leu Met Val Leu His
Arg Gly Asn Arg Val Arg Asp Ile Thr Lys His1000 1005
1010 1015ttt cgt gat aca gca aga atg aag aaa ggc
acc ccc gtc gtc ggt gtg 4326Phe Arg Asp Thr Ala Arg Met Lys Lys Gly
Thr Pro Val Val Gly Val 1020 1025
1030gtc aac aac gcc gac gtt ggg aga ctg att ttc tct ggt gag gcc ctc
4374Val Asn Asn Ala Asp Val Gly Arg Leu Ile Phe Ser Gly Glu Ala Leu
1035 1040 1045acc tac aag gat att
gta gtg tgc atg gac gga gac acc atg cct ggc 4422Thr Tyr Lys Asp Ile
Val Val Cys Met Asp Gly Asp Thr Met Pro Gly 1050
1055 1060ctc ttt gcc tac aaa gcc gcc acc aag gca ggc tac
tgt gga gga gcc 4470Leu Phe Ala Tyr Lys Ala Ala Thr Lys Ala Gly Tyr
Cys Gly Gly Ala 1065 1070 1075gtt ctc
gcc aag gac ggg gcc gac act ttc atc gtc ggc act cac tcc 4518Val Leu
Ala Lys Asp Gly Ala Asp Thr Phe Ile Val Gly Thr His Ser1080
1085 1090 1095gca gga ggc aat gga gtt gga
tac tgc tca tgc gtt tcc agg tcc atg 4566Ala Gly Gly Asn Gly Val Gly
Tyr Cys Ser Cys Val Ser Arg Ser Met 1100
1105 1110ctt ctc aga atg aag gca cac gtt gac cct gaa cca
caa cac gag 4611Leu Leu Arg Met Lys Ala His Val Asp Pro Glu Pro
Gln His Glu 1115 1120
1125tagtaatttt tctgcagccc gggtttttat agctaattag tcattttttc gtaagtaagt
4671atttttattt aatacttttt attgtactta tgttaaatat aactgatgat aacaaaatcc
4731attatgtatt atttataact gtaatttctt tagcgtagtt agatgtccaa tctctctcaa
4791atacatcggc tatcttttta gtgagatttt gatctatgca gttgaaactt atgaacgcgt
4851gatgattaaa atgtgaaccg tccaaatttg cagtcattat atgagcgtat ctattatcta
4911ctatcatcat ctttgagtta ttaatatcat ctactttaga attgatagga aatatgaata
4971cctttgtagt aatatctata ctatctacac ctaactcatt aagacttttg atag
5025271126PRTFoot-and-mouth disease virus 27Met Gly Ala Gly Gln Ser Ser
Pro Ala Thr Gly Ser Gln Asn Gln Ser1 5 10
15Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr Met Gln
Gln Tyr Gln 20 25 30Asn Ser
Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly Gly Ser 35
40 45Asn Glu Gly Ser Thr Asp Thr Thr Ser Thr
His Thr Thr Asn Thr Gln 50 55 60Asn
Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu65
70 75 80Phe Gly Ala Leu Leu Ala
Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu 85
90 95Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His Thr
Thr Ser Thr Thr 100 105 110Gln
Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu Asp His 115
120 125Val Ala Gly Pro Asn Thr Ser Gly Leu
Glu Thr Arg Val Val Gln Ala 130 135
140Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys Ala145
150 155 160Phe Gly His Leu
Glu Lys Leu Glu Leu Pro Ser Asp His His Gly Val 165
170 175Phe Gly His Leu Val Asp Ser Tyr Ala Tyr
Met Arg Asn Gly Trp Asp 180 185
190Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys Leu Leu
195 200 205Val Ala Met Val Pro Glu Trp
Lys Glu Phe Asp Thr Arg Glu Lys Tyr 210 215
220Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro Arg Thr Asn
Met225 230 235 240Thr Ala
His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln
245 250 255Tyr Lys Lys His Lys Pro Trp
Thr Leu Val Val Met Val Val Ser Pro 260 265
270Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys Val Tyr
Ala Asn 275 280 285Ile Ala Pro Thr
Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu 290
295 300Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr Gly
Gly Leu Val Thr305 310 315
320Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn Pro
325 330 335Pro Arg Thr Asn Tyr
Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala 340
345 350Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly
Lys Pro Tyr Val 355 360 365Thr Thr
Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser 370
375 380Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu
Ser Gly Ile Ala Gln385 390 395
400Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe Thr
405 410 415Gly Ser Thr Asp
Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro 420
425 430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg
Ala Ala His Cys Ile 435 440 445His
Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile 450
455 460Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr
Thr Ala Ser Asp Thr Ala465 470 475
480Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr Gln Ile Thr
His 485 490 495Gly Lys Ala
Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys 500
505 510Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro
Arg Gln Gln Thr Thr Ala 515 520
525Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu Asn Tyr Gly 530
535 540Gly Glu Thr Gln Ile Gln Arg Arg
His His Thr Asp Ile Gly Phe Ile545 550
555 560Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser Pro
Thr His Val Ile 565 570
575Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
580 585 590Ala Ala Thr Tyr Tyr Phe
Ser Asp Leu Glu Ile Val Val Arg His Glu 595 600
605Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ser Ala
Leu Leu 610 615 620Asn Thr Ser Asn Pro
Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu625 630
635 640Ala Leu Pro Tyr Thr Ala Pro His Arg Val
Leu Ala Thr Val Tyr Asn 645 650
655Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly Asp Met
660 665 670Gly Ser Leu Ala Ala
Arg Val Val Lys Gln Leu Pro Ala Ser Phe Asn 675
680 685Tyr Gly Ala Ile Lys Ala Asp Ala Ile His Glu Leu
Leu Val Arg Met 690 695 700Lys Arg Ala
Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val705
710 715 720Ser Ser Gln Asp Arg His Lys
Gln Lys Ile Ile Ala Pro Ala Lys Gln 725
730 735Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp
Val Glu Ser Asn 740 745 750Pro
Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser Lys Leu 755
760 765Val Asp Thr Ile Asn Gln Met Gln Glu
Asp Met Ser Thr Lys His Gly 770 775
780Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr Gly785
790 795 800Val Lys Ala Ile
Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr Lys 805
810 815Leu Ile Lys Leu Leu Ser Arg Leu Ser Cys
Met Ala Ala Val Ala Ala 820 825
830Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp Thr Gly
835 840 845Leu Glu Arg Gln Arg Pro Leu
Lys Val Arg Ala Lys Leu Pro Gln Gln 850 855
860Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys Pro Leu Lys
Val865 870 875 880Lys Ala
Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro Val
885 890 895Lys Lys Pro Val Ala Leu Lys
Val Lys Ala Lys Asn Leu Ile Val Thr 900 905
910Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met Val Met
Gly Asn 915 920 925Thr Lys Pro Val
Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys 930
935 940Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu Val
Pro Arg His Leu945 950 955
960Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met Thr
965 970 975Asp Ser Asp Tyr Arg
Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln 980
985 990Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu His
Arg Gly Asn Arg 995 1000 1005Val Arg
Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys 1010
1015 1020Gly Thr Pro Val Val Gly Val Val Asn Asn Ala
Asp Val Gly Arg Leu1025 1030 1035
1040Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys Met
1045 1050 1055Asp Gly Asp Thr
Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys 1060
1065 1070Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys
Asp Gly Ala Asp Thr 1075 1080
1085Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val Gly Tyr Cys
1090 1095 1100Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met Lys Ala His Val Asp1105 1110
1115 1120Pro Glu Pro Gln His Glu
11252827DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 28gaattctgag ataaagtgaa aatatat
272930DNAArtificial SequenceDescription of Artificial
Sequence Synthetic primer 29acaattattt aggtttaatc atgggagctg
303030DNAArtificial SequenceDescription of
Artificial Sequence Synthetic primer 30cagctcccat gattaaacct
aaataattgt 303129DNAArtificial
SequenceDescription of Artificial Sequence Synthetic primer
31aggtttaatc atgggagctg ggcaatcca
293223DNAArtificial SequenceDescription of Artificial Sequence Synthetic
primer 32tgcctcctgg tggccatggt acc
233323DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 33ggtaccatgg ccaccaggag gca
23346PRTArtificial SequenceDescription of Artificial
Sequence Synthetic peptide 34Met Gly Ala Gly Gln Ser1
5358PRTArtificial SequenceDescription of Artificial Sequence Synthetic
peptide 35Cys Leu Leu Val Ala Met Val Pro1
5365037DNAFoot-and-mouth disease virusCDS(1246)..(4623) 36ttcataaata
caagtttgat taaacttaag ttgttctaaa gttctttcct ccgaaggtat 60agaacaaagt
atttcttcta catccttact atttattgca gcttttaaca gcctatcacg 120tatcctattt
ttagtattgg tagaacgttt tagttctaaa gttaaaatat tagacataat 180tggcatattg
cttattcctt gcatagttga gtctgtagat cgtttcagta tatcactgat 240taatgtacta
ctgttatgat gaaatataga atcgatattg gcatttaact gttttgttat 300actaagtcta
gattttaaat cttctagtaa tatgctattt aatataaaag cttccacgtt 360tttgtataca
tttctttcca tattagtagc tactactaaa tgattatctt ctttcatatc 420ttgtagataa
gatagactat ctttatcttt attagtagaa aatacttctg gccatacatc 480gttaaatttt
tttgttgttg ttagatataa tattaaatat ctagaggatc ctattatttg 540tggtaaaatg
tttatagagt aaaatgatct ggctattaaa cataggccag ttaccataga 600atgctgcttc
ccgttacagt gttttaccat aaccatagat ctgcctgtat tgttgataca 660tataacagct
gtaaatccta aaaaattcct atcataatta ttaatattag gtaattcatt 720tccatgtgaa
agatagacta attttatatc ctttacctcc aaataattat ttacatctct 780taaacaatct
attttaatat cattaactgg tattttataa tatccagaaa ggtttgaagg 840ggttgatgga
ataagtctat taacatcgtt aagtaaatta ttaatatcat gaatctttat 900tatattatac
ccataagtta aatttatatt tactttctca tcatctgact tagttagttt 960gtaataaggt
gtgtctgaaa aaattaaaag gtaattcgtt gaatgaagct gtatttgctg 1020tatcattttt
atctaatttt ggagatttag cagtacttac ttcattagaa gaagaatctg 1080ccagttcctg
tctattactg atatttcgtt tcattattat atgatttata ttttactttt 1140tcaattatat
atactcattt gactagttaa tcaataaaaa gaattctgag ataaagtgaa 1200aatatatatc
attatattac aaagtacaat tatttaggtt taatc atg gga gct ggg 1257
Met Gly Ala Gly
1caa tcc agc cca gca acc ggc tcg cag aac cag
tct ggc aac act ggc 1305Gln Ser Ser Pro Ala Thr Gly Ser Gln Asn Gln
Ser Gly Asn Thr Gly5 10 15
20agc ata atc aac aac tac tac atg caa cag tac cag aac tcc atg gac
1353Ser Ile Ile Asn Asn Tyr Tyr Met Gln Gln Tyr Gln Asn Ser Met Asp
25 30 35aca cag ttg gga gac
aat gcc atc agt gga ggc tcc aac gag ggc tcc 1401Thr Gln Leu Gly Asp
Asn Ala Ile Ser Gly Gly Ser Asn Glu Gly Ser 40
45 50acg gac aca act tca aca cac aca acc aac act caa
aac aat gac tgg 1449Thr Asp Thr Thr Ser Thr His Thr Thr Asn Thr Gln
Asn Asn Asp Trp 55 60 65ttc tcg
aag ctc gcc agt tca gct ttt acc ggt ctg ttc ggt gca ctg 1497Phe Ser
Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu Phe Gly Ala Leu 70
75 80ctc gcc gac aag aag aca gag gaa acg aca ctt
ctt gag gac cgc atc 1545Leu Ala Asp Lys Lys Thr Glu Glu Thr Thr Leu
Leu Glu Asp Arg Ile85 90 95
100ctc acc acc cgc aac ggg cac acc acc tcg acg acc caa tcg agt gtg
1593Leu Thr Thr Arg Asn Gly His Thr Thr Ser Thr Thr Gln Ser Ser Val
105 110 115ggt gtc aca cac ggg
tac tcc aca gag gag gac cac gtt gct ggg ccc 1641Gly Val Thr His Gly
Tyr Ser Thr Glu Glu Asp His Val Ala Gly Pro 120
125 130aac aca tcg ggc ctg gag acg cga gtg gtg cag gca
gag aga ttc tac 1689Asn Thr Ser Gly Leu Glu Thr Arg Val Val Gln Ala
Glu Arg Phe Tyr 135 140 145aaa aag
tac ttg ttt gac tgg aca acg gac aag gca ttt gga cac ctg 1737Lys Lys
Tyr Leu Phe Asp Trp Thr Thr Asp Lys Ala Phe Gly His Leu 150
155 160gaa aag ctg gag ctc ccg tcc gac cac cac ggt
gtc ttt gga cac ttg 1785Glu Lys Leu Glu Leu Pro Ser Asp His His Gly
Val Phe Gly His Leu165 170 175
180gtg gac tcg tac gcc tat atg aga aat ggc tgg gat gtt gag gtg tcc
1833Val Asp Ser Tyr Ala Tyr Met Arg Asn Gly Trp Asp Val Glu Val Ser
185 190 195gct gtt ggc aac cag
ttc aac ggc ggg tgc ctc ctg gtg gcc atg gta 1881Ala Val Gly Asn Gln
Phe Asn Gly Gly Cys Leu Leu Val Ala Met Val 200
205 210cct gaa tgg aag gaa ttt gac aca cgg gag aaa tac
caa ctc acc ctt 1929Pro Glu Trp Lys Glu Phe Asp Thr Arg Glu Lys Tyr
Gln Leu Thr Leu 215 220 225ttc ccg
cac cag ttt att agc ccc aga act aac atg act gcc cac atc 1977Phe Pro
His Gln Phe Ile Ser Pro Arg Thr Asn Met Thr Ala His Ile 230
235 240acg gtc ccc tac ctt ggt gtg aac agg tat gat
cag tac aag aag cat 2025Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp
Gln Tyr Lys Lys His245 250 255
260aag ccc tgg aca ttg gtt gtc atg gtc gtg tcg cca ctt acg gtc aac
2073Lys Pro Trp Thr Leu Val Val Met Val Val Ser Pro Leu Thr Val Asn
265 270 275aac act agt gcg gca
caa atc aag gtc tac gcc aac ata gct ccg acc 2121Asn Thr Ser Ala Ala
Gln Ile Lys Val Tyr Ala Asn Ile Ala Pro Thr 280
285 290tat gtt cac gtg gcc ggt gaa ctc ccc tcg aaa gag
ggg att ttc ccg 2169Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu
Gly Ile Phe Pro 295 300 305gtt gca
tgt gcg gac ggt tac gga gga ttg gtg acg aca gac ccg aag 2217Val Ala
Cys Ala Asp Gly Tyr Gly Gly Leu Val Thr Thr Asp Pro Lys 310
315 320aca gct gac cct gct tat ggc aag gtg tac aac
ccg cct agg act aac 2265Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn
Pro Pro Arg Thr Asn325 330 335
340tac cct ggg cgc ttc acc aac ctg ttg gac gtg gcc gaa gcg tgt ccc
2313Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala Glu Ala Cys Pro
345 350 355act ttc ctc tgc ttt
gac gac ggg aaa ccg tac gtc acc acg cgg acg 2361Thr Phe Leu Cys Phe
Asp Asp Gly Lys Pro Tyr Val Thr Thr Arg Thr 360
365 370gat gac acc cga ctt ttg gcc aag ttt gac ctt tcc
ctt gcc gca aaa 2409Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser
Leu Ala Ala Lys 375 380 385cat atg
tcc aac aca tac ctg tca ggg att gct cag tac tac aca cag 2457His Met
Ser Asn Thr Tyr Leu Ser Gly Ile Ala Gln Tyr Tyr Thr Gln 390
395 400tac tct ggc acc atc aat ttg cat ttc atg ttt
aca ggt tcc act gat 2505Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe
Thr Gly Ser Thr Asp405 410 415
420tca aag gcc cga tac atg gtg gcc tac atc cca cct ggg gtg gag aca
2553Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro Gly Val Glu Thr
425 430 435cca ccg gac aca cct
gaa agg gct gcc cac tgc att cac gct gaa tgg 2601Pro Pro Asp Thr Pro
Glu Arg Ala Ala His Cys Ile His Ala Glu Trp 440
445 450gac act gga cta aac tcc aaa ttc act ttc tca atc
ccg tac gta tcc 2649Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile
Pro Tyr Val Ser 455 460 465gcc gcg
gat tac gcg tac aca gcg tct gac acg gca gaa aca atc aac 2697Ala Ala
Asp Tyr Ala Tyr Thr Ala Ser Asp Thr Ala Glu Thr Ile Asn 470
475 480gta cag gga tgg gtc tgc atc tac caa att aca
cac ggg aag gct gaa 2745Val Gln Gly Trp Val Cys Ile Tyr Gln Ile Thr
His Gly Lys Ala Glu485 490 495
500aat gac acc ttg gtc gtg tcg gtt agc gcc ggc aaa gac ttt gag ttg
2793Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys Asp Phe Glu Leu
505 510 515cgc ctc ccg att gac
ccc cgc cag cag acc acc gct acc ggg gaa tca 2841Arg Leu Pro Ile Asp
Pro Arg Gln Gln Thr Thr Ala Thr Gly Glu Ser 520
525 530gca gac ccg gtc acc acc acc gtg gag aac tac ggc
ggt gag aca caa 2889Ala Asp Pro Val Thr Thr Thr Val Glu Asn Tyr Gly
Gly Glu Thr Gln 535 540 545atc cag
aga cgt cac cac acg gac att ggt ttc atc atg gac aga ttt 2937Ile Gln
Arg Arg His His Thr Asp Ile Gly Phe Ile Met Asp Arg Phe 550
555 560gtg aag atc caa agc ttg agc cca aca cat gtc
att gac ctc atg cag 2985Val Lys Ile Gln Ser Leu Ser Pro Thr His Val
Ile Asp Leu Met Gln565 570 575
580gct cac caa cac ggt ctg gtg ggt gcc ttg ctg cgt gca gcc acg tac
3033Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg Ala Ala Thr Tyr
585 590 595tac ttt tct gac ctg
gaa att gtt gta cgg cac gaa ggc aat ctg acc 3081Tyr Phe Ser Asp Leu
Glu Ile Val Val Arg His Glu Gly Asn Leu Thr 600
605 610tgg gtg ccc aac ggc gcc cct gaa tca gcc ctg ttg
aac acc agc aac 3129Trp Val Pro Asn Gly Ala Pro Glu Ser Ala Leu Leu
Asn Thr Ser Asn 615 620 625ccc act
gcc tac aac aag gca cca ttc acg aga ctc gct ctc ccc tac 3177Pro Thr
Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu Ala Leu Pro Tyr 630
635 640act gcg ccg cac cgt gtg ctg gca aca gtg tac
aac ggg acg agt aag 3225Thr Ala Pro His Arg Val Leu Ala Thr Val Tyr
Asn Gly Thr Ser Lys645 650 655
660tat gct gtg ggt ggt tca ggc aga aga ggc gac atg ggg tct ctc gcg
3273Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly Asp Met Gly Ser Leu Ala
665 670 675gcg cga gtc gtg aaa
cag ctt cct gct tca ttt aac tac ggt gca atc 3321Ala Arg Val Val Lys
Gln Leu Pro Ala Ser Phe Asn Tyr Gly Ala Ile 680
685 690aag gcc gac gcc atc cac gaa ctt ctc gtg cgc atg
aaa cgg gcc gag 3369Lys Ala Asp Ala Ile His Glu Leu Leu Val Arg Met
Lys Arg Ala Glu 695 700 705ctc tac
tgc ccc aga ccg ctg ttg gca ata gag gtg tct tcg caa gac 3417Leu Tyr
Cys Pro Arg Pro Leu Leu Ala Ile Glu Val Ser Ser Gln Asp 710
715 720agg cac aag caa aag atc att gca cca gca aag
cag ctt ctg aat ttt 3465Arg His Lys Gln Lys Ile Ile Ala Pro Ala Lys
Gln Leu Leu Asn Phe725 730 735
740gac ctg ctc aag ttg gcc gga gac gtt gag tcc aac ccc ggg cca ttc
3513Asp Leu Leu Lys Leu Ala Gly Asp Val Glu Ser Asn Pro Gly Pro Phe
745 750 755ttc ttt gct gac gtt
agg tca aac ttt tca aag ttg gta gac aca atc 3561Phe Phe Ala Asp Val
Arg Ser Asn Phe Ser Lys Leu Val Asp Thr Ile 760
765 770aac cag atg cag gag gac atg tcc aca aaa cac ggg
ccc gac ttc aac 3609Asn Gln Met Gln Glu Asp Met Ser Thr Lys His Gly
Pro Asp Phe Asn 775 780 785cgg ttg
gtg tcc gca ttt gag gaa ttg gcc act gga gtt aaa gct atc 3657Arg Leu
Val Ser Ala Phe Glu Glu Leu Ala Thr Gly Val Lys Ala Ile 790
795 800agg acc ggt ctc gac gag gcc aaa ccc tgg tac
aag ctt atc aaa ctc 3705Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr
Lys Leu Ile Lys Leu805 810 815
820cta agc cgc ctg tcg tgc atg gcc gct gtg gca gca cgg tcc aag gac
3753Leu Ser Arg Leu Ser Cys Met Ala Ala Val Ala Ala Arg Ser Lys Asp
825 830 835cca gtc ctt gtg gcc
atc atg ctg gcc gac acc ggt ctc gag cgt cag 3801Pro Val Leu Val Ala
Ile Met Leu Ala Asp Thr Gly Leu Glu Arg Gln 840
845 850aga cct ctg aaa gtg aga gct aag ctc cca cag cag
gaa gga cct tac 3849Arg Pro Leu Lys Val Arg Ala Lys Leu Pro Gln Gln
Glu Gly Pro Tyr 855 860 865gct ggc
ccg ttg gag aga cag aaa ccg ctg aaa gtg aaa gca aaa gcc 3897Ala Gly
Pro Leu Glu Arg Gln Lys Pro Leu Lys Val Lys Ala Lys Ala 870
875 880ccg gtc gtc aag gaa gga cct tac gag gga ccg
gtg aag aag cct gtc 3945Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro
Val Lys Lys Pro Val885 890 895
900gct ttg aaa gtg aaa gct aag aac ttg ata gtc act gag agt ggt gcc
3993Ala Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr Glu Ser Gly Ala
905 910 915cca ccg acc gac ttg
caa aag atg gtc atg ggc aac aca aag cct gtt 4041Pro Pro Thr Asp Leu
Gln Lys Met Val Met Gly Asn Thr Lys Pro Val 920
925 930gag ctc atc ctt gac ggg aag aca gta gcc atc tgt
tgt gct act gga 4089Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys
Cys Ala Thr Gly 935 940 945gtg ttt
ggc act gct tac ctc gtg cct cgt cat ctt ttc gca gag aag 4137Val Phe
Gly Thr Ala Tyr Leu Val Pro Arg His Leu Phe Ala Glu Lys 950
955 960tat gac aag atc atg ctg gat ggc aga gcc atg
aca gac agt gac tac 4185Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met
Thr Asp Ser Asp Tyr965 970 975
980aga gtg ttt gag ttt gag att aaa gta aaa gga cag gac atg ctc tca
4233Arg Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln Asp Met Leu Ser
985 990 995gac gct gcg ctc atg
gtg ctc cac cgt ggg aac cgc gtg aga gat atc 4281Asp Ala Ala Leu Met
Val Leu His Arg Gly Asn Arg Val Arg Asp Ile 1000
1005 1010acg aaa cac ttt cgt gat aca gca aga atg aag aaa
ggc acc ccc gtc 4329Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys
Gly Thr Pro Val 1015 1020 1025gtc
ggt gtg gtc aac aac gcc gac gtt ggg aga ctg att ttc tct ggt 4377Val
Gly Val Val Asn Asn Ala Asp Val Gly Arg Leu Ile Phe Ser Gly 1030
1035 1040gag gcc ctc acc tac aag gat att gta gtg
tgc atg gac gga gac acc 4425Glu Ala Leu Thr Tyr Lys Asp Ile Val Val
Cys Met Asp Gly Asp Thr1045 1050 1055
1060atg cct ggc ctc ttt gcc tac aaa gcc gcc acc aag gca ggc tac
tgt 4473Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys Ala Gly Tyr
Cys 1065 1070 1075gga gga
gcc gtt ctc gcc aag gac ggg gcc gac act ttc atc gtc ggc 4521Gly Gly
Ala Val Leu Ala Lys Asp Gly Ala Asp Thr Phe Ile Val Gly 1080
1085 1090act cac tcc gca gga ggc aat gga gtt
gga tac tgc tca tgc gtt tcc 4569Thr His Ser Ala Gly Gly Asn Gly Val
Gly Tyr Cys Ser Cys Val Ser 1095 1100
1105agg tcc atg ctt ctc aga atg aag gca cac gtt gac cct gaa cca caa
4617Arg Ser Met Leu Leu Arg Met Lys Ala His Val Asp Pro Glu Pro Gln
1110 1115 1120cac gag tagtaatttt tctgcagccc
gggtttttat agctaattag tcattttttc 4673His Glu1125gtaagtaagt
atttttattt aatacttttt attgtactta tgttaaatat aactgatgat 4733aacaaaatcc
attatgtatt atttataact gtaatttctt tagcgtagtt agatgtccaa 4793tctctctcaa
atacatcggc tatcttttta gtgagatttt gatctatgca gttgaaactt 4853atgaacgcgt
gatgattaaa atgtgaaccg tccaaatttg cagtcattat atgagcgtat 4913ctattatcta
ctatcatcat ctttgagtta ttaatatcat ctactttaga attgatagga 4973aatatgaata
cctttgtagt aatatctata ctatctacac ctaactcatt aagacttttg 5033atag
5037371126PRTFoot-and-mouth disease virus 37Met Gly Ala Gly Gln Ser Ser
Pro Ala Thr Gly Ser Gln Asn Gln Ser1 5 10
15Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr Met Gln
Gln Tyr Gln 20 25 30Asn Ser
Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly Gly Ser 35
40 45Asn Glu Gly Ser Thr Asp Thr Thr Ser Thr
His Thr Thr Asn Thr Gln 50 55 60Asn
Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu65
70 75 80Phe Gly Ala Leu Leu Ala
Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu 85
90 95Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His Thr
Thr Ser Thr Thr 100 105 110Gln
Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu Asp His 115
120 125Val Ala Gly Pro Asn Thr Ser Gly Leu
Glu Thr Arg Val Val Gln Ala 130 135
140Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys Ala145
150 155 160Phe Gly His Leu
Glu Lys Leu Glu Leu Pro Ser Asp His His Gly Val 165
170 175Phe Gly His Leu Val Asp Ser Tyr Ala Tyr
Met Arg Asn Gly Trp Asp 180 185
190Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys Leu Leu
195 200 205Val Ala Met Val Pro Glu Trp
Lys Glu Phe Asp Thr Arg Glu Lys Tyr 210 215
220Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro Arg Thr Asn
Met225 230 235 240Thr Ala
His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln
245 250 255Tyr Lys Lys His Lys Pro Trp
Thr Leu Val Val Met Val Val Ser Pro 260 265
270Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys Val Tyr
Ala Asn 275 280 285Ile Ala Pro Thr
Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu 290
295 300Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr Gly
Gly Leu Val Thr305 310 315
320Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn Pro
325 330 335Pro Arg Thr Asn Tyr
Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala 340
345 350Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly
Lys Pro Tyr Val 355 360 365Thr Thr
Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser 370
375 380Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu
Ser Gly Ile Ala Gln385 390 395
400Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe Thr
405 410 415Gly Ser Thr Asp
Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro 420
425 430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg
Ala Ala His Cys Ile 435 440 445His
Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile 450
455 460Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr
Thr Ala Ser Asp Thr Ala465 470 475
480Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr Gln Ile Thr
His 485 490 495Gly Lys Ala
Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys 500
505 510Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro
Arg Gln Gln Thr Thr Ala 515 520
525Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu Asn Tyr Gly 530
535 540Gly Glu Thr Gln Ile Gln Arg Arg
His His Thr Asp Ile Gly Phe Ile545 550
555 560Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser Pro
Thr His Val Ile 565 570
575Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
580 585 590Ala Ala Thr Tyr Tyr Phe
Ser Asp Leu Glu Ile Val Val Arg His Glu 595 600
605Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ser Ala
Leu Leu 610 615 620Asn Thr Ser Asn Pro
Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu625 630
635 640Ala Leu Pro Tyr Thr Ala Pro His Arg Val
Leu Ala Thr Val Tyr Asn 645 650
655Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly Asp Met
660 665 670Gly Ser Leu Ala Ala
Arg Val Val Lys Gln Leu Pro Ala Ser Phe Asn 675
680 685Tyr Gly Ala Ile Lys Ala Asp Ala Ile His Glu Leu
Leu Val Arg Met 690 695 700Lys Arg Ala
Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val705
710 715 720Ser Ser Gln Asp Arg His Lys
Gln Lys Ile Ile Ala Pro Ala Lys Gln 725
730 735Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp
Val Glu Ser Asn 740 745 750Pro
Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser Lys Leu 755
760 765Val Asp Thr Ile Asn Gln Met Gln Glu
Asp Met Ser Thr Lys His Gly 770 775
780Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr Gly785
790 795 800Val Lys Ala Ile
Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr Lys 805
810 815Leu Ile Lys Leu Leu Ser Arg Leu Ser Cys
Met Ala Ala Val Ala Ala 820 825
830Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp Thr Gly
835 840 845Leu Glu Arg Gln Arg Pro Leu
Lys Val Arg Ala Lys Leu Pro Gln Gln 850 855
860Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys Pro Leu Lys
Val865 870 875 880Lys Ala
Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro Val
885 890 895Lys Lys Pro Val Ala Leu Lys
Val Lys Ala Lys Asn Leu Ile Val Thr 900 905
910Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met Val Met
Gly Asn 915 920 925Thr Lys Pro Val
Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys 930
935 940Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu Val
Pro Arg His Leu945 950 955
960Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met Thr
965 970 975Asp Ser Asp Tyr Arg
Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln 980
985 990Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu His
Arg Gly Asn Arg 995 1000 1005Val Arg
Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys 1010
1015 1020Gly Thr Pro Val Val Gly Val Val Asn Asn Ala
Asp Val Gly Arg Leu1025 1030 1035
1040Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys Met
1045 1050 1055Asp Gly Asp Thr
Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys 1060
1065 1070Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys
Asp Gly Ala Asp Thr 1075 1080
1085Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val Gly Tyr Cys
1090 1095 1100Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met Lys Ala His Val Asp1105 1110
1115 1120Pro Glu Pro Gln His Glu
11253824DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 38gaattctcaa aattgaaaat atat
243930DNAArtificial SequenceDescription of Artificial
Sequence Synthetic primer 39atatataatt acaatataaa atgggagctg
304030DNAArtificial SequenceDescription of
Artificial Sequence Synthetic primer 40cagctcccat tttatattgt
aattatatat 304128DNAArtificial
SequenceDescription of Artificial Sequence Synthetic primer
41caatataaaa tgggagctgg gcaatcca
284223DNAArtificial SequenceDescription of Artificial Sequence Synthetic
primer 42tgcctcctgg tggccatggt acc
234323DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 43ggtaccatgg ccaccaggag gca
23446PRTArtificial SequenceDescription of Artificial
Sequence Synthetic peptide 44Met Gly Ala Gly Gln Ser1
5458PRTArtificial SequenceDescription of Artificial Sequence Synthetic
peptide 45Cys Leu Leu Val Ala Met Val Pro1
5465010DNAFoot-and-mouth disease virusCDS(1219)..(4596) 46ttcataaata
caagtttgat taaacttaag ttgttctaaa gttctttcct ccgaaggtat 60agaacaaagt
atttcttcta catccttact atttattgca gcttttaaca gcctatcacg 120tatcctattt
ttagtattgg tagaacgttt tagttctaaa gttaaaatat tagacataat 180tggcatattg
cttattcctt gcatagttga gtctgtagat cgtttcagta tatcactgat 240taatgtacta
ctgttatgat gaaatataga atcgatattg gcatttaact gttttgttat 300actaagtcta
gattttaaat cttctagtaa tatgctattt aatataaaag cttccacgtt 360tttgtataca
tttctttcca tattagtagc tactactaaa tgattatctt ctttcatatc 420ttgtagataa
gatagactat ctttatcttt attagtagaa aatacttctg gccatacatc 480gttaaatttt
tttgttgttg ttagatataa tattaaatat ctagaggatc ctattatttg 540tggtaaaatg
tttatagagt aaaatgatct ggctattaaa cataggccag ttaccataga 600atgctgcttc
ccgttacagt gttttaccat aaccatagat ctgcctgtat tgttgataca 660tataacagct
gtaaatccta aaaaattcct atcataatta ttaatattag gtaattcatt 720tccatgtgaa
agatagacta attttatatc ctttacctcc aaataattat ttacatctct 780taaacaatct
attttaatat cattaactgg tattttataa tatccagaaa ggtttgaagg 840ggttgatgga
ataagtctat taacatcgtt aagtaaatta ttaatatcat gaatctttat 900tatattatac
ccataagtta aatttatatt tactttctca tcatctgact tagttagttt 960gtaataaggt
gtgtctgaaa aaattaaaag gtaattcgtt gaatgaagct gtatttgctg 1020tatcattttt
atctaatttt ggagatttag cagtacttac ttcattagaa gaagaatctg 1080ccagttcctg
tctattactg atatttcgtt tcattattat atgatttata ttttactttt 1140tcaattatat
atactcattt gactagttaa tcaataaaaa gaattctcaa aattgaaaat 1200atataattac
aatataaa atg gga gct ggg caa tcc agc cca gca acc ggc 1251
Met Gly Ala Gly Gln Ser Ser Pro Ala Thr Gly 1
5 10tcg cag aac cag tct ggc aac act ggc agc
ata atc aac aac tac tac 1299Ser Gln Asn Gln Ser Gly Asn Thr Gly Ser
Ile Ile Asn Asn Tyr Tyr 15 20
25atg caa cag tac cag aac tcc atg gac aca cag ttg gga gac aat gcc
1347Met Gln Gln Tyr Gln Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala
30 35 40atc agt gga ggc tcc aac gag
ggc tcc acg gac aca act tca aca cac 1395Ile Ser Gly Gly Ser Asn Glu
Gly Ser Thr Asp Thr Thr Ser Thr His 45 50
55aca acc aac act caa aac aat gac tgg ttc tcg aag ctc gcc agt tca
1443Thr Thr Asn Thr Gln Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser60
65 70 75gct ttt acc ggt
ctg ttc ggt gca ctg ctc gcc gac aag aag aca gag 1491Ala Phe Thr Gly
Leu Phe Gly Ala Leu Leu Ala Asp Lys Lys Thr Glu 80
85 90gaa acg aca ctt ctt gag gac cgc atc ctc
acc acc cgc aac ggg cac 1539Glu Thr Thr Leu Leu Glu Asp Arg Ile Leu
Thr Thr Arg Asn Gly His 95 100
105acc acc tcg acg acc caa tcg agt gtg ggt gtc aca cac ggg tac tcc
1587Thr Thr Ser Thr Thr Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser
110 115 120aca gag gag gac cac gtt gct
ggg ccc aac aca tcg ggc ctg gag acg 1635Thr Glu Glu Asp His Val Ala
Gly Pro Asn Thr Ser Gly Leu Glu Thr 125 130
135cga gtg gtg cag gca gag aga ttc tac aaa aag tac ttg ttt gac tgg
1683Arg Val Val Gln Ala Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp140
145 150 155aca acg gac aag
gca ttt gga cac ctg gaa aag ctg gag ctc ccg tcc 1731Thr Thr Asp Lys
Ala Phe Gly His Leu Glu Lys Leu Glu Leu Pro Ser 160
165 170gac cac cac ggt gtc ttt gga cac ttg gtg
gac tcg tac gcc tat atg 1779Asp His His Gly Val Phe Gly His Leu Val
Asp Ser Tyr Ala Tyr Met 175 180
185aga aat ggc tgg gat gtt gag gtg tcc gct gtt ggc aac cag ttc aac
1827Arg Asn Gly Trp Asp Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn
190 195 200ggc ggg tgc ctc ctg gtg gcc
atg gta cct gaa tgg aag gaa ttt gac 1875Gly Gly Cys Leu Leu Val Ala
Met Val Pro Glu Trp Lys Glu Phe Asp 205 210
215aca cgg gag aaa tac caa ctc acc ctt ttc ccg cac cag ttt att agc
1923Thr Arg Glu Lys Tyr Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser220
225 230 235ccc aga act aac
atg act gcc cac atc acg gtc ccc tac ctt ggt gtg 1971Pro Arg Thr Asn
Met Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val 240
245 250aac agg tat gat cag tac aag aag cat aag
ccc tgg aca ttg gtt gtc 2019Asn Arg Tyr Asp Gln Tyr Lys Lys His Lys
Pro Trp Thr Leu Val Val 255 260
265atg gtc gtg tcg cca ctt acg gtc aac aac act agt gcg gca caa atc
2067Met Val Val Ser Pro Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile
270 275 280aag gtc tac gcc aac ata gct
ccg acc tat gtt cac gtg gcc ggt gaa 2115Lys Val Tyr Ala Asn Ile Ala
Pro Thr Tyr Val His Val Ala Gly Glu 285 290
295ctc ccc tcg aaa gag ggg att ttc ccg gtt gca tgt gcg gac ggt tac
2163Leu Pro Ser Lys Glu Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr300
305 310 315gga gga ttg gtg
acg aca gac ccg aag aca gct gac cct gct tat ggc 2211Gly Gly Leu Val
Thr Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly 320
325 330aag gtg tac aac ccg cct agg act aac tac
cct ggg cgc ttc acc aac 2259Lys Val Tyr Asn Pro Pro Arg Thr Asn Tyr
Pro Gly Arg Phe Thr Asn 335 340
345ctg ttg gac gtg gcc gaa gcg tgt ccc act ttc ctc tgc ttt gac gac
2307Leu Leu Asp Val Ala Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp
350 355 360ggg aaa ccg tac gtc acc acg
cgg acg gat gac acc cga ctt ttg gcc 2355Gly Lys Pro Tyr Val Thr Thr
Arg Thr Asp Asp Thr Arg Leu Leu Ala 365 370
375aag ttt gac ctt tcc ctt gcc gca aaa cat atg tcc aac aca tac ctg
2403Lys Phe Asp Leu Ser Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu380
385 390 395tca ggg att gct
cag tac tac aca cag tac tct ggc acc atc aat ttg 2451Ser Gly Ile Ala
Gln Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu 400
405 410cat ttc atg ttt aca ggt tcc act gat tca
aag gcc cga tac atg gtg 2499His Phe Met Phe Thr Gly Ser Thr Asp Ser
Lys Ala Arg Tyr Met Val 415 420
425gcc tac atc cca cct ggg gtg gag aca cca ccg gac aca cct gaa agg
2547Ala Tyr Ile Pro Pro Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg
430 435 440gct gcc cac tgc att cac gct
gaa tgg gac act gga cta aac tcc aaa 2595Ala Ala His Cys Ile His Ala
Glu Trp Asp Thr Gly Leu Asn Ser Lys 445 450
455ttc act ttc tca atc ccg tac gta tcc gcc gcg gat tac gcg tac aca
2643Phe Thr Phe Ser Ile Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr Thr460
465 470 475gcg tct gac acg
gca gaa aca atc aac gta cag gga tgg gtc tgc atc 2691Ala Ser Asp Thr
Ala Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile 480
485 490tac caa att aca cac ggg aag gct gaa aat
gac acc ttg gtc gtg tcg 2739Tyr Gln Ile Thr His Gly Lys Ala Glu Asn
Asp Thr Leu Val Val Ser 495 500
505gtt agc gcc ggc aaa gac ttt gag ttg cgc ctc ccg att gac ccc cgc
2787Val Ser Ala Gly Lys Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg
510 515 520cag cag acc acc gct acc ggg
gaa tca gca gac ccg gtc acc acc acc 2835Gln Gln Thr Thr Ala Thr Gly
Glu Ser Ala Asp Pro Val Thr Thr Thr 525 530
535gtg gag aac tac ggc ggt gag aca caa atc cag aga cgt cac cac acg
2883Val Glu Asn Tyr Gly Gly Glu Thr Gln Ile Gln Arg Arg His His Thr540
545 550 555gac att ggt ttc
atc atg gac aga ttt gtg aag atc caa agc ttg agc 2931Asp Ile Gly Phe
Ile Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser 560
565 570cca aca cat gtc att gac ctc atg cag gct
cac caa cac ggt ctg gtg 2979Pro Thr His Val Ile Asp Leu Met Gln Ala
His Gln His Gly Leu Val 575 580
585ggt gcc ttg ctg cgt gca gcc acg tac tac ttt tct gac ctg gaa att
3027Gly Ala Leu Leu Arg Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile
590 595 600gtt gta cgg cac gaa ggc aat
ctg acc tgg gtg ccc aac ggc gcc cct 3075Val Val Arg His Glu Gly Asn
Leu Thr Trp Val Pro Asn Gly Ala Pro 605 610
615gaa tca gcc ctg ttg aac acc agc aac ccc act gcc tac aac aag gca
3123Glu Ser Ala Leu Leu Asn Thr Ser Asn Pro Thr Ala Tyr Asn Lys Ala620
625 630 635cca ttc acg aga
ctc gct ctc ccc tac act gcg ccg cac cgt gtg ctg 3171Pro Phe Thr Arg
Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu 640
645 650gca aca gtg tac aac ggg acg agt aag tat
gct gtg ggt ggt tca ggc 3219Ala Thr Val Tyr Asn Gly Thr Ser Lys Tyr
Ala Val Gly Gly Ser Gly 655 660
665aga aga ggc gac atg ggg tct ctc gcg gcg cga gtc gtg aaa cag ctt
3267Arg Arg Gly Asp Met Gly Ser Leu Ala Ala Arg Val Val Lys Gln Leu
670 675 680cct gct tca ttt aac tac ggt
gca atc aag gcc gac gcc atc cac gaa 3315Pro Ala Ser Phe Asn Tyr Gly
Ala Ile Lys Ala Asp Ala Ile His Glu 685 690
695ctt ctc gtg cgc atg aaa cgg gcc gag ctc tac tgc ccc aga ccg ctg
3363Leu Leu Val Arg Met Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu700
705 710 715ttg gca ata gag
gtg tct tcg caa gac agg cac aag caa aag atc att 3411Leu Ala Ile Glu
Val Ser Ser Gln Asp Arg His Lys Gln Lys Ile Ile 720
725 730gca cca gca aag cag ctt ctg aat ttt gac
ctg ctc aag ttg gcc gga 3459Ala Pro Ala Lys Gln Leu Leu Asn Phe Asp
Leu Leu Lys Leu Ala Gly 735 740
745gac gtt gag tcc aac ccc ggg cca ttc ttc ttt gct gac gtt agg tca
3507Asp Val Glu Ser Asn Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser
750 755 760aac ttt tca aag ttg gta gac
aca atc aac cag atg cag gag gac atg 3555Asn Phe Ser Lys Leu Val Asp
Thr Ile Asn Gln Met Gln Glu Asp Met 765 770
775tcc aca aaa cac ggg ccc gac ttc aac cgg ttg gtg tcc gca ttt gag
3603Ser Thr Lys His Gly Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu780
785 790 795gaa ttg gcc act
gga gtt aaa gct atc agg acc ggt ctc gac gag gcc 3651Glu Leu Ala Thr
Gly Val Lys Ala Ile Arg Thr Gly Leu Asp Glu Ala 800
805 810aaa ccc tgg tac aag ctt atc aaa ctc cta
agc cgc ctg tcg tgc atg 3699Lys Pro Trp Tyr Lys Leu Ile Lys Leu Leu
Ser Arg Leu Ser Cys Met 815 820
825gcc gct gtg gca gca cgg tcc aag gac cca gtc ctt gtg gcc atc atg
3747Ala Ala Val Ala Ala Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met
830 835 840ctg gcc gac acc ggt ctc gag
cgt cag aga cct ctg aaa gtg aga gct 3795Leu Ala Asp Thr Gly Leu Glu
Arg Gln Arg Pro Leu Lys Val Arg Ala 845 850
855aag ctc cca cag cag gaa gga cct tac gct ggc ccg ttg gag aga cag
3843Lys Leu Pro Gln Gln Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln860
865 870 875aaa ccg ctg aaa
gtg aaa gca aaa gcc ccg gtc gtc aag gaa gga cct 3891Lys Pro Leu Lys
Val Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro 880
885 890tac gag gga ccg gtg aag aag cct gtc gct
ttg aaa gtg aaa gct aag 3939Tyr Glu Gly Pro Val Lys Lys Pro Val Ala
Leu Lys Val Lys Ala Lys 895 900
905aac ttg ata gtc act gag agt ggt gcc cca ccg acc gac ttg caa aag
3987Asn Leu Ile Val Thr Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys
910 915 920atg gtc atg ggc aac aca aag
cct gtt gag ctc atc ctt gac ggg aag 4035Met Val Met Gly Asn Thr Lys
Pro Val Glu Leu Ile Leu Asp Gly Lys 925 930
935aca gta gcc atc tgt tgt gct act gga gtg ttt ggc act gct tac ctc
4083Thr Val Ala Ile Cys Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu940
945 950 955gtg cct cgt cat
ctt ttc gca gag aag tat gac aag atc atg ctg gat 4131Val Pro Arg His
Leu Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp 960
965 970ggc aga gcc atg aca gac agt gac tac aga
gtg ttt gag ttt gag att 4179Gly Arg Ala Met Thr Asp Ser Asp Tyr Arg
Val Phe Glu Phe Glu Ile 975 980
985aaa gta aaa gga cag gac atg ctc tca gac gct gcg ctc atg gtg ctc
4227Lys Val Lys Gly Gln Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu
990 995 1000cac cgt ggg aac cgc gtg aga
gat atc acg aaa cac ttt cgt gat aca 4275His Arg Gly Asn Arg Val Arg
Asp Ile Thr Lys His Phe Arg Asp Thr 1005 1010
1015gca aga atg aag aaa ggc acc ccc gtc gtc ggt gtg gtc aac aac gcc
4323Ala Arg Met Lys Lys Gly Thr Pro Val Val Gly Val Val Asn Asn
Ala1020 1025 1030 1035gac gtt
ggg aga ctg att ttc tct ggt gag gcc ctc acc tac aag gat 4371Asp Val
Gly Arg Leu Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp
1040 1045 1050att gta gtg tgc atg gac gga
gac acc atg cct ggc ctc ttt gcc tac 4419Ile Val Val Cys Met Asp Gly
Asp Thr Met Pro Gly Leu Phe Ala Tyr 1055 1060
1065aaa gcc gcc acc aag gca ggc tac tgt gga gga gcc gtt ctc
gcc aag 4467Lys Ala Ala Thr Lys Ala Gly Tyr Cys Gly Gly Ala Val Leu
Ala Lys 1070 1075 1080gac ggg gcc
gac act ttc atc gtc ggc act cac tcc gca gga ggc aat 4515Asp Gly Ala
Asp Thr Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn 1085
1090 1095gga gtt gga tac tgc tca tgc gtt tcc agg tcc atg
ctt ctc aga atg 4563Gly Val Gly Tyr Cys Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met1100 1105 1110
1115aag gca cac gtt gac cct gaa cca caa cac gag tagtaatttt tctgcagccc
4616Lys Ala His Val Asp Pro Glu Pro Gln His Glu 1120
1125gggtttttat agctaattag tcattttttc gtaagtaagt atttttattt
aatacttttt 4676attgtactta tgttaaatat aactgatgat aacaaaatcc attatgtatt
atttataact 4736gtaatttctt tagcgtagtt agatgtccaa tctctctcaa atacatcggc
tatcttttta 4796gtgagatttt gatctatgca gttgaaactt atgaacgcgt gatgattaaa
atgtgaaccg 4856tccaaatttg cagtcattat atgagcgtat ctattatcta ctatcatcat
ctttgagtta 4916ttaatatcat ctactttaga attgatagga aatatgaata cctttgtagt
aatatctata 4976ctatctacac ctaactcatt aagacttttg atag
5010471126PRTFoot-and-mouth disease virus 47Met Gly Ala Gly
Gln Ser Ser Pro Ala Thr Gly Ser Gln Asn Gln Ser1 5
10 15Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr
Tyr Met Gln Gln Tyr Gln 20 25
30Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly Gly Ser
35 40 45Asn Glu Gly Ser Thr Asp Thr Thr
Ser Thr His Thr Thr Asn Thr Gln 50 55
60Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu65
70 75 80Phe Gly Ala Leu Leu
Ala Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu 85
90 95Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His
Thr Thr Ser Thr Thr 100 105
110Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu Asp His
115 120 125Val Ala Gly Pro Asn Thr Ser
Gly Leu Glu Thr Arg Val Val Gln Ala 130 135
140Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys
Ala145 150 155 160Phe Gly
His Leu Glu Lys Leu Glu Leu Pro Ser Asp His His Gly Val
165 170 175Phe Gly His Leu Val Asp Ser
Tyr Ala Tyr Met Arg Asn Gly Trp Asp 180 185
190Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys
Leu Leu 195 200 205Val Ala Met Val
Pro Glu Trp Lys Glu Phe Asp Thr Arg Glu Lys Tyr 210
215 220Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro
Arg Thr Asn Met225 230 235
240Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln
245 250 255Tyr Lys Lys His Lys
Pro Trp Thr Leu Val Val Met Val Val Ser Pro 260
265 270Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys
Val Tyr Ala Asn 275 280 285Ile Ala
Pro Thr Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu 290
295 300Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr
Gly Gly Leu Val Thr305 310 315
320Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn Pro
325 330 335Pro Arg Thr Asn
Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala 340
345 350Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp
Gly Lys Pro Tyr Val 355 360 365Thr
Thr Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser 370
375 380Leu Ala Ala Lys His Met Ser Asn Thr Tyr
Leu Ser Gly Ile Ala Gln385 390 395
400Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe
Thr 405 410 415Gly Ser Thr
Asp Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro 420
425 430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu
Arg Ala Ala His Cys Ile 435 440
445His Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile 450
455 460Pro Tyr Val Ser Ala Ala Asp Tyr
Ala Tyr Thr Ala Ser Asp Thr Ala465 470
475 480Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr
Gln Ile Thr His 485 490
495Gly Lys Ala Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys
500 505 510Asp Phe Glu Leu Arg Leu
Pro Ile Asp Pro Arg Gln Gln Thr Thr Ala 515 520
525Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu Asn
Tyr Gly 530 535 540Gly Glu Thr Gln Ile
Gln Arg Arg His His Thr Asp Ile Gly Phe Ile545 550
555 560Met Asp Arg Phe Val Lys Ile Gln Ser Leu
Ser Pro Thr His Val Ile 565 570
575Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
580 585 590Ala Ala Thr Tyr Tyr
Phe Ser Asp Leu Glu Ile Val Val Arg His Glu 595
600 605Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu
Ser Ala Leu Leu 610 615 620Asn Thr Ser
Asn Pro Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu625
630 635 640Ala Leu Pro Tyr Thr Ala Pro
His Arg Val Leu Ala Thr Val Tyr Asn 645
650 655Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg
Arg Gly Asp Met 660 665 670Gly
Ser Leu Ala Ala Arg Val Val Lys Gln Leu Pro Ala Ser Phe Asn 675
680 685Tyr Gly Ala Ile Lys Ala Asp Ala Ile
His Glu Leu Leu Val Arg Met 690 695
700Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val705
710 715 720Ser Ser Gln Asp
Arg His Lys Gln Lys Ile Ile Ala Pro Ala Lys Gln 725
730 735Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala
Gly Asp Val Glu Ser Asn 740 745
750Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser Lys Leu
755 760 765Val Asp Thr Ile Asn Gln Met
Gln Glu Asp Met Ser Thr Lys His Gly 770 775
780Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr
Gly785 790 795 800Val Lys
Ala Ile Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr Lys
805 810 815Leu Ile Lys Leu Leu Ser Arg
Leu Ser Cys Met Ala Ala Val Ala Ala 820 825
830Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp
Thr Gly 835 840 845Leu Glu Arg Gln
Arg Pro Leu Lys Val Arg Ala Lys Leu Pro Gln Gln 850
855 860Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys
Pro Leu Lys Val865 870 875
880Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro Val
885 890 895Lys Lys Pro Val Ala
Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr 900
905 910Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met
Val Met Gly Asn 915 920 925Thr Lys
Pro Val Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys 930
935 940Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu
Val Pro Arg His Leu945 950 955
960Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met Thr
965 970 975Asp Ser Asp Tyr
Arg Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln 980
985 990Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu
His Arg Gly Asn Arg 995 1000 1005Val
Arg Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys 1010
1015 1020Gly Thr Pro Val Val Gly Val Val Asn Asn
Ala Asp Val Gly Arg Leu1025 1030 1035
1040Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys
Met 1045 1050 1055Asp Gly
Asp Thr Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys 1060
1065 1070Ala Gly Tyr Cys Gly Gly Ala Val Leu
Ala Lys Asp Gly Ala Asp Thr 1075 1080
1085Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val Gly Tyr Cys
1090 1095 1100Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met Lys Ala His Val Asp1105 1110
1115 1120Pro Glu Pro Gln His Glu
11254864DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 48gaattcaaaa gtagaaaata tattctaatt tattgcacgg
atgggagctg ggcaatccag 60ccca
644922DNAArtificial SequenceDescription of
Artificial Sequence Synthetic primer 49cttgccgcaa aacatatgtc ca
225022DNAArtificial
SequenceDescription of Artificial Sequence Synthetic primer
50tggacatatg ttttgcggca ag
225135DNAArtificial SequenceDescription of Artificial Sequence Synthetic
primer 51ctaatttatt gcacggatgg gagctgggca atcca
355237DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 52ctggattgcc cagctcccat ccgtgcaata aattaga
375328DNAArtificial SequenceDescription of Artificial
Sequence Synthetic primer 53gaaaatatat tctaatttat tgcacgga
285428DNAArtificial SequenceDescription of
Artificial Sequence Synthetic primer 54tccgtgcaat aaattagaat
atattttc 28558PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 55Met
Gly Ala Gly Gln Ser Ser Pro1 5567PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 56Leu
Ala Ala Lys His Met Ser1 5577PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 57Met
Gly Ala Gly Gln Ser Ser1 5585012DNAFoot-and-mouth disease
virusCDS(1221)..(4598) 58ttcataaata caagtttgat taaacttaag ttgttctaaa
gttctttcct ccgaaggtat 60agaacaaagt atttcttcta catccttact atttattgca
gcttttaaca gcctatcacg 120tatcctattt ttagtattgg tagaacgttt tagttctaaa
gttaaaatat tagacataat 180tggcatattg cttattcctt gcatagttga gtctgtagat
cgtttcagta tatcactgat 240taatgtacta ctgttatgat gaaatataga atcgatattg
gcatttaact gttttgttat 300actaagtcta gattttaaat cttctagtaa tatgctattt
aatataaaag cttccacgtt 360tttgtataca tttctttcca tattagtagc tactactaaa
tgattatctt ctttcatatc 420ttgtagataa gatagactat ctttatcttt attagtagaa
aatacttctg gccatacatc 480gttaaatttt tttgttgttg ttagatataa tattaaatat
ctagaggatc ctattatttg 540tggtaaaatg tttatagagt aaaatgatct ggctattaaa
cataggccag ttaccataga 600atgctgcttc ccgttacagt gttttaccat aaccatagat
ctgcctgtat tgttgataca 660tataacagct gtaaatccta aaaaattcct atcataatta
ttaatattag gtaattcatt 720tccatgtgaa agatagacta attttatatc ctttacctcc
aaataattat ttacatctct 780taaacaatct attttaatat cattaactgg tattttataa
tatccagaaa ggtttgaagg 840ggttgatgga ataagtctat taacatcgtt aagtaaatta
ttaatatcat gaatctttat 900tatattatac ccataagtta aatttatatt tactttctca
tcatctgact tagttagttt 960gtaataaggt gtgtctgaaa aaattaaaag gtaattcgtt
gaatgaagct gtatttgctg 1020tatcattttt atctaatttt ggagatttag cagtacttac
ttcattagaa gaagaatctg 1080ccagttcctg tctattactg atatttcgtt tcattattat
atgatttata ttttactttt 1140tcaattatat atactcattt gactagttaa tcaataaaaa
gaattcaaaa gtagaaaata 1200tattctaatt tattgcacgg atg gga gct ggg caa tcc
agc cca gca acc ggc 1253 Met Gly Ala Gly Gln Ser
Ser Pro Ala Thr Gly 1 5
10tcg cag aac cag tct ggc aac act ggc agc ata atc aac aac tac tac
1301Ser Gln Asn Gln Ser Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr
15 20 25atg caa cag tac cag aac
tcc atg gac aca cag ttg gga gac aat gcc 1349Met Gln Gln Tyr Gln Asn
Ser Met Asp Thr Gln Leu Gly Asp Asn Ala 30 35
40atc agt gga ggc tcc aac gag ggc tcc acg gac aca act tca
aca cac 1397Ile Ser Gly Gly Ser Asn Glu Gly Ser Thr Asp Thr Thr Ser
Thr His 45 50 55aca acc aac act caa
aac aat gac tgg ttc tcg aag ctc gcc agt tca 1445Thr Thr Asn Thr Gln
Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser60 65
70 75gct ttt acc ggt ctg ttc ggt gca ctg ctc
gcc gac aag aag aca gag 1493Ala Phe Thr Gly Leu Phe Gly Ala Leu Leu
Ala Asp Lys Lys Thr Glu 80 85
90gaa acg aca ctt ctt gag gac cgc atc ctc acc acc cgc aac ggg cac
1541Glu Thr Thr Leu Leu Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His
95 100 105acc acc tcg acg acc caa
tcg agt gtg ggt gtc aca cac ggg tac tcc 1589Thr Thr Ser Thr Thr Gln
Ser Ser Val Gly Val Thr His Gly Tyr Ser 110 115
120aca gag gag gac cac gtt gct ggg ccc aac aca tcg ggc ctg
gag acg 1637Thr Glu Glu Asp His Val Ala Gly Pro Asn Thr Ser Gly Leu
Glu Thr 125 130 135cga gtg gtg cag gca
gag aga ttc tac aaa aag tac ttg ttt gac tgg 1685Arg Val Val Gln Ala
Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp140 145
150 155aca acg gac aag gca ttt gga cac ctg gaa
aag ctg gag ctc ccg tcc 1733Thr Thr Asp Lys Ala Phe Gly His Leu Glu
Lys Leu Glu Leu Pro Ser 160 165
170gac cac cac ggt gtc ttt gga cac ttg gtg gac tcg tac gcc tat atg
1781Asp His His Gly Val Phe Gly His Leu Val Asp Ser Tyr Ala Tyr Met
175 180 185aga aat ggc tgg gat gtt
gag gtg tcc gct gtt ggc aac cag ttc aac 1829Arg Asn Gly Trp Asp Val
Glu Val Ser Ala Val Gly Asn Gln Phe Asn 190 195
200ggc ggg tgc ctc ctg gtg gcc atg gta cct gaa tgg aag gaa
ttt gac 1877Gly Gly Cys Leu Leu Val Ala Met Val Pro Glu Trp Lys Glu
Phe Asp 205 210 215aca cgg gag aaa tac
caa ctc acc ctt ttc ccg cac cag ttt att agc 1925Thr Arg Glu Lys Tyr
Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser220 225
230 235ccc aga act aac atg act gcc cac atc acg
gtc ccc tac ctt ggt gtg 1973Pro Arg Thr Asn Met Thr Ala His Ile Thr
Val Pro Tyr Leu Gly Val 240 245
250aac agg tat gat cag tac aag aag cat aag ccc tgg aca ttg gtt gtc
2021Asn Arg Tyr Asp Gln Tyr Lys Lys His Lys Pro Trp Thr Leu Val Val
255 260 265atg gtc gtg tcg cca ctt
acg gtc aac aac act agt gcg gca caa atc 2069Met Val Val Ser Pro Leu
Thr Val Asn Asn Thr Ser Ala Ala Gln Ile 270 275
280aag gtc tac gcc aac ata gct ccg acc tat gtt cac gtg gcc
ggt gaa 2117Lys Val Tyr Ala Asn Ile Ala Pro Thr Tyr Val His Val Ala
Gly Glu 285 290 295ctc ccc tcg aaa gag
ggg att ttc ccg gtt gca tgt gcg gac ggt tac 2165Leu Pro Ser Lys Glu
Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr300 305
310 315gga gga ttg gtg acg aca gac ccg aag aca
gct gac cct gct tat ggc 2213Gly Gly Leu Val Thr Thr Asp Pro Lys Thr
Ala Asp Pro Ala Tyr Gly 320 325
330aag gtg tac aac ccg cct agg act aac tac cct ggg cgc ttc acc aac
2261Lys Val Tyr Asn Pro Pro Arg Thr Asn Tyr Pro Gly Arg Phe Thr Asn
335 340 345ctg ttg gac gtg gcc gaa
gcg tgt ccc act ttc ctc tgc ttt gac gac 2309Leu Leu Asp Val Ala Glu
Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp 350 355
360ggg aaa ccg tac gtc acc acg cgg acg gat gac acc cga ctt
ttg gcc 2357Gly Lys Pro Tyr Val Thr Thr Arg Thr Asp Asp Thr Arg Leu
Leu Ala 365 370 375aag ttt gac ctt tcc
ctt gcc gca aaa cat atg tcc aac aca tac ctg 2405Lys Phe Asp Leu Ser
Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu380 385
390 395tca ggg att gct cag tac tac aca cag tac
tct ggc acc atc aat ttg 2453Ser Gly Ile Ala Gln Tyr Tyr Thr Gln Tyr
Ser Gly Thr Ile Asn Leu 400 405
410cat ttc atg ttt aca ggt tcc act gat tca aag gcc cga tac atg gtg
2501His Phe Met Phe Thr Gly Ser Thr Asp Ser Lys Ala Arg Tyr Met Val
415 420 425gcc tac atc cca cct ggg
gtg gag aca cca ccg gac aca cct gaa agg 2549Ala Tyr Ile Pro Pro Gly
Val Glu Thr Pro Pro Asp Thr Pro Glu Arg 430 435
440gct gcc cac tgc att cac gct gaa tgg gac act gga cta aac
tcc aaa 2597Ala Ala His Cys Ile His Ala Glu Trp Asp Thr Gly Leu Asn
Ser Lys 445 450 455ttc act ttc tca atc
ccg tac gta tcc gcc gcg gat tac gcg tac aca 2645Phe Thr Phe Ser Ile
Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr Thr460 465
470 475gcg tct gac acg gca gaa aca atc aac gta
cag gga tgg gtc tgc atc 2693Ala Ser Asp Thr Ala Glu Thr Ile Asn Val
Gln Gly Trp Val Cys Ile 480 485
490tac caa att aca cac ggg aag gct gaa aat gac acc ttg gtc gtg tcg
2741Tyr Gln Ile Thr His Gly Lys Ala Glu Asn Asp Thr Leu Val Val Ser
495 500 505gtt agc gcc ggc aaa gac
ttt gag ttg cgc ctc ccg att gac ccc cgc 2789Val Ser Ala Gly Lys Asp
Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg 510 515
520cag cag acc acc gct acc ggg gaa tca gca gac ccg gtc acc
acc acc 2837Gln Gln Thr Thr Ala Thr Gly Glu Ser Ala Asp Pro Val Thr
Thr Thr 525 530 535gtg gag aac tac ggc
ggt gag aca caa atc cag aga cgt cac cac acg 2885Val Glu Asn Tyr Gly
Gly Glu Thr Gln Ile Gln Arg Arg His His Thr540 545
550 555gac att ggt ttc atc atg gac aga ttt gtg
aag atc caa agc ttg agc 2933Asp Ile Gly Phe Ile Met Asp Arg Phe Val
Lys Ile Gln Ser Leu Ser 560 565
570cca aca cat gtc att gac ctc atg cag gct cac caa cac ggt ctg gtg
2981Pro Thr His Val Ile Asp Leu Met Gln Ala His Gln His Gly Leu Val
575 580 585ggt gcc ttg ctg cgt gca
gcc acg tac tac ttt tct gac ctg gaa att 3029Gly Ala Leu Leu Arg Ala
Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile 590 595
600gtt gta cgg cac gaa ggc aat ctg acc tgg gtg ccc aac ggc
gcc cct 3077Val Val Arg His Glu Gly Asn Leu Thr Trp Val Pro Asn Gly
Ala Pro 605 610 615gaa tca gcc ctg ttg
aac acc agc aac ccc act gcc tac aac aag gca 3125Glu Ser Ala Leu Leu
Asn Thr Ser Asn Pro Thr Ala Tyr Asn Lys Ala620 625
630 635cca ttc acg aga ctc gct ctc ccc tac act
gcg ccg cac cgt gtg ctg 3173Pro Phe Thr Arg Leu Ala Leu Pro Tyr Thr
Ala Pro His Arg Val Leu 640 645
650gca aca gtg tac aac ggg acg agt aag tat gct gtg ggt ggt tca ggc
3221Ala Thr Val Tyr Asn Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly
655 660 665aga aga ggc gac atg ggg
tct ctc gcg gcg cga gtc gtg aaa cag ctt 3269Arg Arg Gly Asp Met Gly
Ser Leu Ala Ala Arg Val Val Lys Gln Leu 670 675
680cct gct tca ttt aac tac ggt gca atc aag gcc gac gcc atc
cac gaa 3317Pro Ala Ser Phe Asn Tyr Gly Ala Ile Lys Ala Asp Ala Ile
His Glu 685 690 695ctt ctc gtg cgc atg
aaa cgg gcc gag ctc tac tgc ccc aga ccg ctg 3365Leu Leu Val Arg Met
Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu700 705
710 715ttg gca ata gag gtg tct tcg caa gac agg
cac aag caa aag atc att 3413Leu Ala Ile Glu Val Ser Ser Gln Asp Arg
His Lys Gln Lys Ile Ile 720 725
730gca cca gca aag cag ctt ctg aat ttt gac ctg ctc aag ttg gcc gga
3461Ala Pro Ala Lys Gln Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala Gly
735 740 745gac gtt gag tcc aac ccc
ggg cca ttc ttc ttt gct gac gtt agg tca 3509Asp Val Glu Ser Asn Pro
Gly Pro Phe Phe Phe Ala Asp Val Arg Ser 750 755
760aac ttt tca aag ttg gta gac aca atc aac cag atg cag gag
gac atg 3557Asn Phe Ser Lys Leu Val Asp Thr Ile Asn Gln Met Gln Glu
Asp Met 765 770 775tcc aca aaa cac ggg
ccc gac ttc aac cgg ttg gtg tcc gca ttt gag 3605Ser Thr Lys His Gly
Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu780 785
790 795gaa ttg gcc act gga gtt aaa gct atc agg
acc ggt ctc gac gag gcc 3653Glu Leu Ala Thr Gly Val Lys Ala Ile Arg
Thr Gly Leu Asp Glu Ala 800 805
810aaa ccc tgg tac aag ctt atc aaa ctc cta agc cgc ctg tcg tgc atg
3701Lys Pro Trp Tyr Lys Leu Ile Lys Leu Leu Ser Arg Leu Ser Cys Met
815 820 825gcc gct gtg gca gca cgg
tcc aag gac cca gtc ctt gtg gcc atc atg 3749Ala Ala Val Ala Ala Arg
Ser Lys Asp Pro Val Leu Val Ala Ile Met 830 835
840ctg gcc gac acc ggt ctc gag cgt cag aga cct ctg aaa gtg
aga gct 3797Leu Ala Asp Thr Gly Leu Glu Arg Gln Arg Pro Leu Lys Val
Arg Ala 845 850 855aag ctc cca cag cag
gaa gga cct tac gct ggc ccg ttg gag aga cag 3845Lys Leu Pro Gln Gln
Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln860 865
870 875aaa ccg ctg aaa gtg aaa gca aaa gcc ccg
gtc gtc aag gaa gga cct 3893Lys Pro Leu Lys Val Lys Ala Lys Ala Pro
Val Val Lys Glu Gly Pro 880 885
890tac gag gga ccg gtg aag aag cct gtc gct ttg aaa gtg aaa gct aag
3941Tyr Glu Gly Pro Val Lys Lys Pro Val Ala Leu Lys Val Lys Ala Lys
895 900 905aac ttg ata gtc act gag
agt ggt gcc cca ccg acc gac ttg caa aag 3989Asn Leu Ile Val Thr Glu
Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys 910 915
920atg gtc atg ggc aac aca aag cct gtt gag ctc atc ctt gac
ggg aag 4037Met Val Met Gly Asn Thr Lys Pro Val Glu Leu Ile Leu Asp
Gly Lys 925 930 935aca gta gcc atc tgt
tgt gct act gga gtg ttt ggc act gct tac ctc 4085Thr Val Ala Ile Cys
Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu940 945
950 955gtg cct cgt cat ctt ttc gca gag aag tat
gac aag atc atg ctg gat 4133Val Pro Arg His Leu Phe Ala Glu Lys Tyr
Asp Lys Ile Met Leu Asp 960 965
970ggc aga gcc atg aca gac agt gac tac aga gtg ttt gag ttt gag att
4181Gly Arg Ala Met Thr Asp Ser Asp Tyr Arg Val Phe Glu Phe Glu Ile
975 980 985aaa gta aaa gga cag gac
atg ctc tca gac gct gcg ctc atg gtg ctc 4229Lys Val Lys Gly Gln Asp
Met Leu Ser Asp Ala Ala Leu Met Val Leu 990 995
1000cac cgt ggg aac cgc gtg aga gat atc acg aaa cac ttt cgt
gat aca 4277His Arg Gly Asn Arg Val Arg Asp Ile Thr Lys His Phe Arg
Asp Thr 1005 1010 1015gca aga atg aag
aaa ggc acc ccc gtc gtc ggt gtg gtc aac aac gcc 4325Ala Arg Met Lys
Lys Gly Thr Pro Val Val Gly Val Val Asn Asn Ala1020 1025
1030 1035gac gtt ggg aga ctg att ttc tct ggt
gag gcc ctc acc tac aag gat 4373Asp Val Gly Arg Leu Ile Phe Ser Gly
Glu Ala Leu Thr Tyr Lys Asp 1040 1045
1050att gta gtg tgc atg gac gga gac acc atg cct ggc ctc ttt gcc
tac 4421Ile Val Val Cys Met Asp Gly Asp Thr Met Pro Gly Leu Phe Ala
Tyr 1055 1060 1065aaa gcc gcc
acc aag gca ggc tac tgt gga gga gcc gtt ctc gcc aag 4469Lys Ala Ala
Thr Lys Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys 1070
1075 1080gac ggg gcc gac act ttc atc gtc ggc act cac
tcc gca gga ggc aat 4517Asp Gly Ala Asp Thr Phe Ile Val Gly Thr His
Ser Ala Gly Gly Asn 1085 1090 1095gga
gtt gga tac tgc tca tgc gtt tcc agg tcc atg ctt ctc aga atg 4565Gly
Val Gly Tyr Cys Ser Cys Val Ser Arg Ser Met Leu Leu Arg Met1100
1105 1110 1115aag gca cac gtt gac cct
gaa cca caa cac gag tagtaatttt tctgcagccc 4618Lys Ala His Val Asp Pro
Glu Pro Gln His Glu 1120 1125gggtttttat
agctaattag tcattttttc gtaagtaagt atttttattt aatacttttt 4678attgtactta
tgttaaatat aactgatgat aacaaaatcc attatgtatt atttataact 4738gtaatttctt
tagcgtagtt agatgtccaa tctctctcaa atacatcggc tatcttttta 4798gtgagatttt
gatctatgca gttgaaactt atgaacgcgt gatgattaaa atgtgaaccg 4858tccaaatttg
cagtcattat atgagcgtat ctattatcta ctatcatcat ctttgagtta 4918ttaatatcat
ctactttaga attgatagga aatatgaata cctttgtagt aatatctata 4978ctatctacac
ctaactcatt aagacttttg atag
5012591126PRTFoot-and-mouth disease virus 59Met Gly Ala Gly Gln Ser Ser
Pro Ala Thr Gly Ser Gln Asn Gln Ser1 5 10
15Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr Met Gln
Gln Tyr Gln 20 25 30Asn Ser
Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly Gly Ser 35
40 45Asn Glu Gly Ser Thr Asp Thr Thr Ser Thr
His Thr Thr Asn Thr Gln 50 55 60Asn
Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu65
70 75 80Phe Gly Ala Leu Leu Ala
Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu 85
90 95Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His Thr
Thr Ser Thr Thr 100 105 110Gln
Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu Asp His 115
120 125Val Ala Gly Pro Asn Thr Ser Gly Leu
Glu Thr Arg Val Val Gln Ala 130 135
140Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys Ala145
150 155 160Phe Gly His Leu
Glu Lys Leu Glu Leu Pro Ser Asp His His Gly Val 165
170 175Phe Gly His Leu Val Asp Ser Tyr Ala Tyr
Met Arg Asn Gly Trp Asp 180 185
190Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys Leu Leu
195 200 205Val Ala Met Val Pro Glu Trp
Lys Glu Phe Asp Thr Arg Glu Lys Tyr 210 215
220Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro Arg Thr Asn
Met225 230 235 240Thr Ala
His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln
245 250 255Tyr Lys Lys His Lys Pro Trp
Thr Leu Val Val Met Val Val Ser Pro 260 265
270Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys Val Tyr
Ala Asn 275 280 285Ile Ala Pro Thr
Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu 290
295 300Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr Gly
Gly Leu Val Thr305 310 315
320Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn Pro
325 330 335Pro Arg Thr Asn Tyr
Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala 340
345 350Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly
Lys Pro Tyr Val 355 360 365Thr Thr
Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser 370
375 380Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu
Ser Gly Ile Ala Gln385 390 395
400Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe Thr
405 410 415Gly Ser Thr Asp
Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro 420
425 430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg
Ala Ala His Cys Ile 435 440 445His
Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile 450
455 460Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr
Thr Ala Ser Asp Thr Ala465 470 475
480Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr Gln Ile Thr
His 485 490 495Gly Lys Ala
Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys 500
505 510Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro
Arg Gln Gln Thr Thr Ala 515 520
525Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu Asn Tyr Gly 530
535 540Gly Glu Thr Gln Ile Gln Arg Arg
His His Thr Asp Ile Gly Phe Ile545 550
555 560Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser Pro
Thr His Val Ile 565 570
575Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
580 585 590Ala Ala Thr Tyr Tyr Phe
Ser Asp Leu Glu Ile Val Val Arg His Glu 595 600
605Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ser Ala
Leu Leu 610 615 620Asn Thr Ser Asn Pro
Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu625 630
635 640Ala Leu Pro Tyr Thr Ala Pro His Arg Val
Leu Ala Thr Val Tyr Asn 645 650
655Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly Asp Met
660 665 670Gly Ser Leu Ala Ala
Arg Val Val Lys Gln Leu Pro Ala Ser Phe Asn 675
680 685Tyr Gly Ala Ile Lys Ala Asp Ala Ile His Glu Leu
Leu Val Arg Met 690 695 700Lys Arg Ala
Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val705
710 715 720Ser Ser Gln Asp Arg His Lys
Gln Lys Ile Ile Ala Pro Ala Lys Gln 725
730 735Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp
Val Glu Ser Asn 740 745 750Pro
Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser Lys Leu 755
760 765Val Asp Thr Ile Asn Gln Met Gln Glu
Asp Met Ser Thr Lys His Gly 770 775
780Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr Gly785
790 795 800Val Lys Ala Ile
Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr Lys 805
810 815Leu Ile Lys Leu Leu Ser Arg Leu Ser Cys
Met Ala Ala Val Ala Ala 820 825
830Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp Thr Gly
835 840 845Leu Glu Arg Gln Arg Pro Leu
Lys Val Arg Ala Lys Leu Pro Gln Gln 850 855
860Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys Pro Leu Lys
Val865 870 875 880Lys Ala
Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro Val
885 890 895Lys Lys Pro Val Ala Leu Lys
Val Lys Ala Lys Asn Leu Ile Val Thr 900 905
910Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met Val Met
Gly Asn 915 920 925Thr Lys Pro Val
Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys 930
935 940Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu Val
Pro Arg His Leu945 950 955
960Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met Thr
965 970 975Asp Ser Asp Tyr Arg
Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln 980
985 990Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu His
Arg Gly Asn Arg 995 1000 1005Val Arg
Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys 1010
1015 1020Gly Thr Pro Val Val Gly Val Val Asn Asn Ala
Asp Val Gly Arg Leu1025 1030 1035
1040Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys Met
1045 1050 1055Asp Gly Asp Thr
Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys 1060
1065 1070Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys
Asp Gly Ala Asp Thr 1075 1080
1085Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val Gly Tyr Cys
1090 1095 1100Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met Lys Ala His Val Asp1105 1110
1115 1120Pro Glu Pro Gln His Glu
112560111DNAArtificial SequenceDescription of Artificial Sequence
Synthetic oligonucleotide 60gaattcactg taaaaataga aactataatc
atataatagt gtaggttggt agtagggtac 60tcgtgattaa ttttattgtt aaacttgatg
ggagctgggc aatccagccc a 11161105DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
61tgggctggat tgcccagctc ccatcaagtt taacaataaa attaatcacg agtaccctac
60taccaaccta cactattata tgattatagt ttctattttt acagt
1056222DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 62cttgccgcaa aacatatgtc ca
226322DNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 63tggacatatg ttttgcggca ag
226427DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
64actgtaaaaa tagaaactat aatcata
276527DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 65taatagtgta ggttggtagt agggtac
276627DNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 66tcgtgattaa ttttattgtt aaacttg
276740DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
67acctacacta ttatatgatt atagtttcta tttttacagt
406841DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 68caagtttaac aataaaatta atcacgagta ccctactacc a
416923DNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 69actgtaaaaa tagaaactat aat
237024DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
70caagtttaac aataaaatta atca
2471111DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 71gaattcactg taaaaataga aactataatc atataatagt
gtaggttggt agtagggtac 60tcgtgattaa ttttattgtt aaacttgatg ggagctgggc
aatccagccc a 11172105DNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 72tgggctggat
tgcccagctc ccatcaagtt taacaataaa attaatcacg agtaccctac 60taccaaccta
cactattata tgattatagt ttctattttt acagt
1057326DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 73gaattcactg taaaaataga aactat
267428DNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 74cagctcccat caagtttaac aataaaat
287528DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
75gttaaacttg atgggagctg ggcaatcc
287623DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 76tgcctcctgg tggccatggt acc
237723DNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 77ggtaccatgg ccaccaggag gca
23788PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 78Met
Gly Ala Gly Gln Ser Ser Pro1 5797PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 79Leu
Ala Ala Lys His Met Ser1 5808PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 80Cys
Leu Leu Val Ala Met Val Pro1 5815059DNAFoot-and-mouth
disease virusCDS(1268)..(4645) 81ttcataaata caagtttgat taaacttaag
ttgttctaaa gttctttcct ccgaaggtat 60agaacaaagt atttcttcta catccttact
atttattgca gcttttaaca gcctatcacg 120tatcctattt ttagtattgg tagaacgttt
tagttctaaa gttaaaatat tagacataat 180tggcatattg cttattcctt gcatagttga
gtctgtagat cgtttcagta tatcactgat 240taatgtacta ctgttatgat gaaatataga
atcgatattg gcatttaact gttttgttat 300actaagtcta gattttaaat cttctagtaa
tatgctattt aatataaaag cttccacgtt 360tttgtataca tttctttcca tattagtagc
tactactaaa tgattatctt ctttcatatc 420ttgtagataa gatagactat ctttatcttt
attagtagaa aatacttctg gccatacatc 480gttaaatttt tttgttgttg ttagatataa
tattaaatat ctagaggatc ctattatttg 540tggtaaaatg tttatagagt aaaatgatct
ggctattaaa cataggccag ttaccataga 600atgctgcttc ccgttacagt gttttaccat
aaccatagat ctgcctgtat tgttgataca 660tataacagct gtaaatccta aaaaattcct
atcataatta ttaatattag gtaattcatt 720tccatgtgaa agatagacta attttatatc
ctttacctcc aaataattat ttacatctct 780taaacaatct attttaatat cattaactgg
tattttataa tatccagaaa ggtttgaagg 840ggttgatgga ataagtctat taacatcgtt
aagtaaatta ttaatatcat gaatctttat 900tatattatac ccataagtta aatttatatt
tactttctca tcatctgact tagttagttt 960gtaataaggt gtgtctgaaa aaattaaaag
gtaattcgtt gaatgaagct gtatttgctg 1020tatcattttt atctaatttt ggagatttag
cagtacttac ttcattagaa gaagaatctg 1080ccagttcctg tctattactg atatttcgtt
tcattattat atgatttata ttttactttt 1140tcaattatat atactcattt gactagttaa
tcaataaaaa gaattcactg taaaaataga 1200aactataatc atataatagt gtaggttggt
agtagggtac tcgtgattaa ttttattgtt 1260aaacttg atg gga gct ggg caa tcc
agc cca gca acc ggc tcg cag aac 1309 Met Gly Ala Gly Gln Ser
Ser Pro Ala Thr Gly Ser Gln Asn 1 5
10cag tct ggc aac act ggc agc ata atc aac aac tac tac atg caa cag
1357Gln Ser Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr Tyr Met Gln Gln15
20 25 30tac cag aac tcc
atg gac aca cag ttg gga gac aat gcc atc agt gga 1405Tyr Gln Asn Ser
Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly 35
40 45ggc tcc aac gag ggc tcc acg gac aca act
tca aca cac aca acc aac 1453Gly Ser Asn Glu Gly Ser Thr Asp Thr Thr
Ser Thr His Thr Thr Asn 50 55
60act caa aac aat gac tgg ttc tcg aag ctc gcc agt tca gct ttt acc
1501Thr Gln Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr
65 70 75ggt ctg ttc ggt gca ctg ctc
gcc gac aag aag aca gag gaa acg aca 1549Gly Leu Phe Gly Ala Leu Leu
Ala Asp Lys Lys Thr Glu Glu Thr Thr 80 85
90ctt ctt gag gac cgc atc ctc acc acc cgc aac ggg cac acc acc tcg
1597Leu Leu Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His Thr Thr Ser95
100 105 110acg acc caa tcg
agt gtg ggt gtc aca cac ggg tac tcc aca gag gag 1645Thr Thr Gln Ser
Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu 115
120 125gac cac gtt gct ggg ccc aac aca tcg ggc
ctg gag acg cga gtg gtg 1693Asp His Val Ala Gly Pro Asn Thr Ser Gly
Leu Glu Thr Arg Val Val 130 135
140cag gca gag aga ttc tac aaa aag tac ttg ttt gac tgg aca acg gac
1741Gln Ala Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp
145 150 155aag gca ttt gga cac ctg gaa
aag ctg gag ctc ccg tcc gac cac cac 1789Lys Ala Phe Gly His Leu Glu
Lys Leu Glu Leu Pro Ser Asp His His 160 165
170ggt gtc ttt gga cac ttg gtg gac tcg tac gcc tat atg aga aat ggc
1837Gly Val Phe Gly His Leu Val Asp Ser Tyr Ala Tyr Met Arg Asn Gly175
180 185 190tgg gat gtt gag
gtg tcc gct gtt ggc aac cag ttc aac ggc ggg tgc 1885Trp Asp Val Glu
Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys 195
200 205ctc ctg gtg gcc atg gta cct gaa tgg aag
gaa ttt gac aca cgg gag 1933Leu Leu Val Ala Met Val Pro Glu Trp Lys
Glu Phe Asp Thr Arg Glu 210 215
220aaa tac caa ctc acc ctt ttc ccg cac cag ttt att agc ccc aga act
1981Lys Tyr Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro Arg Thr
225 230 235aac atg act gcc cac atc acg
gtc ccc tac ctt ggt gtg aac agg tat 2029Asn Met Thr Ala His Ile Thr
Val Pro Tyr Leu Gly Val Asn Arg Tyr 240 245
250gat cag tac aag aag cat aag ccc tgg aca ttg gtt gtc atg gtc gtg
2077Asp Gln Tyr Lys Lys His Lys Pro Trp Thr Leu Val Val Met Val Val255
260 265 270tcg cca ctt acg
gtc aac aac act agt gcg gca caa atc aag gtc tac 2125Ser Pro Leu Thr
Val Asn Asn Thr Ser Ala Ala Gln Ile Lys Val Tyr 275
280 285gcc aac ata gct ccg acc tat gtt cac gtg
gcc ggt gaa ctc ccc tcg 2173Ala Asn Ile Ala Pro Thr Tyr Val His Val
Ala Gly Glu Leu Pro Ser 290 295
300aaa gag ggg att ttc ccg gtt gca tgt gcg gac ggt tac gga gga ttg
2221Lys Glu Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr Gly Gly Leu
305 310 315gtg acg aca gac ccg aag aca
gct gac cct gct tat ggc aag gtg tac 2269Val Thr Thr Asp Pro Lys Thr
Ala Asp Pro Ala Tyr Gly Lys Val Tyr 320 325
330aac ccg cct agg act aac tac cct ggg cgc ttc acc aac ctg ttg gac
2317Asn Pro Pro Arg Thr Asn Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp335
340 345 350gtg gcc gaa gcg
tgt ccc act ttc ctc tgc ttt gac gac ggg aaa ccg 2365Val Ala Glu Ala
Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly Lys Pro 355
360 365tac gtc acc acg cgg acg gat gac acc cga
ctt ttg gcc aag ttt gac 2413Tyr Val Thr Thr Arg Thr Asp Asp Thr Arg
Leu Leu Ala Lys Phe Asp 370 375
380ctt tcc ctt gcc gca aaa cat atg tcc aac aca tac ctg tca ggg att
2461Leu Ser Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu Ser Gly Ile
385 390 395gct cag tac tac aca cag tac
tct ggc acc atc aat ttg cat ttc atg 2509Ala Gln Tyr Tyr Thr Gln Tyr
Ser Gly Thr Ile Asn Leu His Phe Met 400 405
410ttt aca ggt tcc act gat tca aag gcc cga tac atg gtg gcc tac atc
2557Phe Thr Gly Ser Thr Asp Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile415
420 425 430cca cct ggg gtg
gag aca cca ccg gac aca cct gaa agg gct gcc cac 2605Pro Pro Gly Val
Glu Thr Pro Pro Asp Thr Pro Glu Arg Ala Ala His 435
440 445tgc att cac gct gaa tgg gac act gga cta
aac tcc aaa ttc act ttc 2653Cys Ile His Ala Glu Trp Asp Thr Gly Leu
Asn Ser Lys Phe Thr Phe 450 455
460tca atc ccg tac gta tcc gcc gcg gat tac gcg tac aca gcg tct gac
2701Ser Ile Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr Thr Ala Ser Asp
465 470 475acg gca gaa aca atc aac gta
cag gga tgg gtc tgc atc tac caa att 2749Thr Ala Glu Thr Ile Asn Val
Gln Gly Trp Val Cys Ile Tyr Gln Ile 480 485
490aca cac ggg aag gct gaa aat gac acc ttg gtc gtg tcg gtt agc gcc
2797Thr His Gly Lys Ala Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala495
500 505 510ggc aaa gac ttt
gag ttg cgc ctc ccg att gac ccc cgc cag cag acc 2845Gly Lys Asp Phe
Glu Leu Arg Leu Pro Ile Asp Pro Arg Gln Gln Thr 515
520 525acc gct acc ggg gaa tca gca gac ccg gtc
acc acc acc gtg gag aac 2893Thr Ala Thr Gly Glu Ser Ala Asp Pro Val
Thr Thr Thr Val Glu Asn 530 535
540tac ggc ggt gag aca caa atc cag aga cgt cac cac acg gac att ggt
2941Tyr Gly Gly Glu Thr Gln Ile Gln Arg Arg His His Thr Asp Ile Gly
545 550 555ttc atc atg gac aga ttt gtg
aag atc caa agc ttg agc cca aca cat 2989Phe Ile Met Asp Arg Phe Val
Lys Ile Gln Ser Leu Ser Pro Thr His 560 565
570gtc att gac ctc atg cag gct cac caa cac ggt ctg gtg ggt gcc ttg
3037Val Ile Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu575
580 585 590ctg cgt gca gcc
acg tac tac ttt tct gac ctg gaa att gtt gta cgg 3085Leu Arg Ala Ala
Thr Tyr Tyr Phe Ser Asp Leu Glu Ile Val Val Arg 595
600 605cac gaa ggc aat ctg acc tgg gtg ccc aac
ggc gcc cct gaa tca gcc 3133His Glu Gly Asn Leu Thr Trp Val Pro Asn
Gly Ala Pro Glu Ser Ala 610 615
620ctg ttg aac acc agc aac ccc act gcc tac aac aag gca cca ttc acg
3181Leu Leu Asn Thr Ser Asn Pro Thr Ala Tyr Asn Lys Ala Pro Phe Thr
625 630 635aga ctc gct ctc ccc tac act
gcg ccg cac cgt gtg ctg gca aca gtg 3229Arg Leu Ala Leu Pro Tyr Thr
Ala Pro His Arg Val Leu Ala Thr Val 640 645
650tac aac ggg acg agt aag tat gct gtg ggt ggt tca ggc aga aga ggc
3277Tyr Asn Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly655
660 665 670gac atg ggg tct
ctc gcg gcg cga gtc gtg aaa cag ctt cct gct tca 3325Asp Met Gly Ser
Leu Ala Ala Arg Val Val Lys Gln Leu Pro Ala Ser 675
680 685ttt aac tac ggt gca atc aag gcc gac gcc
atc cac gaa ctt ctc gtg 3373Phe Asn Tyr Gly Ala Ile Lys Ala Asp Ala
Ile His Glu Leu Leu Val 690 695
700cgc atg aaa cgg gcc gag ctc tac tgc ccc aga ccg ctg ttg gca ata
3421Arg Met Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile
705 710 715gag gtg tct tcg caa gac agg
cac aag caa aag atc att gca cca gca 3469Glu Val Ser Ser Gln Asp Arg
His Lys Gln Lys Ile Ile Ala Pro Ala 720 725
730aag cag ctt ctg aat ttt gac ctg ctc aag ttg gcc gga gac gtt gag
3517Lys Gln Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp Val Glu735
740 745 750tcc aac ccc ggg
cca ttc ttc ttt gct gac gtt agg tca aac ttt tca 3565Ser Asn Pro Gly
Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser 755
760 765aag ttg gta gac aca atc aac cag atg cag
gag gac atg tcc aca aaa 3613Lys Leu Val Asp Thr Ile Asn Gln Met Gln
Glu Asp Met Ser Thr Lys 770 775
780cac ggg ccc gac ttc aac cgg ttg gtg tcc gca ttt gag gaa ttg gcc
3661His Gly Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala
785 790 795act gga gtt aaa gct atc agg
acc ggt ctc gac gag gcc aaa ccc tgg 3709Thr Gly Val Lys Ala Ile Arg
Thr Gly Leu Asp Glu Ala Lys Pro Trp 800 805
810tac aag ctt atc aaa ctc cta agc cgc ctg tcg tgc atg gcc gct gtg
3757Tyr Lys Leu Ile Lys Leu Leu Ser Arg Leu Ser Cys Met Ala Ala Val815
820 825 830gca gca cgg tcc
aag gac cca gtc ctt gtg gcc atc atg ctg gcc gac 3805Ala Ala Arg Ser
Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp 835
840 845acc ggt ctc gag cgt cag aga cct ctg aaa
gtg aga gct aag ctc cca 3853Thr Gly Leu Glu Arg Gln Arg Pro Leu Lys
Val Arg Ala Lys Leu Pro 850 855
860cag cag gaa gga cct tac gct ggc ccg ttg gag aga cag aaa ccg ctg
3901Gln Gln Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys Pro Leu
865 870 875aaa gtg aaa gca aaa gcc ccg
gtc gtc aag gaa gga cct tac gag gga 3949Lys Val Lys Ala Lys Ala Pro
Val Val Lys Glu Gly Pro Tyr Glu Gly 880 885
890ccg gtg aag aag cct gtc gct ttg aaa gtg aaa gct aag aac ttg ata
3997Pro Val Lys Lys Pro Val Ala Leu Lys Val Lys Ala Lys Asn Leu Ile895
900 905 910gtc act gag agt
ggt gcc cca ccg acc gac ttg caa aag atg gtc atg 4045Val Thr Glu Ser
Gly Ala Pro Pro Thr Asp Leu Gln Lys Met Val Met 915
920 925ggc aac aca aag cct gtt gag ctc atc ctt
gac ggg aag aca gta gcc 4093Gly Asn Thr Lys Pro Val Glu Leu Ile Leu
Asp Gly Lys Thr Val Ala 930 935
940atc tgt tgt gct act gga gtg ttt ggc act gct tac ctc gtg cct cgt
4141Ile Cys Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu Val Pro Arg
945 950 955cat ctt ttc gca gag aag tat
gac aag atc atg ctg gat ggc aga gcc 4189His Leu Phe Ala Glu Lys Tyr
Asp Lys Ile Met Leu Asp Gly Arg Ala 960 965
970atg aca gac agt gac tac aga gtg ttt gag ttt gag att aaa gta aaa
4237Met Thr Asp Ser Asp Tyr Arg Val Phe Glu Phe Glu Ile Lys Val Lys975
980 985 990gga cag gac atg
ctc tca gac gct gcg ctc atg gtg ctc cac cgt ggg 4285Gly Gln Asp Met
Leu Ser Asp Ala Ala Leu Met Val Leu His Arg Gly 995
1000 1005aac cgc gtg aga gat atc acg aaa cac ttt
cgt gat aca gca aga atg 4333Asn Arg Val Arg Asp Ile Thr Lys His Phe
Arg Asp Thr Ala Arg Met 1010 1015
1020aag aaa ggc acc ccc gtc gtc ggt gtg gtc aac aac gcc gac gtt ggg
4381Lys Lys Gly Thr Pro Val Val Gly Val Val Asn Asn Ala Asp Val Gly
1025 1030 1035aga ctg att ttc tct ggt gag
gcc ctc acc tac aag gat att gta gtg 4429Arg Leu Ile Phe Ser Gly Glu
Ala Leu Thr Tyr Lys Asp Ile Val Val 1040 1045
1050tgc atg gac gga gac acc atg cct ggc ctc ttt gcc tac aaa gcc gcc
4477Cys Met Asp Gly Asp Thr Met Pro Gly Leu Phe Ala Tyr Lys Ala
Ala1055 1060 1065 1070acc aag
gca ggc tac tgt gga gga gcc gtt ctc gcc aag gac ggg gcc 4525Thr Lys
Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys Asp Gly Ala
1075 1080 1085gac act ttc atc gtc ggc act
cac tcc gca gga ggc aat gga gtt gga 4573Asp Thr Phe Ile Val Gly Thr
His Ser Ala Gly Gly Asn Gly Val Gly 1090 1095
1100tac tgc tca tgc gtt tcc agg tcc atg ctt ctc aga atg aag
gca cac 4621Tyr Cys Ser Cys Val Ser Arg Ser Met Leu Leu Arg Met Lys
Ala His 1105 1110 1115gtt gac cct
gaa cca caa cac gag tagtaatttt tctgcagccc gggtttttat 4675Val Asp Pro
Glu Pro Gln His Glu 1120 1125agctaattag tcattttttc
gtaagtaagt atttttattt aatacttttt attgtactta 4735tgttaaatat aactgatgat
aacaaaatcc attatgtatt atttataact gtaatttctt 4795tagcgtagtt agatgtccaa
tctctctcaa atacatcggc tatcttttta gtgagatttt 4855gatctatgca gttgaaactt
atgaacgcgt gatgattaaa atgtgaaccg tccaaatttg 4915cagtcattat atgagcgtat
ctattatcta ctatcatcat ctttgagtta ttaatatcat 4975ctactttaga attgatagga
aatatgaata cctttgtagt aatatctata ctatctacac 5035ctaactcatt aagacttttg
atag 5059821126PRTFoot-and-mouth
disease virus 82Met Gly Ala Gly Gln Ser Ser Pro Ala Thr Gly Ser Gln Asn
Gln Ser1 5 10 15Gly Asn
Thr Gly Ser Ile Ile Asn Asn Tyr Tyr Met Gln Gln Tyr Gln 20
25 30Asn Ser Met Asp Thr Gln Leu Gly Asp
Asn Ala Ile Ser Gly Gly Ser 35 40
45Asn Glu Gly Ser Thr Asp Thr Thr Ser Thr His Thr Thr Asn Thr Gln 50
55 60Asn Asn Asp Trp Phe Ser Lys Leu Ala
Ser Ser Ala Phe Thr Gly Leu65 70 75
80Phe Gly Ala Leu Leu Ala Asp Lys Lys Thr Glu Glu Thr Thr
Leu Leu 85 90 95Glu Asp
Arg Ile Leu Thr Thr Arg Asn Gly His Thr Thr Ser Thr Thr 100
105 110Gln Ser Ser Val Gly Val Thr His Gly
Tyr Ser Thr Glu Glu Asp His 115 120
125Val Ala Gly Pro Asn Thr Ser Gly Leu Glu Thr Arg Val Val Gln Ala
130 135 140Glu Arg Phe Tyr Lys Lys Tyr
Leu Phe Asp Trp Thr Thr Asp Lys Ala145 150
155 160Phe Gly His Leu Glu Lys Leu Glu Leu Pro Ser Asp
His His Gly Val 165 170
175Phe Gly His Leu Val Asp Ser Tyr Ala Tyr Met Arg Asn Gly Trp Asp
180 185 190Val Glu Val Ser Ala Val
Gly Asn Gln Phe Asn Gly Gly Cys Leu Leu 195 200
205Val Ala Met Val Pro Glu Trp Lys Glu Phe Asp Thr Arg Glu
Lys Tyr 210 215 220Gln Leu Thr Leu Phe
Pro His Gln Phe Ile Ser Pro Arg Thr Asn Met225 230
235 240Thr Ala His Ile Thr Val Pro Tyr Leu Gly
Val Asn Arg Tyr Asp Gln 245 250
255Tyr Lys Lys His Lys Pro Trp Thr Leu Val Val Met Val Val Ser Pro
260 265 270Leu Thr Val Asn Asn
Thr Ser Ala Ala Gln Ile Lys Val Tyr Ala Asn 275
280 285Ile Ala Pro Thr Tyr Val His Val Ala Gly Glu Leu
Pro Ser Lys Glu 290 295 300Gly Ile Phe
Pro Val Ala Cys Ala Asp Gly Tyr Gly Gly Leu Val Thr305
310 315 320Thr Asp Pro Lys Thr Ala Asp
Pro Ala Tyr Gly Lys Val Tyr Asn Pro 325
330 335Pro Arg Thr Asn Tyr Pro Gly Arg Phe Thr Asn Leu
Leu Asp Val Ala 340 345 350Glu
Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly Lys Pro Tyr Val 355
360 365Thr Thr Arg Thr Asp Asp Thr Arg Leu
Leu Ala Lys Phe Asp Leu Ser 370 375
380Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu Ser Gly Ile Ala Gln385
390 395 400Tyr Tyr Thr Gln
Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe Thr 405
410 415Gly Ser Thr Asp Ser Lys Ala Arg Tyr Met
Val Ala Tyr Ile Pro Pro 420 425
430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg Ala Ala His Cys Ile
435 440 445His Ala Glu Trp Asp Thr Gly
Leu Asn Ser Lys Phe Thr Phe Ser Ile 450 455
460Pro Tyr Val Ser Ala Ala Asp Tyr Ala Tyr Thr Ala Ser Asp Thr
Ala465 470 475 480Glu Thr
Ile Asn Val Gln Gly Trp Val Cys Ile Tyr Gln Ile Thr His
485 490 495Gly Lys Ala Glu Asn Asp Thr
Leu Val Val Ser Val Ser Ala Gly Lys 500 505
510Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg Gln Gln Thr
Thr Ala 515 520 525Thr Gly Glu Ser
Ala Asp Pro Val Thr Thr Thr Val Glu Asn Tyr Gly 530
535 540Gly Glu Thr Gln Ile Gln Arg Arg His His Thr Asp
Ile Gly Phe Ile545 550 555
560Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser Pro Thr His Val Ile
565 570 575Asp Leu Met Gln Ala
His Gln His Gly Leu Val Gly Ala Leu Leu Arg 580
585 590Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile Val
Val Arg His Glu 595 600 605Gly Asn
Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ser Ala Leu Leu 610
615 620Asn Thr Ser Asn Pro Thr Ala Tyr Asn Lys Ala
Pro Phe Thr Arg Leu625 630 635
640Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala Thr Val Tyr Asn
645 650 655Gly Thr Ser Lys
Tyr Ala Val Gly Gly Ser Gly Arg Arg Gly Asp Met 660
665 670Gly Ser Leu Ala Ala Arg Val Val Lys Gln Leu
Pro Ala Ser Phe Asn 675 680 685Tyr
Gly Ala Ile Lys Ala Asp Ala Ile His Glu Leu Leu Val Arg Met 690
695 700Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro
Leu Leu Ala Ile Glu Val705 710 715
720Ser Ser Gln Asp Arg His Lys Gln Lys Ile Ile Ala Pro Ala Lys
Gln 725 730 735Leu Leu Asn
Phe Asp Leu Leu Lys Leu Ala Gly Asp Val Glu Ser Asn 740
745 750Pro Gly Pro Phe Phe Phe Ala Asp Val Arg
Ser Asn Phe Ser Lys Leu 755 760
765Val Asp Thr Ile Asn Gln Met Gln Glu Asp Met Ser Thr Lys His Gly 770
775 780Pro Asp Phe Asn Arg Leu Val Ser
Ala Phe Glu Glu Leu Ala Thr Gly785 790
795 800Val Lys Ala Ile Arg Thr Gly Leu Asp Glu Ala Lys
Pro Trp Tyr Lys 805 810
815Leu Ile Lys Leu Leu Ser Arg Leu Ser Cys Met Ala Ala Val Ala Ala
820 825 830Arg Ser Lys Asp Pro Val
Leu Val Ala Ile Met Leu Ala Asp Thr Gly 835 840
845Leu Glu Arg Gln Arg Pro Leu Lys Val Arg Ala Lys Leu Pro
Gln Gln 850 855 860Glu Gly Pro Tyr Ala
Gly Pro Leu Glu Arg Gln Lys Pro Leu Lys Val865 870
875 880Lys Ala Lys Ala Pro Val Val Lys Glu Gly
Pro Tyr Glu Gly Pro Val 885 890
895Lys Lys Pro Val Ala Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr
900 905 910Glu Ser Gly Ala Pro
Pro Thr Asp Leu Gln Lys Met Val Met Gly Asn 915
920 925Thr Lys Pro Val Glu Leu Ile Leu Asp Gly Lys Thr
Val Ala Ile Cys 930 935 940Cys Ala Thr
Gly Val Phe Gly Thr Ala Tyr Leu Val Pro Arg His Leu945
950 955 960Phe Ala Glu Lys Tyr Asp Lys
Ile Met Leu Asp Gly Arg Ala Met Thr 965
970 975Asp Ser Asp Tyr Arg Val Phe Glu Phe Glu Ile Lys
Val Lys Gly Gln 980 985 990Asp
Met Leu Ser Asp Ala Ala Leu Met Val Leu His Arg Gly Asn Arg 995
1000 1005Val Arg Asp Ile Thr Lys His Phe Arg
Asp Thr Ala Arg Met Lys Lys 1010 1015
1020Gly Thr Pro Val Val Gly Val Val Asn Asn Ala Asp Val Gly Arg Leu1025
1030 1035 1040Ile Phe Ser Gly
Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys Met 1045
1050 1055Asp Gly Asp Thr Met Pro Gly Leu Phe Ala
Tyr Lys Ala Ala Thr Lys 1060 1065
1070Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys Asp Gly Ala Asp Thr
1075 1080 1085Phe Ile Val Gly Thr His Ser
Ala Gly Gly Asn Gly Val Gly Tyr Cys 1090 1095
1100Ser Cys Val Ser Arg Ser Met Leu Leu Arg Met Lys Ala His Val
Asp1105 1110 1115 1120Pro Glu
Pro Gln His Glu 11258329DNAArtificial SequenceDescription
of Artificial Sequence Synthetic primer 83aagcttttct ttattctata
cttaaaaag 298425DNAArtificial
SequenceDescription of Artificial Sequence Synthetic primer
84ggtggccatg gtacctgaat ggaag
258525DNAArtificial SequenceDescription of Artificial Sequence Synthetic
primer 85cttccattca ggtaccatgg ccacc
25868PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 86Val Ala Met Val Pro Glu Trp Lys1
5876315DNAFoot-and-mouth disease virusCDS(1513)..(4890) 87gaccctttac
aagaataaaa gaagaaacaa ctgtgaaata gtttataaat gtaattcgta 60tgcagaaaac
gataatatat tttggtatga gaaatctaaa ggagacatag tttgtataga 120catgcgctct
tccgatgaga tattcgatgc ttttctaatg tatcatatag ctacaagata 180tgcctatcat
gatgatgata tatatctaca aatagtgtta tattattcta ataatcaaaa 240tgttatatct
tatattacga aaaataaata cgttaagtat ataagaaata aaactagaga 300cgatattcat
aaagtaaaaa tattagctct agaagacttt acaacggaag aaatatattg 360ttggattagt
aatatataac agcgtagctg cacggttttg atcattttcc aacaatataa 420accaatgaag
gaggacgact catcaaacat aaataacatt cacggaaaat attcagtatc 480agatttatca
caagatgatt atgttattga atgtatagac ggatcttttg attcgatcaa 540gtatagagat
ataaaggtta taataatgaa gaataacggt tacgttaatt gtagtaaatt 600atgtaaaatg
cggaataaat acttttctag atggttgcgt ctttctactt ctaaagcatt 660attagacatt
tacaataata agtcagtaga taatgctatt gttaaagtct atggtaaagg 720taagaaactt
attataacag gattttatct caaacaaaat atgatacgtt atgttattga 780gtggataggg
gatgatttta caaacgatat atacaaaatg attaatttct ataatgcgtt 840attcggtaac
gatgaattaa aaatagtatc ctgtgaaaac actctatgcc cgtttataga 900acttggtaga
tgctattatg gtaaaaaatg taagtatata cacggagatc aatgtgatat 960ctgtggtcta
tatatactac accctaccga tattaaccaa cgagtttctc acaagaaaac 1020ttgtttagta
gatagagatt ctttgattgt gtttaaaaga agtaccagta aaaagtgtgg 1080catatgcata
gaagaaataa acaaaaaaca tatttccgaa cagtattttg gaattctccc 1140aagttgtaaa
catatttttt gcctatcatg tataagacgt tgggcagata ctaccagaaa 1200tacagatact
gaaaatacgt gtcctgaatg tagaatagtt tttcctttca taatacccag 1260taggtattgg
atagataata aatatgataa aaaaatatta tataatagat ataagaaaat 1320gatttttaca
aaaataccta taagaacaat aaaaatataa ttacatttac ggaaaatagc 1380tggttttagt
ttaccaactt agagtaatta tcatattgaa tctatattgc taattagcta 1440ataaaaaccc
gggtcgcgaa agcttttctt tattctatac ttaaaaagtg caaataaata 1500caaaggttct
tg atg gga gct ggg caa tcc agc cca gca acc ggc tcg cag 1551
Met Gly Ala Gly Gln Ser Ser Pro Ala Thr Gly Ser Gln 1
5 10aac cag tct ggc aac act ggc agc ata atc aac
aac tac tac atg caa 1599Asn Gln Ser Gly Asn Thr Gly Ser Ile Ile Asn
Asn Tyr Tyr Met Gln 15 20 25cag tac
cag aac tcc atg gac aca cag ttg gga gac aat gcc atc agt 1647Gln Tyr
Gln Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser30
35 40 45gga ggc tcc aac gag ggc tcc
acg gac aca act tca aca cac aca acc 1695Gly Gly Ser Asn Glu Gly Ser
Thr Asp Thr Thr Ser Thr His Thr Thr 50 55
60aac act caa aac aat gac tgg ttc tcg aag ctc gcc agt
tca gct ttt 1743Asn Thr Gln Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser
Ser Ala Phe 65 70 75acc ggt
ctg ttc ggt gca ctg ctc gcc gac aag aag aca gag gaa acg 1791Thr Gly
Leu Phe Gly Ala Leu Leu Ala Asp Lys Lys Thr Glu Glu Thr 80
85 90aca ctt ctt gag gac cgc atc ctc acc acc
cgc aac ggg cac acc acc 1839Thr Leu Leu Glu Asp Arg Ile Leu Thr Thr
Arg Asn Gly His Thr Thr 95 100 105tcg
acg acc caa tcg agt gtg ggt gtc aca cac ggg tac tcc aca gag 1887Ser
Thr Thr Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu110
115 120 125gag gac cac gtt gct ggg
ccc aac aca tcg ggc ctg gag acg cga gtg 1935Glu Asp His Val Ala Gly
Pro Asn Thr Ser Gly Leu Glu Thr Arg Val 130
135 140gtg cag gca gag aga ttc tac aaa aag tac ttg ttt
gac tgg aca acg 1983Val Gln Ala Glu Arg Phe Tyr Lys Lys Tyr Leu Phe
Asp Trp Thr Thr 145 150 155gac
aag gca ttt gga cac ctg gaa aag ctg gag ctc ccg tcc gac cac 2031Asp
Lys Ala Phe Gly His Leu Glu Lys Leu Glu Leu Pro Ser Asp His 160
165 170cac ggt gtc ttt gga cac ttg gtg gac
tcg tac gcc tat atg aga aat 2079His Gly Val Phe Gly His Leu Val Asp
Ser Tyr Ala Tyr Met Arg Asn 175 180
185ggc tgg gat gtt gag gtg tcc gct gtt ggc aac cag ttc aac ggc ggg
2127Gly Trp Asp Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly190
195 200 205tgc ctc ctg gtg
gcc atg gta cct gaa tgg aag gaa ttt gac aca cgg 2175Cys Leu Leu Val
Ala Met Val Pro Glu Trp Lys Glu Phe Asp Thr Arg 210
215 220gag aaa tac caa ctc acc ctt ttc ccg cac
cag ttt att agc ccc aga 2223Glu Lys Tyr Gln Leu Thr Leu Phe Pro His
Gln Phe Ile Ser Pro Arg 225 230
235act aac atg act gcc cac atc acg gtc ccc tac ctt ggt gtg aac agg
2271Thr Asn Met Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg
240 245 250tat gat cag tac aag aag cat
aag ccc tgg aca ttg gtt gtc atg gtc 2319Tyr Asp Gln Tyr Lys Lys His
Lys Pro Trp Thr Leu Val Val Met Val 255 260
265gtg tcg cca ctt acg gtc aac aac act agt gcg gca caa atc aag gtc
2367Val Ser Pro Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys Val270
275 280 285tac gcc aac ata
gct ccg acc tat gtt cac gtg gcc ggt gaa ctc ccc 2415Tyr Ala Asn Ile
Ala Pro Thr Tyr Val His Val Ala Gly Glu Leu Pro 290
295 300tcg aaa gag ggg att ttc ccg gtt gca tgt
gcg gac ggt tac gga gga 2463Ser Lys Glu Gly Ile Phe Pro Val Ala Cys
Ala Asp Gly Tyr Gly Gly 305 310
315ttg gtg acg aca gac ccg aag aca gct gac cct gct tat ggc aag gtg
2511Leu Val Thr Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val
320 325 330tac aac ccg cct agg act aac
tac cct ggg cgc ttc acc aac ctg ttg 2559Tyr Asn Pro Pro Arg Thr Asn
Tyr Pro Gly Arg Phe Thr Asn Leu Leu 335 340
345gac gtg gcc gaa gcg tgt ccc act ttc ctc tgc ttt gac gac ggg aaa
2607Asp Val Ala Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly Lys350
355 360 365ccg tac gtc acc
acg cgg acg gat gac acc cga ctt ttg gcc aag ttt 2655Pro Tyr Val Thr
Thr Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe 370
375 380gac ctt tcc ctt gcc gca aaa cat atg tcc
aac aca tac ctg tca ggg 2703Asp Leu Ser Leu Ala Ala Lys His Met Ser
Asn Thr Tyr Leu Ser Gly 385 390
395att gct cag tac tac aca cag tac tct ggc acc atc aat ttg cat ttc
2751Ile Ala Gln Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe
400 405 410atg ttt aca ggt tcc act gat
tca aag gcc cga tac atg gtg gcc tac 2799Met Phe Thr Gly Ser Thr Asp
Ser Lys Ala Arg Tyr Met Val Ala Tyr 415 420
425atc cca cct ggg gtg gag aca cca ccg gac aca cct gaa agg gct gcc
2847Ile Pro Pro Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg Ala Ala430
435 440 445cac tgc att cac
gct gaa tgg gac act gga cta aac tcc aaa ttc act 2895His Cys Ile His
Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr 450
455 460ttc tca atc ccg tac gta tcc gcc gcg gat
tac gcg tac aca gcg tct 2943Phe Ser Ile Pro Tyr Val Ser Ala Ala Asp
Tyr Ala Tyr Thr Ala Ser 465 470
475gac acg gca gaa aca atc aac gta cag gga tgg gtc tgc atc tac caa
2991Asp Thr Ala Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr Gln
480 485 490att aca cac ggg aag gct gaa
aat gac acc ttg gtc gtg tcg gtt agc 3039Ile Thr His Gly Lys Ala Glu
Asn Asp Thr Leu Val Val Ser Val Ser 495 500
505gcc ggc aaa gac ttt gag ttg cgc ctc ccg att gac ccc cgc cag cag
3087Ala Gly Lys Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg Gln Gln510
515 520 525acc acc gct acc
ggg gaa tca gca gac ccg gtc acc acc acc gtg gag 3135Thr Thr Ala Thr
Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu 530
535 540aac tac ggc ggt gag aca caa atc cag aga
cgt cac cac acg gac att 3183Asn Tyr Gly Gly Glu Thr Gln Ile Gln Arg
Arg His His Thr Asp Ile 545 550
555ggt ttc atc atg gac aga ttt gtg aag atc caa agc ttg agc cca aca
3231Gly Phe Ile Met Asp Arg Phe Val Lys Ile Gln Ser Leu Ser Pro Thr
560 565 570cat gtc att gac ctc atg cag
gct cac caa cac ggt ctg gtg ggt gcc 3279His Val Ile Asp Leu Met Gln
Ala His Gln His Gly Leu Val Gly Ala 575 580
585ttg ctg cgt gca gcc acg tac tac ttt tct gac ctg gaa att gtt gta
3327Leu Leu Arg Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile Val Val590
595 600 605cgg cac gaa ggc
aat ctg acc tgg gtg ccc aac ggc gcc cct gaa tca 3375Arg His Glu Gly
Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ser 610
615 620gcc ctg ttg aac acc agc aac ccc act gcc
tac aac aag gca cca ttc 3423Ala Leu Leu Asn Thr Ser Asn Pro Thr Ala
Tyr Asn Lys Ala Pro Phe 625 630
635acg aga ctc gct ctc ccc tac act gcg ccg cac cgt gtg ctg gca aca
3471Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala Thr
640 645 650gtg tac aac ggg acg agt aag
tat gct gtg ggt ggt tca ggc aga aga 3519Val Tyr Asn Gly Thr Ser Lys
Tyr Ala Val Gly Gly Ser Gly Arg Arg 655 660
665ggc gac atg ggg tct ctc gcg gcg cga gtc gtg aaa cag ctt cct gct
3567Gly Asp Met Gly Ser Leu Ala Ala Arg Val Val Lys Gln Leu Pro Ala670
675 680 685tca ttt aac tac
ggt gca atc aag gcc gac gcc atc cac gaa ctt ctc 3615Ser Phe Asn Tyr
Gly Ala Ile Lys Ala Asp Ala Ile His Glu Leu Leu 690
695 700gtg cgc atg aaa cgg gcc gag ctc tac tgc
ccc aga ccg ctg ttg gca 3663Val Arg Met Lys Arg Ala Glu Leu Tyr Cys
Pro Arg Pro Leu Leu Ala 705 710
715ata gag gtg tct tcg caa gac agg cac aag caa aag atc att gca cca
3711Ile Glu Val Ser Ser Gln Asp Arg His Lys Gln Lys Ile Ile Ala Pro
720 725 730gca aag cag ctt ctg aat ttt
gac ctg ctc aag ttg gcc gga gac gtt 3759Ala Lys Gln Leu Leu Asn Phe
Asp Leu Leu Lys Leu Ala Gly Asp Val 735 740
745gag tcc aac ccc ggg cca ttc ttc ttt gct gac gtt agg tca aac ttt
3807Glu Ser Asn Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe750
755 760 765tca aag ttg gta
gac aca atc aac cag atg cag gag gac atg tcc aca 3855Ser Lys Leu Val
Asp Thr Ile Asn Gln Met Gln Glu Asp Met Ser Thr 770
775 780aaa cac ggg ccc gac ttc aac cgg ttg gtg
tcc gca ttt gag gaa ttg 3903Lys His Gly Pro Asp Phe Asn Arg Leu Val
Ser Ala Phe Glu Glu Leu 785 790
795gcc act gga gtt aaa gct atc agg acc ggt ctc gac gag gcc aaa ccc
3951Ala Thr Gly Val Lys Ala Ile Arg Thr Gly Leu Asp Glu Ala Lys Pro
800 805 810tgg tac aag ctt atc aaa ctc
cta agc cgc ctg tcg tgc atg gcc gct 3999Trp Tyr Lys Leu Ile Lys Leu
Leu Ser Arg Leu Ser Cys Met Ala Ala 815 820
825gtg gca gca cgg tcc aag gac cca gtc ctt gtg gcc atc atg ctg gcc
4047Val Ala Ala Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala830
835 840 845gac acc ggt ctc
gag cgt cag aga cct ctg aaa gtg aga gct aag ctc 4095Asp Thr Gly Leu
Glu Arg Gln Arg Pro Leu Lys Val Arg Ala Lys Leu 850
855 860cca cag cag gaa gga cct tac gct ggc ccg
ttg gag aga cag aaa ccg 4143Pro Gln Gln Glu Gly Pro Tyr Ala Gly Pro
Leu Glu Arg Gln Lys Pro 865 870
875ctg aaa gtg aaa gca aaa gcc ccg gtc gtc aag gaa gga cct tac gag
4191Leu Lys Val Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu
880 885 890gga ccg gtg aag aag cct gtc
gct ttg aaa gtg aaa gct aag aac ttg 4239Gly Pro Val Lys Lys Pro Val
Ala Leu Lys Val Lys Ala Lys Asn Leu 895 900
905ata gtc act gag agt ggt gcc cca ccg acc gac ttg caa aag atg gtc
4287Ile Val Thr Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met Val910
915 920 925atg ggc aac aca
aag cct gtt gag ctc atc ctt gac ggg aag aca gta 4335Met Gly Asn Thr
Lys Pro Val Glu Leu Ile Leu Asp Gly Lys Thr Val 930
935 940gcc atc tgt tgt gct act gga gtg ttt ggc
act gct tac ctc gtg cct 4383Ala Ile Cys Cys Ala Thr Gly Val Phe Gly
Thr Ala Tyr Leu Val Pro 945 950
955cgt cat ctt ttc gca gag aag tat gac aag atc atg ctg gat ggc aga
4431Arg His Leu Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg
960 965 970gcc atg aca gac agt gac tac
aga gtg ttt gag ttt gag att aaa gta 4479Ala Met Thr Asp Ser Asp Tyr
Arg Val Phe Glu Phe Glu Ile Lys Val 975 980
985aaa gga cag gac atg ctc tca gac gct gcg ctc atg gtg ctc cac cgt
4527Lys Gly Gln Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu His Arg990
995 1000 1005ggg aac cgc gtg
aga gat atc acg aaa cac ttt cgt gat aca gca aga 4575Gly Asn Arg Val
Arg Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg 1010
1015 1020atg aag aaa ggc acc ccc gtc gtc ggt gtg
gtc aac aac gcc gac gtt 4623Met Lys Lys Gly Thr Pro Val Val Gly Val
Val Asn Asn Ala Asp Val 1025 1030
1035ggg aga ctg att ttc tct ggt gag gcc ctc acc tac aag gat att gta
4671Gly Arg Leu Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val
1040 1045 1050gtg tgc atg gac gga gac acc
atg cct ggc ctc ttt gcc tac aaa gcc 4719Val Cys Met Asp Gly Asp Thr
Met Pro Gly Leu Phe Ala Tyr Lys Ala 1055 1060
1065gcc acc aag gca ggc tac tgt gga gga gcc gtt ctc gcc aag gac ggg
4767Ala Thr Lys Ala Gly Tyr Cys Gly Gly Ala Val Leu Ala Lys Asp
Gly1070 1075 1080 1085gcc gac
act ttc atc gtc ggc act cac tcc gca gga ggc aat gga gtt 4815Ala Asp
Thr Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val
1090 1095 1100gga tac tgc tca tgc gtt tcc
agg tcc atg ctt ctc aga atg aag gca 4863Gly Tyr Cys Ser Cys Val Ser
Arg Ser Met Leu Leu Arg Met Lys Ala 1105 1110
1115cac gtt gac cct gaa cca caa cac gag tagtaatttt
tctagaggat 4910His Val Asp Pro Glu Pro Gln His Glu 1120
1125ccctcgagtt tttattgact agttaatcat aagataaata atatacagca
ttgtaaccat 4970cgtcatccgt tatacgggga ataatattac catacagtat tattaaattt
tcttacgaag 5030aatatagatc ggtatttatc gttagtttat tttacattta ttaattaaac
atgtctacta 5090ttacctgtta tggaaatgac aaatttagtt atataattta tgataaaatt
aagataataa 5150taatgaaatc aaataattat gtaaatgcta ctagattatg tgaattacga
ggaagaaagt 5210ttacgaactg gaaaaaatta agtgaatcta aaatattagt cgataatgta
aaaaaaataa 5270atgataaaac taaccagtta aaaacggata tgattatata cgttaaggat
attgatcata 5330aaggaagaga tacttgcggt tactatgtac accaagatct ggtatcttct
atatcaaatt 5390ggatatctcc gttattcgcc gttaaggtaa ataaaattat taactattat
atatgtaatg 5450aatatgatat acgacttagc gaaatggaat ctgatatgac agaagtaata
gatgtagttg 5510ataaattagt aggaggatac aatgatgaaa tagcagaaat aatatatttg
tttaataaat 5570ttatagaaaa atatattgct aacatatcgt tatcaactga attatctagt
atattaaata 5630attttataaa ttttaataaa aaatacaata acgacataaa agatattaaa
tctttaattc 5690ttgatctgaa aaacacatct ataaaactag ataaaaagtt attcgataaa
gataataatg 5750aatcgaacga tgaaaaattg gaaacagaag ttgataagct aatttttttc
atctaaatag 5810tattatttta ttgaagtacg aagttttacg ttagataaat aataaaggtc
gatttttatt 5870ttgttaaata tcaaatatgt cattatctga taaagataca aaaacacacg
gtgattatca 5930accatctaac gaacagatat tacaaaaaat acgtcggact atggaaaacg
aagctgatag 5990cctcaataga agaagcatta aagaaattgt tgtagatgtt atgaagaatt
gggatcatcc 6050tctcaacgaa gaaatagata aagttctaaa ctggaaaaat gatacattaa
acgatttaga 6110tcatctaaat acagatgata atattaagga aatcatacaa tgtctgatta
gagaatttgc 6170gtttaaaaag atcaattcta ttatgtatag ttatgctatg gtaaaactca
attcagataa 6230cgaaacattg aaagataaaa ttaaggatta ttttatagaa actattctta
aagacaaacg 6290tggttataaa caaaagccat taccc
6315881126PRTFoot-and-mouth disease virus 88Met Gly Ala Gly
Gln Ser Ser Pro Ala Thr Gly Ser Gln Asn Gln Ser1 5
10 15Gly Asn Thr Gly Ser Ile Ile Asn Asn Tyr
Tyr Met Gln Gln Tyr Gln 20 25
30Asn Ser Met Asp Thr Gln Leu Gly Asp Asn Ala Ile Ser Gly Gly Ser
35 40 45Asn Glu Gly Ser Thr Asp Thr Thr
Ser Thr His Thr Thr Asn Thr Gln 50 55
60Asn Asn Asp Trp Phe Ser Lys Leu Ala Ser Ser Ala Phe Thr Gly Leu65
70 75 80Phe Gly Ala Leu Leu
Ala Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu 85
90 95Glu Asp Arg Ile Leu Thr Thr Arg Asn Gly His
Thr Thr Ser Thr Thr 100 105
110Gln Ser Ser Val Gly Val Thr His Gly Tyr Ser Thr Glu Glu Asp His
115 120 125Val Ala Gly Pro Asn Thr Ser
Gly Leu Glu Thr Arg Val Val Gln Ala 130 135
140Glu Arg Phe Tyr Lys Lys Tyr Leu Phe Asp Trp Thr Thr Asp Lys
Ala145 150 155 160Phe Gly
His Leu Glu Lys Leu Glu Leu Pro Ser Asp His His Gly Val
165 170 175Phe Gly His Leu Val Asp Ser
Tyr Ala Tyr Met Arg Asn Gly Trp Asp 180 185
190Val Glu Val Ser Ala Val Gly Asn Gln Phe Asn Gly Gly Cys
Leu Leu 195 200 205Val Ala Met Val
Pro Glu Trp Lys Glu Phe Asp Thr Arg Glu Lys Tyr 210
215 220Gln Leu Thr Leu Phe Pro His Gln Phe Ile Ser Pro
Arg Thr Asn Met225 230 235
240Thr Ala His Ile Thr Val Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln
245 250 255Tyr Lys Lys His Lys
Pro Trp Thr Leu Val Val Met Val Val Ser Pro 260
265 270Leu Thr Val Asn Asn Thr Ser Ala Ala Gln Ile Lys
Val Tyr Ala Asn 275 280 285Ile Ala
Pro Thr Tyr Val His Val Ala Gly Glu Leu Pro Ser Lys Glu 290
295 300Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr
Gly Gly Leu Val Thr305 310 315
320Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn Pro
325 330 335Pro Arg Thr Asn
Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp Val Ala 340
345 350Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp
Gly Lys Pro Tyr Val 355 360 365Thr
Thr Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu Ser 370
375 380Leu Ala Ala Lys His Met Ser Asn Thr Tyr
Leu Ser Gly Ile Ala Gln385 390 395
400Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe
Thr 405 410 415Gly Ser Thr
Asp Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro Pro 420
425 430Gly Val Glu Thr Pro Pro Asp Thr Pro Glu
Arg Ala Ala His Cys Ile 435 440
445His Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser Ile 450
455 460Pro Tyr Val Ser Ala Ala Asp Tyr
Ala Tyr Thr Ala Ser Asp Thr Ala465 470
475 480Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr
Gln Ile Thr His 485 490
495Gly Lys Ala Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly Lys
500 505 510Asp Phe Glu Leu Arg Leu
Pro Ile Asp Pro Arg Gln Gln Thr Thr Ala 515 520
525Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu Asn
Tyr Gly 530 535 540Gly Glu Thr Gln Ile
Gln Arg Arg His His Thr Asp Ile Gly Phe Ile545 550
555 560Met Asp Arg Phe Val Lys Ile Gln Ser Leu
Ser Pro Thr His Val Ile 565 570
575Asp Leu Met Gln Ala His Gln His Gly Leu Val Gly Ala Leu Leu Arg
580 585 590Ala Ala Thr Tyr Tyr
Phe Ser Asp Leu Glu Ile Val Val Arg His Glu 595
600 605Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu
Ser Ala Leu Leu 610 615 620Asn Thr Ser
Asn Pro Thr Ala Tyr Asn Lys Ala Pro Phe Thr Arg Leu625
630 635 640Ala Leu Pro Tyr Thr Ala Pro
His Arg Val Leu Ala Thr Val Tyr Asn 645
650 655Gly Thr Ser Lys Tyr Ala Val Gly Gly Ser Gly Arg
Arg Gly Asp Met 660 665 670Gly
Ser Leu Ala Ala Arg Val Val Lys Gln Leu Pro Ala Ser Phe Asn 675
680 685Tyr Gly Ala Ile Lys Ala Asp Ala Ile
His Glu Leu Leu Val Arg Met 690 695
700Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Ile Glu Val705
710 715 720Ser Ser Gln Asp
Arg His Lys Gln Lys Ile Ile Ala Pro Ala Lys Gln 725
730 735Leu Leu Asn Phe Asp Leu Leu Lys Leu Ala
Gly Asp Val Glu Ser Asn 740 745
750Pro Gly Pro Phe Phe Phe Ala Asp Val Arg Ser Asn Phe Ser Lys Leu
755 760 765Val Asp Thr Ile Asn Gln Met
Gln Glu Asp Met Ser Thr Lys His Gly 770 775
780Pro Asp Phe Asn Arg Leu Val Ser Ala Phe Glu Glu Leu Ala Thr
Gly785 790 795 800Val Lys
Ala Ile Arg Thr Gly Leu Asp Glu Ala Lys Pro Trp Tyr Lys
805 810 815Leu Ile Lys Leu Leu Ser Arg
Leu Ser Cys Met Ala Ala Val Ala Ala 820 825
830Arg Ser Lys Asp Pro Val Leu Val Ala Ile Met Leu Ala Asp
Thr Gly 835 840 845Leu Glu Arg Gln
Arg Pro Leu Lys Val Arg Ala Lys Leu Pro Gln Gln 850
855 860Glu Gly Pro Tyr Ala Gly Pro Leu Glu Arg Gln Lys
Pro Leu Lys Val865 870 875
880Lys Ala Lys Ala Pro Val Val Lys Glu Gly Pro Tyr Glu Gly Pro Val
885 890 895Lys Lys Pro Val Ala
Leu Lys Val Lys Ala Lys Asn Leu Ile Val Thr 900
905 910Glu Ser Gly Ala Pro Pro Thr Asp Leu Gln Lys Met
Val Met Gly Asn 915 920 925Thr Lys
Pro Val Glu Leu Ile Leu Asp Gly Lys Thr Val Ala Ile Cys 930
935 940Cys Ala Thr Gly Val Phe Gly Thr Ala Tyr Leu
Val Pro Arg His Leu945 950 955
960Phe Ala Glu Lys Tyr Asp Lys Ile Met Leu Asp Gly Arg Ala Met Thr
965 970 975Asp Ser Asp Tyr
Arg Val Phe Glu Phe Glu Ile Lys Val Lys Gly Gln 980
985 990Asp Met Leu Ser Asp Ala Ala Leu Met Val Leu
His Arg Gly Asn Arg 995 1000 1005Val
Arg Asp Ile Thr Lys His Phe Arg Asp Thr Ala Arg Met Lys Lys 1010
1015 1020Gly Thr Pro Val Val Gly Val Val Asn Asn
Ala Asp Val Gly Arg Leu1025 1030 1035
1040Ile Phe Ser Gly Glu Ala Leu Thr Tyr Lys Asp Ile Val Val Cys
Met 1045 1050 1055Asp Gly
Asp Thr Met Pro Gly Leu Phe Ala Tyr Lys Ala Ala Thr Lys 1060
1065 1070Ala Gly Tyr Cys Gly Gly Ala Val Leu
Ala Lys Asp Gly Ala Asp Thr 1075 1080
1085Phe Ile Val Gly Thr His Ser Ala Gly Gly Asn Gly Val Gly Tyr Cys
1090 1095 1100Ser Cys Val Ser Arg Ser Met
Leu Leu Arg Met Lys Ala His Val Asp1105 1110
1115 1120Pro Glu Pro Gln His Glu 1125
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