Live attenuated antigenically marked classical swine fever vaccine

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to the design of a double antigenically marked classical swine fever virus (CSFV) live attenuated candidate strain vaccine validated for absence of reversions. The FlagT4-mFT-Gv virus is a modified recombinant FlagT4 RB-C22 virus containing a multiple-mutated Flag insertion epitope in the E1 glycoprotein and a multiple-mutated WH303 epitope in the E2 glycoprotein.

2. Description of the Relevant Art

Classical swine fever (CSF) is a highly contagious disease of swine. The etiological agent, CSF virus (CSFV), is a small, enveloped virus with a positive, single-stranded RNA genome, classified as a member of the genus Pestivirus within the family Flaviridae (Becher et al. 2003. Virology 311: 96-104). The 12.5 kb CSFV genome contains a single open reading frame that encodes a 3898-amino-acid polyprotein and ultimately yields 11 to 12 final cleavage products (NH.sub.2-Npro-C-E.sup.ms-E1-E2-p7-N52-N53-NS4A-NS4B-NS5A-NS5B-COOH) through co- and post-translational processing of the polyprotein by cellular and viral proteases (Rice, C. M. 1996. In: Fundamental Virology, 3rd edition, Fields and Howley, eds., Lippincott Raven, Philadelphia, pp. 931-959). Structural components of the CSFV virion include the capsid (C) protein and glycoproteins E.sup.ms, E1, and E2. The E1 and E2 glycoproteins are anchored to the envelope at their carboxyl termini and E.sup.ms loosely associates with the viral envelope (Thiel et al. 1991. J. Virol. 65: 4705-4712; Weiland et al. 1990. J. Virol. 64: 3563-3569; Weiland et al. 1999. J. Gen. Virol. 80: 1157-1165). E1 and E2 are type I transmembrane proteins with an N-terminal ectodomain and a C-terminal hydrophobic anchor (Thiel et al., supra). E2 is the most immunogenic of the CSFV glycoproteins (Konig et al. 1995. J. Virol. 69: 6479-6486; van Gennip et al. 2000. Vaccine 19:447-459); Weiland et al. 1990, supra), inducing neutralizing antibodies, which provide protection against lethal CSFV challenge.

The two main policies used for CSFV control are prophylactic vaccination or non-vaccination with "stamping out" of exposed animals in the event of an outbreak. Countries considered free of CSF do not recommend the use of currently available live attenuated viruses as tools to control outbreaks of the disease, despite the proven efficacy of these vaccines in eliciting a rapid and solid protection against the virus (van Oirschot, J. T. 2003. Vet. Microbiol. 96: 367-384). The humoral immune response induced by these vaccines does not differ from that elicited by infections caused by wild-type viruses; hence, the use of CSFV live attenuated viruses has been hampered by their inability of inducing a response differentiable between infected and vaccinated animals, i.e., by their lack of DIVA capability. Thus, the use of a CSFV live attenuated virus with DIVA capabilities could significantly impact policies of disease control. CSFV subunit marker vaccines with DIVA capabilities have been developed using recombinant CSFV E2 envelope protein (Hulst et al. 1993. J. Virol. 67: 5435-5442; Van Rijn et al. 1996. J. Gen. Virol. 77: 2737-2745; Van Rijn et al. 1999. Vaccine 17: 433-440)). The onset of immunity elicited by subunit vaccines occurs two weeks post-vaccination, limiting their efficacy relative to traditional live attenuated virus vaccines when animals are exposed to CSFV shortly after vaccination (Bouma et al. 2000. Vaccine 18: 1374-1381; Uttenthal et al. 2001. Vet. Microbiol. 83: 85-106).

Recently, we reported the development of a CSFV experimental marker live attenuated virus strain, FlagT4v (Holinka et al. 2009. Virology 384:106-113). FlagT4v contains the synthetic epitope of Flag.RTM. (Sigma, St. Louis, Mo.) as an insertion within the 19 mer insertion of a previously modified E1. Flag.RTM. serves as a positive antigenic marker. FlagT4 also serves as a negative antigenic marker as the result of the abolition of a highly conserved CSFV-specific epitope recognized by monoclonal antibody WH303 (mAbWH303e) (Edwards et al. 1991. Vet Microbiol. 29:101-108). Immunization with FlagT4v induced complete protection against challenge with virulent CSFV Brescia, both at 3 and 28 days post-infection (DPI). Serological responses against both the Flag and mAbWH303 epitopes in animals immunized with FlagT4v allowed the discrimination of animals immunized with FlagT4v from animals challenged with CSFV Brescia.

FlagT4 was further analyzed as a candidate vaccine strain and its minimal protective dose, biosafety and attenuation stability were determined. Although FlagT4v was completely atoxic and showed a protective efficacy compatible with further development as a vaccine, it presented some degree of reversion to virulence when successively passed in swine. Thus, there was a need to modify the FlagT4 virus to obtain an attenuated virus that has the attributes of a positive and a negative marker and is also a successful live attenuated vaccine.

SUMMARY OF THE INVENTION

We have developed a novel classical swine fever mutant virus, the FlagT4-mFT-Gv virus.

In accordance with this discovery, it is an object of the invention to provide a recombinant classical swine fever virus (CSFV) mutant virus, the FlagT4-mFT-Gv virus, a modification of the antigenically-marked FlagT4 CSFV. The nucleotide sequence of FlagT4-mFT-Gv (SEQ ID NO:1) differs from the nucleotide sequence encoding the FlagT4 CSFV. While FlagT4-mFT-Gv (SEQ ID NO:1) encodes the same amino acids of the 20 mer Flag-containing insertion of the modified CSFV E1 glycoprotein of FlagT4, all the nucleotide triplets (codons) encoding said amino acids have been changed, if they could be changed because of the redundancy in codons encoding for the same amino acids, and thus differ from the nucleotide sequence (SEQ ID NO:3) encoding the region of the 20 mer Flag-containing insertion of the original FlagT4 live attenuated virus. Further, in the FlagT4-mFT-Gv, an additional change has been made in the T4 region of E2 where both the nucleotide sequence and the amino acid sequence of the T4 region of the modified CSFV E2 glycoprotein of the FlagT4 virus have been further modified. Thus, while the FlagT4-mFT-Gv, like FlagT4, lacks an immunodominant WH303 epitope in the modified T4 region of the E2 glycoprotein, the nucleotide and amino acid sequences of FlagT4 and FlagT4-mFT-Gv differ. The nucleotide sequence of T4 has been changed to encode a T4 region that differs from the T4 region of FlagT4 by one amino acid, namely, asparagine has been changed to glycine; and, further, those nucleotide triplets (codons) encoding the amino acids that remain the same in the T4 region of FlagT4-mFT-Gv and FlagT4 have also been changed where redundant codons for the same amino acids allow.

An added object of the invention is to provide immunogenic compositions comprising a viable recombinant classical swine fever virus mutant, FlagT4-mFT-Gv.

An additional object of the invention is to provide a rationally designed live attenuated CSFV effective to protect an animal from clinical CSF disease when challenged with virulent Brescia CSFV.

A further object of the invention is to provide a marker vaccine which can potentially distinguish between vaccinated animals and animals infected with CSFV.

A still further object of the invention is to provide a vaccine comprising at least one additional vaccine strain capable of inducing protection against CSF or against another porcine pathogen.

Another object of the invention is to provide a method for protecting an animal against CSF by administering an effective amount of rationally designed live attenuated CSFV vaccine.

An additional object of the invention is to provide a method for distinguishing animals infected with CSFV from animals vaccinated with said rationally designed live attenuated CSFV vaccine, comprising: analyzing serum to compare the serological responses to the marker sequences as determined by the ability of said animal serum to recognize the FLAG sequence and WH303 sequence antigens wherein the serologic profile observed for vaccinated animals can be differentiated from the serologic profile observed for wild-type infected animals.

Other objects and advantages of this invention will become readily apparent from the ensuing description.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the U.S. Patent and Trademark Office upon request and payment of the necessary fee.

FIGS. 1A and 1B are schematic representations depicting codon changes introduced into the Flag (FIG. 1A) and the T (FIG. 1B) areas of FlagT4-mFT-Gv as compared to the codon usage in the parental FlagT4v. In FIG. 1A, the nucleotide sequence showing changes in the triplet codons depicts nucleotides 2435-2488 of SEQ ID NO:1 and is identified in the Sequence Listing as SEQ ID NO:5. The nucleotide sequence with no changes in the triplet codons depicts nucleotides 2435-2488 of SEQ ID NO:3, the nucleotide sequence encoding the original FlagT4, and is identified in the Sequence Listing as SEQ ID NO:6. The amino acid sequence depicts amino acids 688 to 705 of SEQ ID NOs: 2 and 4 and is identified in the Sequence Listing as SEQ ID NO:7. Oligonucleotide primers used to introduce nucleotide substitutions are shown. The sequences of the Forward Primers (FP) and Reverse Primers (RP) of Flag 1, Flag 2, and Flag 3 are shown and are identified in the Sequence Listing by SEQ ID NOs: 8, 9, 10, 11, 12, and 13, respectively. FIG. 1B depicts the change of nucleotide sequence tca ttt aat atg gac (SEQ ID NO: 14) encoding SFNMD (SEQ ID NO:15) representing nucleotides 2921-2935 of SEQ ID NO:3 encoding amino acids 849-857 of SEQ ID NO: 4 (FlagT4) to the nucleotide sequence agt ttc gga atg gat (SEQ ID NO:16) encoding SFGMD (SEQ ID NO:17) representing nucleotides 2921-2935 of SEQ ID NO:1 encoding amino acids 849-857 of SEQ ID NO: 2 (FlagT4-mFt-Gv). The sequences of the Forward T4 Primer (FT4) and the Reverse T4 Primer (RT4) are shown and identified as SEQ ID NOs: 18 and 19, respectively.

FIG. 2 shows the antigenic profile of FlagT4-mFt-Gv. Sk6 cell cultures were infected with FlagT4-mFt-Gv and 4 days later fixed and stained by immunohistochemistry with mAbs WH303, anti-Flag or WH174.

DETAILED DESCRIPTION OF THE INVENTION

We have developed a live attenuated CSFV vaccine strain, FlagT4-mFT-Gv (SEQ ID NO:1), a modified version of the FlagT4v where changes in the codon usage were introduced into the genomic areas encoding for Flag and T4 to rectify the reversion of FlagT4 to the virulent genotype. The new virus, FlagT4-mFT-Gv, possesses the same amino acid sequence as FlagT4v except for one substitution, the substitution of asparagine by glycine at position 852 in the T4 region of the CSFV FlagT4 polypeptide. The nucleotide sequence of FlagT4-mFT-Gv (SEQ ID NO:1) differs from the nucleotide sequence encoding the FlagT4 CSFV. The nucleotide sequence of FlagT4-mFT-Gv (SEQ ID NO:1) encodes the same amino acids of the modified CSFV E1 glycoprotein of FlagT4; however, in FlagT4-mFT-Gv, all the nucleotide triplets (codons) of the modified CSFV E1 have been changed, if they could be changed because of the redundancy in codons encoding for the same amino acids, and thus differ from the nucleotide sequence of the original FlagT4 live attenuated virus in the E1 region. Further, in the FlagT4-mFT-Gv, both the nucleotide sequence and the amino acid sequence of the T4 region of the modified CSFV E2 glycoprotein of the FlagT4 virus have been further modified. The region covered by amino acid positions 849-857 of the WH303 epitope of E2 has been changed from TSFNMDTLR to TSFGMDTLR.

The nucleotide sequence (SEQ ID NO:1) of FlagT4-mFT-Gv differs from the nucleotide sequence (SEQ ID NO:3) encoding the FlagT4 CSFV. The nucleotide sequence of FlagT4-mFT-Gv (SEQ ID NO:1) encodes the polypeptide sequence (SEQ ID NO:2). The amino acid sequence of the 20 mer Flag-containing insertion of the modified CSFV E1 glycoprotein of FlagT4-mFT-Gv is the same as the amino acid sequence as is found in the 20 mer Flag-containing insertion of the modified CSFV E1 protein in FlagT4, i.e., amino acids 688-705 representing the Flag region of the E1 glycoprotein of SEQ ID NO:2 (FlagT4-mFT-Gv polypeptide) and amino acids 688-705 representing the E1 portion of SEQ ID NO:4 (FlagT4 polypeptide) are the same. However, all the nucleotide triplets of SEQ ID NO:1 (FlagT4-mFT-Gv) encoding those amino acids of the 20 mer Flag-containing insertion of the modified FlagT4-mFT-Gv E1 protein, i.e., those amino acids identical to the 20 mer Flag-containing insertion of FlagT4, have been changed and differ from the nucleotide triplets of SEQ ID NO:3. Thus, nucleotides 2435-2488 of SEQ ID NO:1 differ from nucleotides 2435-2488 of SEQ ID NO:3, but they encode the same amino acid sequences.

The T4 modification in the E2 glycoprotein of the original FlagT4 resulted in the E2 glycoprotein lacking an immunodominant WH303 epitope. Both the nucleotide sequence and the amino acid sequence of the T4 region of the modified CSFV E2 glycoprotein of FlagT4 have been further modified in the FlagT4-mFT-Gv. In FlagT4-mFT-Gv, the nucleotide sequence of T4 (nucleotide triplet 2927, 2928, 2929 of SEQ ID NO:1) has been changed to encode a T4 region (amino acid 852 of SEQ ID NO:2) that differs from the T4 region of FlagT4 (amino acid 852 of SEQ ID NO:4) by one amino acid, namely, asparagine has been changed to glycine; and, further, those nucleotide triplets (2921-2935 of SEQ ID NO:1) encoding the amino acids that remain the same in the T4 region of FlagT4-mFT-Gv and FlagT4 (amino acids 850, 851, 853 and 854) of SEQ ID NO:2 and SEQ ID NO:4, respectively, have also been changed.

FlagT4-mFT-Gv was shown to efficiently maintain its attenuated phenotype during the reversion to virulence assay. In addition, FlagT4-mFT-Gv was effective in inducing protection to swine against the challenge at 21 days post vaccination with Brescia virulent virus. A serological response against the Flag epitope in FlagT4-mFT-Gv-immunized animals allows the discrimination between a Flag/T4v-immunized and a Brescia-infected animal.

A vaccine is defined herein as a biological agent which is capable of providing a protective response in an animal to which the vaccine has been delivered and is incapable of causing severe disease. Administration of the vaccine results in immunity from a disease; the vaccine stimulates antibody production or cellular immunity against the pathogen causing the disease. Immunity is defined herein as the induction of a significant higher level of protection in a population of swine against mortality and clinical symptoms after vaccination compared to an unvaccinated group. In particular, the vaccine according to the invention protects a large proportion of vaccinated animals against the occurrence of clinical symptoms of the disease and mortality. The vaccine of the invention herein is a genetically engineered mutant virus vaccine. A marker vaccine is defined as a vaccine that, in conjunction with a diagnostic test, enables serological differentiation of vaccinated animals from infected animals. A mutation is understood to be a change in the genetic information of a "wild-type" or unmodified E1 or E2 gene of a parent CSFV strain which is able to express native E1 and E2 proteins. Thus, the E1 and E2 polypeptides expressed by the FlagT4-mFT-Gv mutant virus are changed: the E1 protein displays a Flag epitope within the altered (by insertion) E1 protein and the E2 protein lacks a wild-type immunodominant WH303 epitope. The FlagT4-mFT-Gv recombinant classical swine fever virus (CSFV) mutant comprising DNA encoding a mutation in CSFV E1 glycoprotein, wherein the mutation comprises a 20 mer insertion, wherein said 20 mer insertion comprises the FLAG.TM. epitope as a positive mutation marker in the mutant CSFV E1, wherein said recombinant CSFV mutant is a live attenuated CSFV and said FLAG.TM. epitope carried by said live attenuated CSFV is capable of being recognized by monoclonal antibodies specifically binding to said FLAG.TM. epitope, which serves as a positive marker for said attenuated CSFV mutant and further comprises another mutation in CSFV E2 glycoprotein in addition to said positive mutation marker in CSFV E1 glycoprotein, wherein the mutant CSFV E2 glycoprotein comprises a substitution mutation in the wild-type WH303 immunogenic epitope wherein amino acids are substituted with the epitope of TSFGMDTLR thus resulting in the elimination of the immunodominant WH303 epitope of the wild-type CSFV and wherein said recombinant CSFV mutant is live attenuated CSFV having both the positive marker as a result of the mutation in CSFV E1 glycoprotein and the negative marker as a result of the mutation in the wild-type WH303 epitope in CSFV E2 glycoprotein.

A vaccine against CSFV is provided that comprises a FlagT4-mFT-Gv virus mutant as defined above in a live form, and a pharmaceutically acceptable carrier or diluent. The vaccine according to the invention containing the live virus can be prepared and marketed in the form of a suspension or in a lyophilized form and additionally contains a pharmaceutically acceptable carrier or diluent customary used for such compositions. Carriers include stabilizers, preservatives and buffers. Suitable stabilizers are, for example SPGA (sucrose, phosphate, glutamate, and human. albumin), carbohydrates (such as sorbitol, mannitol, starch, sucrose, dextran, glutamate or glucose), proteins (such as dried milk serum, albumin or casein) or degradation products thereof. Suitable buffers are for example alkali metal phosphates. Suitable preservatives are thimerosal, merthiolate and gentamicin. Diluents include water, aqueous buffer (such as buffered saline), alcohols and polyols (such as glycerol).

If desired, the live vaccines according to the invention may contain an adjuvant. Examples of suitable compounds and compositions with adjuvant activity are well known in the art. Furthermore, nucleic acid sequences encoding polypeptides for pharmaceutical or diagnostic applications, in particular immunomodulators such as lymphokines, interferons or cytokines, may be incorporated into the vaccine.

A vaccine according to the invention can be prepared by conventional methods such as those commonly used for the commercially available live attenuated CSFV vaccines. Briefly, a susceptible substrate is inoculated with the FlagT4-mFT-Gv mutant and propagated until the virus has replicated to a desired titer after which FlagT4-mFT-Gv-containing material is harvested. Subsequently, the harvested material is formulated into a pharmaceutical preparation with immunizing properties.

Every substrate which is able to support the replication of FlagT4-mFT-Gv viruses can be used in the present invention, including Swine kidney cells (SK6) and primary cultures of swine peripheral blood macrophages.

The vaccine may be administered by intramuscular, intradermal, subcutaneous or intranasal inoculation or injection in an amount which is effective to protect the animal against challenge by a virulent strain of CSFV. This amount may vary according to the animal being inoculated, taking into consideration the size and weight of the animal. The vaccine according to the invention comprises an effective dosage of the FlagT4-mFT-Gv mutant as the active component, i.e. an amount of immunizing FlagT4-mFT-Gv material that will induce immunity in the vaccinated animals, swine, against challenge by a virulent CSFV. Immunity is defined herein as the induction of a significant higher level of protection in a population of swine against mortality and clinical symptoms after vaccination compared to an unvaccinated group. In particular, the vaccine according to the invention prevents a large proportion of vaccinated animals against the occurrence of clinical symptoms of the disease and mortality. Typically, the live vaccine can be administered in a dose of 10.sup.4-10.sup.5 TCID.sub.50. Effective amounts may be experimentally determined as necessary by those of skill in the art by following the guidance provided, for example, by Example 5.

In addition to the FlagT4-mFT-Gv mutant, the invention can also include combination vaccines comprising a vaccine strain capable of inducing protection against another porcine pathogen.

The FlagT4-mFT-Gv marker vaccine described above, in conjunction with a diagnostic method, has the potential of distinguishing between animals that are vaccinated with it and animals that are infected with naturally occurring CSFV strains or vaccinated with conventional CSFV vaccines.

The present invention also provides an invaluable tool to monitor CSFV control measures that may lead to eradication of CSFV if applied in large scale stamping out programs. This tool concerns a method for determining CSFV infection in swine comprising the step of examining a sample of the animal for the presence or absence of antibodies reactive with the immunodominant epitopes FLAG and WH303. The sample of the animal used in this method may be any sample in which CSFV or FlagT4-mFT-Gv antibodies can be detected, e.g. a blood, serum or tissue sample.

The design of the immunoassay may vary. For example, the immunoassay may be based upon competition or direct reaction. Furthermore, protocols may use solid supports or may use cellular material. The detection of the antibody-antigen complex may involve the use of labeled antibodies; the labels may be, for example, enzymes, fluorescent, chemiluminescent, radioactive or dye molecules.

Suitable methods for the detection of WH303 and FLAG in the sample include, for example, the enzyme-linked immunosorbent assay (ELISA), immunofluorescent tests and Western blot analysis.

Porcine anti-CSFV sera raised against the FlagT4-mFT-Gv mutant according to the present invention has the potential of being distinguishable from porcine sera raised against naturally occurring CSFV strains and conventional CSFV vaccine strains. Thus, the FlagT4-mFT-Gv mutant has the potential of being a marker vaccine.

Thus, the particular CSFV mutant, FlagT4-mFT-Gv, according to the present invention has the potential of producing antiserum in pigs wherein the antisera are lacking in antibodies that react with an immunodominant epitope, WH303. Such antiserum would score negative in a direct- or blocking WH303 enzyme-linked immunosorbant assay (ELISA).

In an ELISA to detect porcine anti-FLAG and anti-WH303 epitope porcine antibodies, microtitration plates are coated with FLAG-bearing E1 protein (or a FLAG antigen) or wild type E2 protein bearing the WH303 epitope (or an E2 fragment bearing the WH303 epitope). Next, the wells of the coated plates are filled with porcine serum and serial dilutions are made. After incubation, porcine anti-FLAG or anti-WH303 epitope protein serum antibodies are determined by detecting antibody (monoclonal or polyclonal) with the same specificity as the coated one, but which is labeled (e.g. with biotin). The labeled antibody will occupy the free antigens that have not been occupied by anti-WH303 epitope or anti-FLAG antibodies in the porcine serum. For example, horse radish peroxidase coupled to avidin may be added and the amount of peroxidase is measured by an enzymatic reaction. If no antibodies against FLAG-marked E1 or the E2 WH303 epitope are present in the porcine serum sample then a maximum absorption is obtained. If the serum contains many antibodies against the WH303 epitope then a low absorption is expected. Alternatively, after the incubation with porcine serum, the amount of antibodies present in the serum that bound to the WH303 epitope antigen may be determined directly by using an anti-porcine conjugate followed by the enzymatic reaction.

In a sandwich ELISA the wells of a polystyrene microtitration plate can be coated with a monoclonal antibody directed against the WH303 epitope protein, i.e., mAb WH303. Next, the wells of these coated plates are incubated with antigen. After the antigen is captured, the wells are filled with the porcine serum and serial dilutions are made. Subsequently, the protocol as described above may be followed.

In another diagnostic test (Western blot analysis), the FLAG-marked E1 polypeptide or FLAG fragment or the WH303 epitope-containing material is subjected to SDS-PAGE. Next, the separated proteins are electroblotted onto nitro-cellulose membrane. Thereafter, the membranes can be cut into lanes and the lanes are incubated with the porcine serum. The presence in the sample of antibodies specific for the WH303 epitope or antibodies specific for FLAG can be determined by examining whether antibodies bound to the WH303 epitope or the FLAG antigen, for example by using an anti-porcine conjugate followed by an enzymatic reaction. If antibodies against the WH303 epitope or FLAG are present then a band of the appropriate size is identifiable.

The WH303 epitope-containing polypeptide may be any WH303 epitope-containing protein or fragment comprising material which allows the formation of the WH303 epitope antigen-WH303 mAb complex. Similarly, the FLAG-containing polypeptide may be any FLAG-containing protein or fragment comprising material which allows the formation of the FLAG (antigen)-anti-FLAG (antibody) complex. Preferably, the WH303 epitope antigen and the FLAG antigen comprise the expression product of a conventional recombinant host cell or virus, e.g. such as E. coli expressed or baculovirus expressed protein. In a further embodiment of the present invention, a diagnostic test kit is provided which is suitable for performing the diagnostic test according to the invention as described above.

In particular, a diagnostic test kit is provided which comprises in addition to the components usually present, the WH303 epitope antigen or FLAG antigen (if desired, coated onto a solid phase) as the immunological reagent. Other components usually present in such a test kit include, biotin or horseradish peroxidase conjugated antibodies, enzyme substrate, washing buffer etc.

To determine CSFV (BICv) WH303 antigen or FLAG antigen in a test sample from an animal in the field, WH303 mAb and anti-FLAG mAb are used as the immunological reagent, preferably fixed to a solid phase. The test sample is added, and after an incubation time allowing formation of the antibody-antigen complex, a second labeled antibody may be added to detect the complex.

Typically, the absorbance (OD) cut-off value for the ELISA to differentiate positive from negative samples is set at three standard deviations above the average P/N ratios of negative control samples from pigs (where P=the OD of samples from wells coated with a relevant peptide coupled to a carrier molecule and; N=the OD of samples from wells coated with the carrier molecule). A carrier molecule can be a carrier protein, such as BSA, ovalbumin, KLH, a carbohydrate chain or a synthetic amino acid chain.

In an alternative embodiment of the diagnostic method the presence of specific antibodies in porcine serum is examined by incubating the serum and an appropriate antigen in the presence of a monoclonal antibody that specifically reacts with an epitope located within the E2 region.

The recombinant FlagT4-mFT-Gv mutant according to the invention is obtained after transfection of suitable cells (e.g., SK6 cells) with the synthetic RNA transcript of the FlagT4-mFT-Gv mutant genome by electroporation. Finally the recombinant FlagT4-mFT-Gv mutant is harvested from the supernatant of the transformed cells.

It has also been found that FlagT4-mFT-Gv mutant according to the present invention is able to induce a protective immune response, i.e. animals immunized with a vaccine comprising the FlagT4-mFT-Gv mutant are protected against virulent challenge. Moreover, it has been found that antisera of animals infected with naturally occurring CSFV comprise antibodies directed to the WH303 epitope of BICv. Antisera from animals infected with the FlagT4-mFT-Gv mutant virus according to the present invention can be tested according to the methods described above to determine their reactivity with the Flag and WH303 epitope. In addition, it has been found that the FlagT4-mFT-Gv mutant virus as described above is attenuated if compared with the parent BICv virus which is able to produce the native E1 and E2 proteins.

EXAMPLES

Having now generally described this invention, the same will be better understood by reference to certain specific examples, which are included herein only to further illustrate the invention and are not intended to limit the scope of the invention as defined by the claims.

Example 1

Viruses and Cell Cultures

Swine kidney cells (SK6) (Terpstra et al. 1990. Dtsch. Tierarztl. Wochenschr. 97: 77-79), free of BVDV, were cultured in Dulbecco' minimal essential medium (DMEM) (Gibco, Grand Island, N.Y.) with 10% fetal calf serum (FCS) (Atlas Biologicals, Fort Collins, Colo.). CSFV derived from full-length cDNA copies, including CSFV Brescia strain (BICv) (Risatti et al. 2005a. J. Virol. 79: 3787-3796), RB-C22v (Risatti et al. 2005b. Virology 343: 116-127), T4v (Risatti et al. 2006. Virology 355: 94-101), and FlagT4v (Holinka et al., supra) were propagated in SK6 cells. Titration of CSFV from clinical samples was performed using SK6 cells in 96 well plates (Costar, Cambridge, Mass.). Viral infectivity was detected, after 4 days in culture, by immunoperoxidase assay using the CSFV monoclonal antibody (mAb) WH174 (kindly provided by Georgina Ibata, Veterinary Laboratory Agency, UK) or mAb WH303 (Edwards et al., supra), and the Vectastain ABC kit (Vector Laboratories, Buringames, Calif.). MAb WH174 recognizes and binds to the E2 protein of CSFV at an epitope different from that recognized by mAb WH303. Titers were calculated using the method of Reed and Muench (1938. Amer. J. Hygiene 27: 493-497) and expressed as TCID.sub.50/ml. As performed, test sensitivity was >1.8 TCID.sub.50/ml.

Example 2

DNA Sequencing and Analysis

Full-length infectious clones and in vitro rescued viruses were completely sequenced with CSFV-specific primers by the dideoxynucleotide chain-termination method (Sanger et al. 1977. Proc. Natl. Acad. Sci. USA 74: 5463-5467). Sequencing reactions were prepared with the Dye Terminator Cycle Sequencing Kit (Applied Biosystems, Foster City, Calif.). Reaction products were sequenced on a PRISM 3730xl automated DNA Sequencer (Applied Biosystems). Sequence data were assembled with the Phrap software program (Retrieved from the Internet: phrap.org), with confirmatory assemblies performed using CAP3 (Huang and Madan. 1999. Genome Res. 9: 868-877). The final DNA consensus sequence represented, on average, five-fold redundancy at each base position. Sequence comparisons were conducted using BioEdit software (Retrieved from the Internet: mbio.ncsu.edu/BioEdit/bioedit.html).

Example 3

Development of FlagT4-mFT-Gv

Infectious clone (IC) encoding for the FlagT4-mFT-Gv was designed and developed by modification of the FlagT4v infectious clone. Basically, the Flag area, i.e., the Flag/modified 20 mer insertion of FlagT4v infectious clone, was modified in three different progressive steps (Blocks 1, 2, 3 of FIG. 1A) by site-directed mutagenesis using the primers depicted in FIG. 1A. The site-directed mutagenesis resulted in the new mutant virus FlagT4-mFT-Gv having the identical amino acid sequence as the parent FlagT4v infectious clone virus; however, the codons used to encode the identical amino acids of the mutant virus FlagT4-mFT-Gv were changed (mutated) as compared to the parental virus (FlagT4) as shown in FIG. 1A. Similarly, changes in codon usage were also introduced in the T4 area by a single site-directed mutagenesis step as shown in FIG. 1B. In addition, an amino acid substitution was also included, that is, Asparagine was replaced by Glycine, at position 852 of the CSFV polypeptide (FIG. 1B). Thus, the original FlagT4 peptide SFNMD was replaced by SFGMD where the amino acid asparagine (N) was replaced by glycine (G) and while the remaining amino acids are the same as those of the FlagT4 parent, the codons encoding three of the four remaining (unchanged) amino acids have been changed by site-directed mutagenesis. Altogether, in the new FlagT4-mFT-Gv, one amino acid and 28 nucleotides have been changed.

Infectious RNA was in vitro transcribed from full-length infectious clones of the FlagT4-mFT-Gv and used to transfect SK6 cells as described earlier (Risatti et al., 2005a). Viruses were rescued from transfected cells by day 4 post-transfection. Full-length nucleotide sequences of the rescued virus genomes were identical to parental DNA plasmids, confirming that only predicted mutations were reflected in rescued viruses.

Stability of the attenuated phenotype of FlagT4-mFT-Gv was assessed by the reversion to virulence protocol. Basically, the protocol follows OIE regulations with minor modifications. In brief, a group of five animals (5-6 week old commercial female pigs) were inoculated intramuscularly (IM) with 10.sup.5 of FlagT4-mFT-Gv. Animals were kept under observation for 7 days and clinical signs and body temperature were recorded. At day 7.sup.th pi, animals were euthanized and their palatine tonsils removed. Presence of virus was detected in tonsils by virus isolation and by immunohistochemistry using mAb anti-Flag (Sigma) for virus detection. Tonsils were then macerated in a 10% w/v suspension and a pool of tonsil tissue suspension was used to inoculate a new group of 5 animals. This procedure was repeated 5 times until no virus was detected in tonsils of at least one of the five animals evaluated.

There was no presence of CSF-related symptoms in any of the animals in any of the groups. Virus was isolated from tonsils macerates only after the first passage at very low titer (2.8.times.10.sup.5 TCID.sub.50). These results indicate that FlagT4-mFT-Gv was stably attenuated in swine.

Example 4

Antigenic Profile of FlagT4-mFT-Gv

Infectious RNA was in vitro transcribed from full-length infectious clones of the FlagT4-mFT-Gv and used to transfect SK6 cells as described earlier (Risatti et al. 2005a, supra). Viruses were rescued from transfected cells by day 4 post-transfection.

SK6 cell cultures were infected with FlagT4-mFT-Gv and 4 days later fixed and stained by immunohistochemistry with mAbs WH303, anti-Flag or WH174 in order to analyze its antigenic phenotype. FlagT4-mFT-Gv strongly reacts with mAbs anti-Flag and WH174 and completely fails to react with mAb WH303. Conversely, BICv reacts with mAbs WH303 and WH174, but failed to react with mAb anti-Flag (FIG. 2).

Example 5

Animal Studies

For protection studies, female commercial pigs (40-50 lbs) were allocated in two groups harboring 5 animals each and immunized intramuscularly (IM) with one ml containing 10.sup.5 TCID.sub.50 of FlagT4-mFT-Gv or mock vaccinated. At 21 days post immunization (pi), animals were intranasally (IN) challenged with 10.sup.5 TCID.sub.50 BICv. Clinical signs and body temperature were recorded daily throughout the experiment as previously described (Risatti et al., 2005b). Blood, serum, and nasal swabs were collected at 7, 14 and 21 days pi and 4, 7, 11, 15, and 21 days post-challenge, with blood obtained from the anterior vena cava in EDTA-containing tubes (Vacutainer) for total and differential white blood cell counts. Total and differential white blood cell and platelet counts were obtained using a Beckman Coulter ACT (Beckman Fullerton, Coulter, Calif.). Tissue samples of palatine tonsil were collected from animals at death or during postmortem examination at 21 days pc and processed as described before (Risatti et al. 2005b, supra) to detect the presence of both FlagT4v and BICv.

The ability of FlagT4-mFT-Gv to induce protection against virulent BICv was assessed in late vaccination-exposure experiments. Groups of pigs (n=6) were IM-inoculated with FlagT4-mFT-Gv and challenged at 21 DPI. Mock-vaccinated control pigs receiving BICv only (n=6) developed anorexia, depression, and fever by 4 days post-challenge (DPC), a marked reduction of circulating leukocytes and platelets by 4 DPC (data not shown), and died or were euthanized in extremis by 6-9 DPC. Pigs challenged at 21 days post-FlagT4-mFT-Gv infection were protected, remaining clinically normal, with no alterations of hematological profiles (data not shown) or presence of fever (Table 1).

TABLE-US-00001 TABLE 1 Survival and fever response in FlagT4-mFT-Gv-vaccinated animals following challenge with BICv. Fever Group No. of Mean time Maximum Vaccinated Survivors/ to Death Days to Onset Duration Daily Temp with Total (Days .+-. SD) (Days .+-. SD) (Days .+-. SD) (.+-.SD) Mock 0/6 7.5 (1.5) 4.5 (0.57) 4.5 (1.72) 105.6 (.32) FlagT4-mFT-Gv 6/6 -- -- -- 102.4 (.24)

Viremia of vaccinated and challenged exposed animals was examined at different times post challenge (Table 2). Detection was performed using mAb WH303, which reacts specifically with the challenge virus BICv. As expected, in mock-vaccinated control animals, viremia was observed within 4 days after challenge, with virus titers remaining high by 7 DPC, the last time point tested before animals died or were euthanized. Conversely, animals inoculated with FlagT4-mFT-Gv and challenged with BICv at 21 DPI did not present viremia (Table 2).

TABLE-US-00002 TABLE 2 Viremia in FlagT4-mFT-Gv-infected animals after challenge with virulent BICv. Viremia Group Vaccinated With Duration (Days .+-. SD) Maximum (.+-.SD) Mock 7.75 (0.95) 7.1 (0.25) FlagT4-mFT-Gv 0 Neg

These results indicate that protection induced by FlagT4-mFT-Gv was complete, preventing both the presentation of CSF-related clinical signs and the replication of the challenge virus.

As a summary, we present here an approach for rationally developing of an experimental live attenuated marker CSFV vaccine strain, FlagT4-mFT-Gv, which is a derivative of the previous developed FlagT4v. Both FlagT4v and FlagT4-mFT-Gv harbor functional positive and negative antigenic markers that confer potential DIVA capabilities. The virus elicits solid protection against challenge with highly virulent BICv (Brescia strain) by 21 DPI when administered intramuscularly in swine. Potentially, response to FlagT4-mFT-Gv can be distinguished from a wild-type CSF virus (i.e., Brescia strain) by means of serology.

Attenuation of the parental FlagT4v was achieved by manipulating two independent novel genetic determinants of viral virulence (Risatti et al., 2005b; 2006) (i) an insertion of 21 codons in the genomic area encoding for structural glycoprotein E1 and (ii) a change in 5 codons encoding for a the stretch of amino residues in structural glycoprotein E2. FlagT4-mFT-Gv encodes the same amino acids in E1 as are found in FlagT4 E1 but a change in the nucleotide codon produces a substitution of Asparagine by Glycine at position 852. The nucleotide codons encoding E1 and E2 have been changed; thus, the nucleotide sequence of FlagT4-mFT-Gv differs from FlagT4 in the insertion of E1 and the T4 region of E2. SEQ ID NO:1 differs from SEQ ID NO:3. Results presented here demonstrated that these genomic changes produce a stabilization of the attenuated phenotype in FlagT4-mFT-Gv in comparison with the unstable one of the parental FlagT4v.

As a summary, results presented here demonstrated that genetic changes introduced in FlagT4-mFT-Gv produce a stabilization of its attenuated phenotype and that FlagT4-mFT-Gv induce a solid protection against the challenge in animals vaccinated 21 days earlier.

All publications and patents mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference.

The foregoing description and certain representative embodiments and details of the invention have been presented for purposes of illustration and description of the invention. It is not intended to be exhaustive or to limit the invention to the precise forms disclosed. It will be apparent to practitioners skilled in this art that modifications and variations may be made therein without departing from the scope of the invention.

SEQUENCE LISTINGS

1

19112357DNAClassical Swine Fever Virus 1gtatacgagg ttagttcatt ctcgtgtaca tgattggaca aatcaaaatc tcaatttggt 60tcagggcctc cctccagcga cggccgagct gggctagcca tgcccacagt aggactagca 120aacggaggga ctagccgtag tggcgagctc cctgggtggt ctaagtcctg agtacaggac 180agtcgtcagt agttcgacgt gagcagaagc ccacctcgag atgctatgtg gacgagggca 240tgcccaagac acaccttaac cctagcgggg gtcgttaggg tgaaatcaca ccatgtgatg 300ggagtacgac ctgatagggt gctgcagagg cccactatta ggctagtata aaaatctctg 360ctgtacatgg cacatggagt tgaatcattt tgaactttta tacaaaacaa acaaacaaaa 420accaatggga gtggaggaac cggtatacga tgtaacgggg agaccattgt ttggagaccc 480aagtgaggta cacccacaat caacattgaa gctaccacat gataggggga gaggcaacat 540caaaacaaca ctgaagaacc tacctaggag aggtgactgc aggagtggca accacctagg 600cccggttagt gggatatatg taaagcccgg ccctgtcttt tatcaggact acatgggccc 660agtctatcat agagcccctc tagagttttt tgacgaagca cagttttgtg aggtgaccaa 720aaggataggt agggtgacag gtagtgacgg aaagctttac catatatacg tgtgcatcga 780tggttgcatc ctgctgaagc tagccaagag gggcgagcca agaaccctga agtggattag 840aaatctcacc gactgtccat tgtgggttac cagttgttct gatgatggtg caagtgcaag 900taaagagaag aaaccagata ggatcaacaa gggtaaatta aagatagccc caaaagagca 960tgagaaggac agcaggacta agccacctga tgctacgatt gtagtggaag gagtaaaata 1020ccaggtcaaa aagaaaggta aagttaaggg aaagaatacc caagacggcc tgtaccacaa 1080caagaataaa ccaccagaat ctaggaagaa attagaaaaa gccctattgg catgggcagt 1140gatagcaatt atgttatacc aacctgttgc agccgaaaat ataactcaat ggaacctgag 1200tgacaacggt accaatggta tccagcacgc tatgtacctt agaggagtca gcagaagctt 1260gcatgggatc tggccagaaa aaatatgcaa aggagtcccc acctacctgg ccacagacac 1320ggaactgaga gaaatacagg gaatgatgga tgccagcgag gggacaaact atacgtgctg 1380taagttacag agacatgaat ggaacaaaca tggatggtgt aactggtata acatagaccc 1440ctggatacag ttgatgaata gaacccaagc aaacttggca gaaggccctc cgagcaagga 1500gtgcgccgtg acttgcaggt acgataaaaa tgctgacatt aacgtggtca cccaggccag 1560aaacaggcca accaccctaa ctggctgcaa gaaagggaaa aatttttctt ttgcgggtac 1620agttatagag ggcccatgta atttcaacgt ttctgttgag gatatcttat atggggatca 1680tgagtgtggc agtctactcc aggatacggc tctataccta gtagatggaa tgaccaacac 1740tatagagaga gccaggcagg gagccgcgag ggtgacatct tggctaggga ggcaactcag 1800aactgccggg aagaggttgg agggcagaag caaaacctgg tttggtgcct atgccctatc 1860accttattgt aatgtgacaa gcaaaatagg gtacatatgg tacactaaca actgtacccc 1920ggcttgcctc cccaaaaata caaagataat aggccccggt aaatttgaca ctaacgcgga 1980agacggaaag attctccatg agatgggggg ccacctatca gaatttctgc tgctctctct 2040ggtcgttctg tctgacttcg cccctgaaac agccagcgcg ttatacctca ttttgcacta 2100cgtgatccct caatcccatg aagaacctga aggctgtgac acaaaccagc tgaatttaac 2160agtggaactc aggactgaag acgtgatacc atcatcagtc tggaatgttg gcaaatatgt 2220gtgtgttaga ccagactggt ggccatatga aaccaaggtg gctttgttat ttgaagaggc 2280aggacaggtc gtaaagttag ccttgcgggc actgagggat ttaaccaggg tctggaatag 2340cgcatcaacc acggcattcc tcatctgctt gataaaagta ttaagaggac aggtcgtgca 2400aggtgtgata tggctgttac tggtaactgg ggcaccagtg agttgcaccc acttagccgc 2460ggccgactat aaagacgatg atgacaaagg ggcacaaggc cggctagcct gcaaggaaga 2520tcacaggtac gctatatcaa caaccaatga gatagggcta cttggggccg aaggtctcac 2580taccacctgg aaagaataca accacaattt gcaactggat gatgggaccg tcaaggccat 2640ctgcatggca ggttccttta aagtcacagc acttaatgtg gttagtagga ggtatctggc 2700atcattacat aaggacgctt tacccacttc cgtgacattc gagctcctgt tcgacgggac 2760cagcccattg accgaggaaa tgggagatga cttcgggttc ggactgtgtc cgtatgatac 2820gagccctgta gtcaagggaa agtacaacac aaccttgttg aatggtagtg cattctacct 2880agtttgccca atagggtgga cgggtgttat agagtgcacg agtttcggaa tggatactct 2940gagaacagaa gtggtaaaga ccttcagaag agagaaaccc tttccgtaca gaagggattg 3000tgtgaccact acagtggaaa atgaagatct attctactgt aaatgggggg gcaattggac 3060atgtgtgaaa ggtgaaccag tgacctacac gggggggcca gtaaaacaat gcagatggtg 3120tggcttcgac ttcaatgagc ctgacggact cccacactac cccataggta agtgcatttt 3180ggcaaatgag acaggttaca gaatagtgga ttcaacggac tgtaacagag atggcgttgt 3240aatcagcaca gaggggagtc atgagtgctt gattggtaac acaactgtca aggtgcatgc 3300attagatgaa agactaggcc ctatgccatg caggcctaag gagatcgtct ctagtgcggg 3360acctgtaagg aaaacttcct gtacattcaa ctacgcaaaa actctgagga acaggtatta 3420tgagcccagg gacagctatt tccaacaata tatgctcaag ggcgagtatc agtactggtt 3480tgatctggat gtgaccgacc gccactcaga ttacttcgca gaattcattg tcttggtggt 3540ggtggcactg ttgggaggaa gatatgtcct gtggctaata gtgacctaca tagttctaac 3600agaacaactc gccgctggtc tacagttagg ccagggtgag gtagtgttaa tagggaactt 3660aatcacccac acagatattg aggttgtagt atatttctta ctgctctatt tggtcatgag 3720agatgagcct ataaagaaat ggatactact gctgttccat gctatgacca acaatccagt 3780taagaccata acagtggcac tgctcatggt tagcggggtt gccaagggtg gaaagataga 3840tggtggttgg cagcggctgc cggagaccaa ctttgatatc caactcgcgc tgacagttat 3900agtagtcgct gtgatgttgc tggcaaagaa agatccgact accgtcccct tggttataac 3960ggtggcaacc ctgagaacgg ctaagataac taatggactt agtacagatc tagccatagc 4020tacagtgtca acagctttgc taacctggac ctacattagt gactattata aatacaagac 4080cttgctacag taccttatta gcacagtgac aggtatcttc ttgataaggg tactgaaggg 4140ggtaggtgag ttagatttac acaccccaac cttaccatct tacagacccc tcttcttcat 4200cctcgtgtac ctcatttcca ctgcagtggt aacaagatgg aatctggaca tagccggatt 4260gctgctgcag tgtgtcccaa cccttttaat ggttttcacg atgtgggcag acatccttac 4320cctgatcctc atactgccta cttacgagtt gacaaaacta tattacctca aggaagtgaa 4380gattggggca gaaaggggct ggttgtggaa gaccaacttc aagagggtaa atgacatata 4440cgaagttgac caagctggtg agggggtgta ccttttccca tcaaaacaaa agacaggtac 4500aataacaggt actatgttgc cattgatcaa agccatactc ataagttgca tcagcaataa 4560gtggcaattt atatatctat tgtacttgat attcgaagtg tcttactacc ttcacaagaa 4620gatcatagat gaaatagcag gagggaccaa cttcatctcg agacttgtag ccgctctgat 4680tgaagccaat tgggcctttg acaacgaaga agttagaggt ttaaagaagt tcttcctgct 4740gtctagtagg gttaaagaac tgatcatcaa acacaaagtg aggaatgaag tgatggtcca 4800ctggtttggc gacgaagagg tctatgggat gccgaagctg gttggcttag tcaaggcagc 4860aacactgagt aaaaataaac attgtatttt gtgcaccgtc tgtgaaaaca gagagtggag 4920aggagaaacc tgcccaaaat gcggccgttt tgggccacca gtgacctgtg gcatgaccct 4980agccgacttt gaagaaaaac actataagag gattttcttt agagaggatc aatcagaagg 5040gccggttagg gaggagtatg cagggtatct gcaatataga gccagagggc aattattcct 5100gaggaatctc ccggtgctag caacaaaagt caagatgctc ctggtcggaa atcttgggac 5160ggaggtgggg gatttggaac accttggctg ggtgctcaga gggcctgccg tttgcaagaa 5220ggttaccgaa catgagaaat gcaccacatc cataatggac aaattaactg ctttcttcgg 5280tgttatgcca aggggcacca cacctagagc ccctgtgaga ttccccacct ctctcttaaa 5340gataagaagg gggctggaaa ctggctgggc gtacacacac caaggtggca tcagttcagt 5400ggaccatgtc acttgtggga aagacttact ggtatgtgac actatgggcc ggacaagggt 5460tgtttgccaa tcaaataaca agatgacaga cgagtccgag tatggagtta aaactgactc 5520cggatgcccg gagggagcta ggtgttacgt gttcaaccca gaggcagtta acatatccgg 5580gactaaagga gccatggtcc acttacaaaa aactggagga gaattcacct gtgtgacagc 5640atcagggact ccggccttct ttgatctcaa gaacctcaaa ggctggtcag ggctgccgat 5700atttgaggca tcaagtggaa gagtagtcgg cagggttaag gtcgggaaga atgaggactc 5760taaaccaacc aagcttatga gtggaataca aacagtctcc aaaagtacca cagacttgac 5820agaaatggta aagaaaataa caaccatgaa caggggagaa ttcagacaaa taacccttgc 5880cacaggtgcc ggaaaaacca cggaactccc tagatcagtc atagaagaga taggaaggca 5940taagagggtc ttggtcttga tccctctgag ggcggcagca gagtcagtat accaatatat 6000gagacaaaaa cacccaagca tagcattcaa cttgaggata ggggagatga aggaagggga 6060catggccaca gggataacct atgcctcata tggttacttc tgtcagatgc cacaacctaa 6120gctgcgagcc gcgatggttg agtactcctt catattcctt gatgagtacc actgtgccac 6180ccccgaacaa ttggctatca tgggaaagat ccacagattt tcagagaacc tgcgggtagt 6240agccatgacc gcaacaccag caggcacggt aacaactaca gggcaaaaac accctataga 6300agaatacata gccccagaag tgatgaaggg ggaagactta ggttcagagt acttggacat 6360agctggacta aagataccag tagaggagat gaagagtaac atgctggtct ttgtgcccac 6420aaggaacatg gctgtagaga cggcaaagaa actgaaagct aagggttata actcaggcta 6480ctattatagt ggagaggatc catctaacct gagggtggta acatcacagt ccccgtacgt 6540ggtggtagca accaacgcaa tagaatcagg tgttactctc ccagacttgg atgtggtcgt 6600cgacacaggg cttaagtgtg aaaagaggat acggctgtca cctaagatgc ccttcatagt 6660gacgggcctg aagagaatgg ctgtcacgat tggggaacaa gcccagagaa gggggagagt 6720tgggagagtg aagcctggga gatactacag gagtcaagaa acccccgttg gttccaaaga 6780ttaccattac gacctactgc aagcacagag gtacggtata gaagatggga taaacatcac 6840caaatctttt agagagatga attatgattg gagcctttat gaggaggata gtctgatgat 6900tacacaattg gaaatcctca acaatctgtt gatatcagaa gagctaccaa tggcagtaaa 6960aaatataatg gccaggactg accacccaga accaatccaa ctggcgtaca acagctacga 7020aacgcaggtg ccagtgctat tcccaaaaat aaaaaatgga gaggtgactg acagttacga 7080taactatacc ttcctcaacg caagaaagct gggggatgat gtacctccct acgtgtatgc 7140cacagaggat gaggacttag cggtagagct gctgggctta gactggccgg accctgggaa 7200ccaaggaacc gtggaggctg gtagagcact aaaacaagta gttggtctat caacagctga 7260gaacgccctg ttagtagctt tattcggcta tgtaggatat caggcactct caaagaggca 7320tataccagta gtcacagaca tatattcaat tgaagatcac aggttggaag acaccacaca 7380cctacagtat gccccgaatg ctatcaagac ggaggggaag gagacagaat tgaaggagct 7440agctcagggg gatgtgcaga gatgtatgga agctatgact aattatgcaa gagatggcat 7500ccaattcatg aagtctcagg cactgaaagt gaaagaaacc cccacttaca aagagacaat 7560ggacaccgtg gcggactatg taaagaagtt catggaggca ctggcggaca gcaaagaaga 7620catcataaaa tatgggttgt gggggacgca cacaacctta tataagagca tcggtgctag 7680gcttgggaac gagactgcgt tcgctaccct ggtcgtgaaa tggctggcat ttgggggaga 7740atcaatagca gaccatgtca aacaagcggc cacagacttg gtcgtttact atatcatcaa 7800cagacctcag ttcccaggag acacggagac acaacaggaa ggaaggaaat ttgtagccag 7860cctactggtc tcagccctgg ctacttacac ttacaaaagc tggaattaca ataatctgtc 7920caagatagtt gaaccggctt tggctactct gccctatgcc gccacagctc tcaagctatt 7980cgcccccact cgattggaga gcgttgtcat actgagtacc gcaatctaca aaacctacct 8040atcaatcagg cgcggaaaaa gcgatggttt gctaggcaca ggggttagtg cggctatgga 8100aatcatgtca caaaacccag tatctgtggg tatagcggtc atgctagggg tgggggccgt 8160agcggcccac aatgcaatcg aagccagtga gcagaagaga acactactca tgaaagtttt 8220tgtaaagaac ttcttggatc aggcagccac tgatgaatta gtcaaggaga gccctgagaa 8280aataataatg gctttgtttg aagcagtgca gacagtcggc aaccctctta gactggtata 8340ccacctttac ggagtttttt acaaagggtg ggaggcaaaa gagttggccc aaaggacagc 8400cggtaggaat cttttcactt tgataatgtt tgaggctgtg gaactactgg gagtagatag 8460cgaaggaaag atccgccagc tatcaagcaa ttacatacta gagctcctgt ataagttccg 8520tgacagtatc aagtccagcg tgaggcagat ggcaatcagc tgggcccctg ccccttttag 8580ttgtgattgg acaccgacgg atgacagaat agggcttccc caagataatt tcctccgagt 8640ggagacaaaa tgcccctgtg gttacaagat gaaagcagtt aagaattgtg ctggggagtt 8700gagactctta gaagaggaag gctcatttct ctgcaggaat aaattcggga gaggttcacg 8760gaactacagg gtgacaaaat actatgatga caatctatca gaaataaagc cagtgataag 8820aatggaagga catgtggaac tctactacaa gggagccact attaaactgg atttcaacaa 8880cagtaaaaca atattggcaa ccgataaatg ggaggtcgat cactccactc tggtcagggt 8940gctcaagagg cacacagggg ctggatatcg tggggcatac ctgggtgaga aaccgaacca 9000caaacatctg atagagaggg actgcgcaac catcaccaaa gataaggttt gttttctcaa 9060gatgaagaga gggtgtgcat ttacttatga cttatccctt cacaacctta cccggctgat 9120cgaattggta cacaagaata acttggaaga caaagagatt cctgccgtta cggtcacaac 9180ctggctggct tacacatttg taaatgaaga tatagggacc ataaaaccag ccttcgggga 9240gaaaataaca ccagagatgc aggaggagat aaccttgcag cctgctgtag tggtggatgc 9300aactgacgtg accgtgaccg tggtagggga aacccctact atgactacag gggagacccc 9360aacaacgttc accagctcag gtccagaccc gaaaggccaa caagttttaa aactgggagt 9420aggtgaaggc caataccccg ggactaatcc acagagagca agcctgcacg aagccataca 9480aagcgcagat gaaaggccct ctgtgttgat attggggtct gataaagcca cctctaatag 9540agtgaaaact gtaaagaatg tgaaggtata cagaggcagg gacccactag aagtgagaga 9600tatgatgagg aggggaaaga tcctagtcat agccctgtct agggttgata atgctctatt 9660gaaatttgta gattacaaag gcacctttct aactagagag accctggagg cattaagttt 9720gggtaggcca aaaaagaaaa acataaccaa ggcagaagca cagtggttgc tgcgcctcga 9780agaccaaatg gaagagctac ccgattggtt cgcagccggg gaacccattt ttttagaggc 9840caatattaaa catgacaggt atcatctggt aggggatata gctactatca aagagaaagc 9900caaacaattg ggggctacag actctacaaa gatatccaag gaggttggtg caaaagtata 9960ttctatgaaa ttgagtaatt gggtgatgca agaagaaaac aaacagagca acttgacccc 10020cttatttgaa gagctcctac agcagtgtcc acccggaggc caaaacaaaa ctgcacatat 10080ggtctctgct taccaactag ctcaagggaa ctggatgcca accagctgcc atgtttttat 10140ggggaccata tctgccagaa ggactaagac ccatccatat gaagcatatg tcaagttaag 10200ggagttggta gaggaacaca agatgaaaac attgtgtccc ggatcaagtc tgcgtaagca 10260caatgaatgg gtaattggca agatcaaata ccagggcaac ctgaggacca aacacatgtt 10320gaaccccggc aaggtggcag agcaactgca cagagaagga cacagacaca atgtgtataa 10380caagacaata ggctcagtga tgacagctac tggcatcagg ttggagaagt tgcccgtggt 10440tagggcccag acagacacaa ccaacttcca ccaagcaata agggataaga tagacaagga 10500agagaatcta cagaccccgg gtttacataa gaaactaatg gaagttttca atgcattgaa 10560acgacccgag ttagagtcct cctatgacgc tgtggaatgg gaggaattgg agagaggaat 10620aaacagaaag ggtgctgctg gtttctttga acgcaaaaac ataggggaga tattggattc 10680agagaaaaat aaagtagaag agattattga caatctgaaa aagggtagaa atatcaaata 10740ctatgaaacc gcaatcccaa aaaatgaaaa gagggatgtc aatgatgact ggaccgcagg 10800tgactttgtg gacgagaaga aacccagagt catacaatac cctgaagcaa aaacaaggct 10860ggccatcacc aaggtgatgt ataagtgggt gaagcagaag ccagtagtca tacccgggta 10920tgaagggaag acacctctgt tccaaatttt tgacaaagta aagaaggaat gggatcaatt 10980ccaaaatcca gtggcagtga gcttcgacac taaggcgtgg gacacccagg tgaccacaaa 11040tgatctggag ctgataaagg acatacaaaa gtactacttc aagaagaaat ggcataaatt 11100tattgacacc ctgactatgc atatgtcaga agtacccgta atcactgctg atggggaggt 11160gtatataagg aaagggcaaa gaggtagtgg acagcccgac acaagcgcag gcaacagcat 11220gctaaatgtg ttaacaatgg tttatgcctt ctgcgaggcc acaggggtac cctacaagag 11280ttttgacagg gtggcaaaaa ttcatgtgtg cggggacgat ggtttcctga tcacagagag 11340agctctcggc gagaaattcg caagcaaggg agtccaaatc ctgtatgaag ctgggaagcc 11400ccagaagatc actgaagggg acaaaatgaa agtggcctac caatttgatg atattgagtt 11460ttgctcccat acaccaatac aagtaaggtg gtcagataac acttctagct acatgccagg 11520gagaaataca accacaatcc tggctaaaat ggccacaagg ttagattcca gtggtgagag 11580gggtaccata gcgtacgaga aagcagtagc attcagcttc ctgctaatgt attcctggaa 11640cccactaatc agaaggattt gcttattggt actatcaact gaactgcaag tgaaaccagg 11700gaagtcaacc acttactatt atgaagggga cccgatatct gcctacaagg aagtcatcgg 11760ccacaatctt ttcgatctca agagaacaag cttcgagaag ctggccaagt taaatctcag 11820catgtccgta ctcggggcct ggactagaca caccagcaaa agactactac aagactgtgt 11880caatatgggt gttaaagagg gcaactggtt agtcaatgca gacagactgg tgagtagtaa 11940gactggaaat aggtatgtac ctggagaagg ccacaccctg caagggagac attatgaaga 12000actggtgttg gcaagaaaac agatcaacag cttccaaggg acagacaggt acaatctagg 12060cccaatagtc aacatggtgt taaggaggct gagagtcatg atgatgaccc tgatagggag 12120aggggtatga gtgcgggtga cccgcgatct ggacccgtca gtaggaccct attgtagata 12180acactaattt tttatttatt tagatattac tatttattta tttatttatt tattgaatga 12240gtaagaactg gtacaaacta cctcatgtta ccacactaca ctcattttaa cagcacttta 12300gctggaagga aaattcctga cgtccacagt tggactaagg taatttccta acggccc 1235723918PRTClassical Swine Fever Virus 2Met Glu Leu Asn His Phe Glu Leu Leu Tyr Lys Thr Asn Lys Gln Lys 1 5 10 15 Pro Met Gly Val Glu Glu Pro Val Tyr Asp Val Thr Gly Arg Pro Leu 20 25 30 Phe Gly Asp Pro Ser Glu Val His Pro Gln Ser Thr Leu Lys Leu Pro 35 40 45 His Asp Arg Gly Arg Gly Asn Ile Lys Thr Thr Leu Lys Asn Leu Pro 50 55 60 Arg Arg Gly Asp Cys Arg Ser Gly Asn His Leu Gly Pro Val Ser Gly 65 70 75 80 Ile Tyr Val Lys Pro Gly Pro Val Phe Tyr Gln Asp Tyr Met Gly Pro 85 90 95 Val Tyr His Arg Ala Pro Leu Glu Phe Phe Asp Glu Ala Gln Phe Cys 100 105 110 Glu Val Thr Lys Arg Ile Gly Arg Val Thr Gly Ser Asp Gly Lys Leu 115 120 125 Tyr His Ile Tyr Val Cys Ile Asp Gly Cys Ile Leu Leu Lys Leu Ala 130 135 140 Lys Arg Gly Glu Pro Arg Thr Leu Lys Trp Ile Arg Asn Leu Thr Asp 145 150 155 160 Cys Pro Leu Trp Val Thr Ser Cys Ser Asp Asp Gly Ala Ser Ala Ser 165 170 175 Lys Glu Lys Lys Pro Asp Arg Ile Asn Lys Gly Lys Leu Lys Ile Ala 180 185 190 Pro Lys Glu His Glu Lys Asp Ser Arg Thr Lys Pro Pro Asp Ala Thr 195 200 205 Ile Val Val Glu Gly Val Lys Tyr Gln Val Lys Lys Lys Gly Lys Val 210 215 220 Lys Gly Lys Asn Thr Gln Asp Gly Leu Tyr His Asn Lys Asn Lys Pro 225 230 235 240 Pro Glu Ser Arg Lys Lys Leu Glu Lys Ala Leu Leu Ala Trp Ala Val 245 250 255 Ile Ala Ile Met Leu Tyr Gln Pro Val Ala Ala Glu Asn Ile Thr Gln 260 265 270 Trp Asn Leu Ser Asp Asn Gly Thr Asn Gly Ile Gln His Ala Met Tyr 275 280 285 Leu Arg Gly Val Ser Arg Ser Leu His Gly Ile Trp Pro Glu Lys Ile 290 295 300 Cys Lys Gly Val Pro Thr Tyr Leu Ala Thr Asp Thr Glu Leu Arg Glu 305 310 315 320 Ile Gln Gly Met Met Asp Ala Ser Glu Gly Thr Asn Tyr Thr Cys Cys 325 330 335 Lys Leu Gln Arg His Glu Trp Asn Lys His Gly Trp Cys Asn Trp Tyr 340 345 350 Asn Ile Asp Pro Trp Ile Gln Leu Met Asn Arg Thr Gln Ala Asn Leu 355 360 365 Ala Glu Gly Pro Pro Ser Lys Glu Cys Ala Val Thr Cys Arg Tyr Asp 370 375 380 Lys Asn Ala Asp Ile Asn Val Val Thr Gln Ala Arg Asn Arg Pro Thr 385 390 395 400 Thr Leu Thr Gly Cys Lys Lys Gly Lys

Asn Phe Ser Phe Ala Gly Thr 405 410 415 Val Ile Glu Gly Pro Cys Asn Phe Asn Val Ser Val Glu Asp Ile Leu 420 425 430 Tyr Gly Asp His Glu Cys Gly Ser Leu Leu Gln Asp Thr Ala Leu Tyr 435 440 445 Leu Val Asp Gly Met Thr Asn Thr Ile Glu Arg Ala Arg Gln Gly Ala 450 455 460 Ala Arg Val Thr Ser Trp Leu Gly Arg Gln Leu Arg Thr Ala Gly Lys 465 470 475 480 Arg Leu Glu Gly Arg Ser Lys Thr Trp Phe Gly Ala Tyr Ala Leu Ser 485 490 495 Pro Tyr Cys Asn Val Thr Ser Lys Ile Gly Tyr Ile Trp Tyr Thr Asn 500 505 510 Asn Cys Thr Pro Ala Cys Leu Pro Lys Asn Thr Lys Ile Ile Gly Pro 515 520 525 Gly Lys Phe Asp Thr Asn Ala Glu Asp Gly Lys Ile Leu His Glu Met 530 535 540 Gly Gly His Leu Ser Glu Phe Leu Leu Leu Ser Leu Val Val Leu Ser 545 550 555 560 Asp Phe Ala Pro Glu Thr Ala Ser Ala Leu Tyr Leu Ile Leu His Tyr 565 570 575 Val Ile Pro Gln Ser His Glu Glu Pro Glu Gly Cys Asp Thr Asn Gln 580 585 590 Leu Asn Leu Thr Val Glu Leu Arg Thr Glu Asp Val Ile Pro Ser Ser 595 600 605 Val Trp Asn Val Gly Lys Tyr Val Cys Val Arg Pro Asp Trp Trp Pro 610 615 620 Tyr Glu Thr Lys Val Ala Leu Leu Phe Glu Glu Ala Gly Gln Val Val 625 630 635 640 Lys Leu Ala Leu Arg Ala Leu Arg Asp Leu Thr Arg Val Trp Asn Ser 645 650 655 Ala Ser Thr Thr Ala Phe Leu Ile Cys Leu Ile Lys Val Leu Arg Gly 660 665 670 Gln Val Val Gln Gly Val Ile Trp Leu Leu Leu Val Thr Gly Ala Pro 675 680 685 Val Ser Cys Thr His Leu Ala Ala Ala Asp Tyr Lys Asp Asp Asp Asp 690 695 700 Lys Gly Ala Gln Gly Arg Leu Ala Cys Lys Glu Asp His Arg Tyr Ala 705 710 715 720 Ile Ser Thr Thr Asn Glu Ile Gly Leu Leu Gly Ala Glu Gly Leu Thr 725 730 735 Thr Thr Trp Lys Glu Tyr Asn His Asn Leu Gln Leu Asp Asp Gly Thr 740 745 750 Val Lys Ala Ile Cys Met Ala Gly Ser Phe Lys Val Thr Ala Leu Asn 755 760 765 Val Val Ser Arg Arg Tyr Leu Ala Ser Leu His Lys Asp Ala Leu Pro 770 775 780 Thr Ser Val Thr Phe Glu Leu Leu Phe Asp Gly Thr Ser Pro Leu Thr 785 790 795 800 Glu Glu Met Gly Asp Asp Phe Gly Phe Gly Leu Cys Pro Tyr Asp Thr 805 810 815 Ser Pro Val Val Lys Gly Lys Tyr Asn Thr Thr Leu Leu Asn Gly Ser 820 825 830 Ala Phe Tyr Leu Val Cys Pro Ile Gly Trp Thr Gly Val Ile Glu Cys 835 840 845 Thr Ser Phe Gly Met Asp Thr Leu Arg Thr Glu Val Val Lys Thr Phe 850 855 860 Arg Arg Glu Lys Pro Phe Pro Tyr Arg Arg Asp Cys Val Thr Thr Thr 865 870 875 880 Val Glu Asn Glu Asp Leu Phe Tyr Cys Lys Trp Gly Gly Asn Trp Thr 885 890 895 Cys Val Lys Gly Glu Pro Val Thr Tyr Thr Gly Gly Pro Val Lys Gln 900 905 910 Cys Arg Trp Cys Gly Phe Asp Phe Asn Glu Pro Asp Gly Leu Pro His 915 920 925 Tyr Pro Ile Gly Lys Cys Ile Leu Ala Asn Glu Thr Gly Tyr Arg Ile 930 935 940 Val Asp Ser Thr Asp Cys Asn Arg Asp Gly Val Val Ile Ser Thr Glu 945 950 955 960 Gly Ser His Glu Cys Leu Ile Gly Asn Thr Thr Val Lys Val His Ala 965 970 975 Leu Asp Glu Arg Leu Gly Pro Met Pro Cys Arg Pro Lys Glu Ile Val 980 985 990 Ser Ser Ala Gly Pro Val Arg Lys Thr Ser Cys Thr Phe Asn Tyr Ala 995 1000 1005 Lys Thr Leu Arg Asn Arg Tyr Tyr Glu Pro Arg Asp Ser Tyr Phe 1010 1015 1020 Gln Gln Tyr Met Leu Lys Gly Glu Tyr Gln Tyr Trp Phe Asp Leu 1025 1030 1035 Asp Val Thr Asp Arg His Ser Asp Tyr Phe Ala Glu Phe Ile Val 1040 1045 1050 Leu Val Val Val Ala Leu Leu Gly Gly Arg Tyr Val Leu Trp Leu 1055 1060 1065 Ile Val Thr Tyr Ile Val Leu Thr Glu Gln Leu Ala Ala Gly Leu 1070 1075 1080 Gln Leu Gly Gln Gly Glu Val Val Leu Ile Gly Asn Leu Ile Thr 1085 1090 1095 His Thr Asp Ile Glu Val Val Val Tyr Phe Leu Leu Leu Tyr Leu 1100 1105 1110 Val Met Arg Asp Glu Pro Ile Lys Lys Trp Ile Leu Leu Leu Phe 1115 1120 1125 His Ala Met Thr Asn Asn Pro Val Lys Thr Ile Thr Val Ala Leu 1130 1135 1140 Leu Met Val Ser Gly Val Ala Lys Gly Gly Lys Ile Asp Gly Gly 1145 1150 1155 Trp Gln Arg Leu Pro Glu Thr Asn Phe Asp Ile Gln Leu Ala Leu 1160 1165 1170 Thr Val Ile Val Val Ala Val Met Leu Leu Ala Lys Lys Asp Pro 1175 1180 1185 Thr Thr Val Pro Leu Val Ile Thr Val Ala Thr Leu Arg Thr Ala 1190 1195 1200 Lys Ile Thr Asn Gly Leu Ser Thr Asp Leu Ala Ile Ala Thr Val 1205 1210 1215 Ser Thr Ala Leu Leu Thr Trp Thr Tyr Ile Ser Asp Tyr Tyr Lys 1220 1225 1230 Tyr Lys Thr Leu Leu Gln Tyr Leu Ile Ser Thr Val Thr Gly Ile 1235 1240 1245 Phe Leu Ile Arg Val Leu Lys Gly Val Gly Glu Leu Asp Leu His 1250 1255 1260 Thr Pro Thr Leu Pro Ser Tyr Arg Pro Leu Phe Phe Ile Leu Val 1265 1270 1275 Tyr Leu Ile Ser Thr Ala Val Val Thr Arg Trp Asn Leu Asp Ile 1280 1285 1290 Ala Gly Leu Leu Leu Gln Cys Val Pro Thr Leu Leu Met Val Phe 1295 1300 1305 Thr Met Trp Ala Asp Ile Leu Thr Leu Ile Leu Ile Leu Pro Thr 1310 1315 1320 Tyr Glu Leu Thr Lys Leu Tyr Tyr Leu Lys Glu Val Lys Ile Gly 1325 1330 1335 Ala Glu Arg Gly Trp Leu Trp Lys Thr Asn Phe Lys Arg Val Asn 1340 1345 1350 Asp Ile Tyr Glu Val Asp Gln Ala Gly Glu Gly Val Tyr Leu Phe 1355 1360 1365 Pro Ser Lys Gln Lys Thr Gly Thr Ile Thr Gly Thr Met Leu Pro 1370 1375 1380 Leu Ile Lys Ala Ile Leu Ile Ser Cys Ile Ser Asn Lys Trp Gln 1385 1390 1395 Phe Ile Tyr Leu Leu Tyr Leu Ile Phe Glu Val Ser Tyr Tyr Leu 1400 1405 1410 His Lys Lys Ile Ile Asp Glu Ile Ala Gly Gly Thr Asn Phe Ile 1415 1420 1425 Ser Arg Leu Val Ala Ala Leu Ile Glu Ala Asn Trp Ala Phe Asp 1430 1435 1440 Asn Glu Glu Val Arg Gly Leu Lys Lys Phe Phe Leu Leu Ser Ser 1445 1450 1455 Arg Val Lys Glu Leu Ile Ile Lys His Lys Val Arg Asn Glu Val 1460 1465 1470 Met Val His Trp Phe Gly Asp Glu Glu Val Tyr Gly Met Pro Lys 1475 1480 1485 Leu Val Gly Leu Val Lys Ala Ala Thr Leu Ser Lys Asn Lys His 1490 1495 1500 Cys Ile Leu Cys Thr Val Cys Glu Asn Arg Glu Trp Arg Gly Glu 1505 1510 1515 Thr Cys Pro Lys Cys Gly Arg Phe Gly Pro Pro Val Thr Cys Gly 1520 1525 1530 Met Thr Leu Ala Asp Phe Glu Glu Lys His Tyr Lys Arg Ile Phe 1535 1540 1545 Phe Arg Glu Asp Gln Ser Glu Gly Pro Val Arg Glu Glu Tyr Ala 1550 1555 1560 Gly Tyr Leu Gln Tyr Arg Ala Arg Gly Gln Leu Phe Leu Arg Asn 1565 1570 1575 Leu Pro Val Leu Ala Thr Lys Val Lys Met Leu Leu Val Gly Asn 1580 1585 1590 Leu Gly Thr Glu Val Gly Asp Leu Glu His Leu Gly Trp Val Leu 1595 1600 1605 Arg Gly Pro Ala Val Cys Lys Lys Val Thr Glu His Glu Lys Cys 1610 1615 1620 Thr Thr Ser Ile Met Asp Lys Leu Thr Ala Phe Phe Gly Val Met 1625 1630 1635 Pro Arg Gly Thr Thr Pro Arg Ala Pro Val Arg Phe Pro Thr Ser 1640 1645 1650 Leu Leu Lys Ile Arg Arg Gly Leu Glu Thr Gly Trp Ala Tyr Thr 1655 1660 1665 His Gln Gly Gly Ile Ser Ser Val Asp His Val Thr Cys Gly Lys 1670 1675 1680 Asp Leu Leu Val Cys Asp Thr Met Gly Arg Thr Arg Val Val Cys 1685 1690 1695 Gln Ser Asn Asn Lys Met Thr Asp Glu Ser Glu Tyr Gly Val Lys 1700 1705 1710 Thr Asp Ser Gly Cys Pro Glu Gly Ala Arg Cys Tyr Val Phe Asn 1715 1720 1725 Pro Glu Ala Val Asn Ile Ser Gly Thr Lys Gly Ala Met Val His 1730 1735 1740 Leu Gln Lys Thr Gly Gly Glu Phe Thr Cys Val Thr Ala Ser Gly 1745 1750 1755 Thr Pro Ala Phe Phe Asp Leu Lys Asn Leu Lys Gly Trp Ser Gly 1760 1765 1770 Leu Pro Ile Phe Glu Ala Ser Ser Gly Arg Val Val Gly Arg Val 1775 1780 1785 Lys Val Gly Lys Asn Glu Asp Ser Lys Pro Thr Lys Leu Met Ser 1790 1795 1800 Gly Ile Gln Thr Val Ser Lys Ser Thr Thr Asp Leu Thr Glu Met 1805 1810 1815 Val Lys Lys Ile Thr Thr Met Asn Arg Gly Glu Phe Arg Gln Ile 1820 1825 1830 Thr Leu Ala Thr Gly Ala Gly Lys Thr Thr Glu Leu Pro Arg Ser 1835 1840 1845 Val Ile Glu Glu Ile Gly Arg His Lys Arg Val Leu Val Leu Ile 1850 1855 1860 Pro Leu Arg Ala Ala Ala Glu Ser Val Tyr Gln Tyr Met Arg Gln 1865 1870 1875 Lys His Pro Ser Ile Ala Phe Asn Leu Arg Ile Gly Glu Met Lys 1880 1885 1890 Glu Gly Asp Met Ala Thr Gly Ile Thr Tyr Ala Ser Tyr Gly Tyr 1895 1900 1905 Phe Cys Gln Met Pro Gln Pro Lys Leu Arg Ala Ala Met Val Glu 1910 1915 1920 Tyr Ser Phe Ile Phe Leu Asp Glu Tyr His Cys Ala Thr Pro Glu 1925 1930 1935 Gln Leu Ala Ile Met Gly Lys Ile His Arg Phe Ser Glu Asn Leu 1940 1945 1950 Arg Val Val Ala Met Thr Ala Thr Pro Ala Gly Thr Val Thr Thr 1955 1960 1965 Thr Gly Gln Lys His Pro Ile Glu Glu Tyr Ile Ala Pro Glu Val 1970 1975 1980 Met Lys Gly Glu Asp Leu Gly Ser Glu Tyr Leu Asp Ile Ala Gly 1985 1990 1995 Leu Lys Ile Pro Val Glu Glu Met Lys Ser Asn Met Leu Val Phe 2000 2005 2010 Val Pro Thr Arg Asn Met Ala Val Glu Thr Ala Lys Lys Leu Lys 2015 2020 2025 Ala Lys Gly Tyr Asn Ser Gly Tyr Tyr Tyr Ser Gly Glu Asp Pro 2030 2035 2040 Ser Asn Leu Arg Val Val Thr Ser Gln Ser Pro Tyr Val Val Val 2045 2050 2055 Ala Thr Asn Ala Ile Glu Ser Gly Val Thr Leu Pro Asp Leu Asp 2060 2065 2070 Val Val Val Asp Thr Gly Leu Lys Cys Glu Lys Arg Ile Arg Leu 2075 2080 2085 Ser Pro Lys Met Pro Phe Ile Val Thr Gly Leu Lys Arg Met Ala 2090 2095 2100 Val Thr Ile Gly Glu Gln Ala Gln Arg Arg Gly Arg Val Gly Arg 2105 2110 2115 Val Lys Pro Gly Arg Tyr Tyr Arg Ser Gln Glu Thr Pro Val Gly 2120 2125 2130 Ser Lys Asp Tyr His Tyr Asp Leu Leu Gln Ala Gln Arg Tyr Gly 2135 2140 2145 Ile Glu Asp Gly Ile Asn Ile Thr Lys Ser Phe Arg Glu Met Asn 2150 2155 2160 Tyr Asp Trp Ser Leu Tyr Glu Glu Asp Ser Leu Met Ile Thr Gln 2165 2170 2175 Leu Glu Ile Leu Asn Asn Leu Leu Ile Ser Glu Glu Leu Pro Met 2180 2185 2190 Ala Val Lys Asn Ile Met Ala Arg Thr Asp His Pro Glu Pro Ile 2195 2200 2205 Gln Leu Ala Tyr Asn Ser Tyr Glu Thr Gln Val Pro Val Leu Phe 2210 2215 2220 Pro Lys Ile Lys Asn Gly Glu Val Thr Asp Ser Tyr Asp Asn Tyr 2225 2230 2235 Thr Phe Leu Asn Ala Arg Lys Leu Gly Asp Asp Val Pro Pro Tyr 2240 2245 2250 Val Tyr Ala Thr Glu Asp Glu Asp Leu Ala Val Glu Leu Leu Gly 2255 2260 2265 Leu Asp Trp Pro Asp Pro Gly Asn Gln Gly Thr Val Glu Ala Gly 2270 2275 2280 Arg Ala Leu Lys Gln Val Val Gly Leu Ser Thr Ala Glu Asn Ala 2285 2290 2295 Leu Leu Val Ala Leu Phe Gly Tyr Val Gly Tyr Gln Ala Leu Ser 2300 2305 2310 Lys Arg His Ile Pro Val Val Thr Asp Ile Tyr Ser Ile Glu Asp 2315 2320 2325 His Arg Leu Glu Asp Thr Thr His Leu Gln Tyr Ala Pro Asn Ala 2330 2335 2340 Ile Lys Thr Glu Gly Lys Glu Thr Glu Leu Lys Glu Leu Ala Gln 2345 2350 2355 Gly Asp Val Gln Arg Cys Met Glu Ala Met Thr Asn Tyr Ala Arg 2360 2365 2370 Asp Gly Ile Gln Phe Met Lys Ser Gln Ala Leu Lys Val Lys Glu 2375 2380 2385 Thr Pro Thr Tyr Lys Glu Thr Met Asp Thr Val Ala Asp Tyr Val 2390 2395 2400 Lys Lys Phe Met Glu Ala Leu Ala Asp Ser Lys Glu Asp Ile Ile 2405 2410 2415 Lys Tyr Gly Leu Trp Gly Thr His Thr Thr Leu Tyr Lys Ser Ile 2420 2425 2430 Gly Ala Arg Leu Gly Asn Glu Thr Ala Phe Ala Thr Leu Val Val 2435 2440 2445 Lys Trp Leu Ala Phe Gly Gly Glu Ser Ile Ala Asp His Val Lys 2450 2455 2460 Gln Ala Ala Thr Asp Leu Val Val Tyr Tyr Ile Ile Asn Arg Pro 2465 2470 2475 Gln Phe Pro Gly Asp Thr Glu Thr Gln Gln Glu Gly Arg Lys Phe 2480 2485 2490 Val Ala Ser Leu Leu Val Ser Ala Leu Ala Thr Tyr Thr Tyr Lys 2495 2500 2505 Ser Trp Asn Tyr Asn Asn Leu Ser Lys Ile Val Glu Pro Ala Leu 2510 2515 2520 Ala Thr Leu Pro Tyr Ala Ala Thr Ala Leu Lys Leu Phe Ala Pro 2525 2530 2535 Thr Arg Leu Glu Ser Val Val Ile Leu Ser Thr Ala Ile Tyr Lys 2540 2545 2550 Thr Tyr Leu Ser Ile Arg Arg Gly Lys Ser Asp Gly Leu Leu Gly 2555 2560 2565 Thr Gly Val Ser Ala Ala Met Glu Ile Met Ser Gln Asn Pro Val 2570 2575 2580 Ser Val Gly Ile Ala Val Met Leu Gly Val Gly Ala Val Ala Ala 2585 2590 2595 His Asn Ala Ile Glu Ala Ser Glu Gln Lys Arg Thr Leu Leu Met 2600 2605 2610 Lys Val Phe Val Lys Asn Phe Leu Asp Gln Ala Ala Thr Asp Glu 2615 2620 2625

Leu Val Lys Glu Ser Pro Glu Lys Ile Ile Met Ala Leu Phe Glu 2630 2635 2640 Ala Val Gln Thr Val Gly Asn Pro Leu Arg Leu Val Tyr His Leu 2645 2650 2655 Tyr Gly Val Phe Tyr Lys Gly Trp Glu Ala Lys Glu Leu Ala Gln 2660 2665 2670 Arg Thr Ala Gly Arg Asn Leu Phe Thr Leu Ile Met Phe Glu Ala 2675 2680 2685 Val Glu Leu Leu Gly Val Asp Ser Glu Gly Lys Ile Arg Gln Leu 2690 2695 2700 Ser Ser Asn Tyr Ile Leu Glu Leu Leu Tyr Lys Phe Arg Asp Ser 2705 2710 2715 Ile Lys Ser Ser Val Arg Gln Met Ala Ile Ser Trp Ala Pro Ala 2720 2725 2730 Pro Phe Ser Cys Asp Trp Thr Pro Thr Asp Asp Arg Ile Gly Leu 2735 2740 2745 Pro Gln Asp Asn Phe Leu Arg Val Glu Thr Lys Cys Pro Cys Gly 2750 2755 2760 Tyr Lys Met Lys Ala Val Lys Asn Cys Ala Gly Glu Leu Arg Leu 2765 2770 2775 Leu Glu Glu Glu Gly Ser Phe Leu Cys Arg Asn Lys Phe Gly Arg 2780 2785 2790 Gly Ser Arg Asn Tyr Arg Val Thr Lys Tyr Tyr Asp Asp Asn Leu 2795 2800 2805 Ser Glu Ile Lys Pro Val Ile Arg Met Glu Gly His Val Glu Leu 2810 2815 2820 Tyr Tyr Lys Gly Ala Thr Ile Lys Leu Asp Phe Asn Asn Ser Lys 2825 2830 2835 Thr Ile Leu Ala Thr Asp Lys Trp Glu Val Asp His Ser Thr Leu 2840 2845 2850 Val Arg Val Leu Lys Arg His Thr Gly Ala Gly Tyr Arg Gly Ala 2855 2860 2865 Tyr Leu Gly Glu Lys Pro Asn His Lys His Leu Ile Glu Arg Asp 2870 2875 2880 Cys Ala Thr Ile Thr Lys Asp Lys Val Cys Phe Leu Lys Met Lys 2885 2890 2895 Arg Gly Cys Ala Phe Thr Tyr Asp Leu Ser Leu His Asn Leu Thr 2900 2905 2910 Arg Leu Ile Glu Leu Val His Lys Asn Asn Leu Glu Asp Lys Glu 2915 2920 2925 Ile Pro Ala Val Thr Val Thr Thr Trp Leu Ala Tyr Thr Phe Val 2930 2935 2940 Asn Glu Asp Ile Gly Thr Ile Lys Pro Ala Phe Gly Glu Lys Ile 2945 2950 2955 Thr Pro Glu Met Gln Glu Glu Ile Thr Leu Gln Pro Ala Val Val 2960 2965 2970 Val Asp Ala Thr Asp Val Thr Val Thr Val Val Gly Glu Thr Pro 2975 2980 2985 Thr Met Thr Thr Gly Glu Thr Pro Thr Thr Phe Thr Ser Ser Gly 2990 2995 3000 Pro Asp Pro Lys Gly Gln Gln Val Leu Lys Leu Gly Val Gly Glu 3005 3010 3015 Gly Gln Tyr Pro Gly Thr Asn Pro Gln Arg Ala Ser Leu His Glu 3020 3025 3030 Ala Ile Gln Ser Ala Asp Glu Arg Pro Ser Val Leu Ile Leu Gly 3035 3040 3045 Ser Asp Lys Ala Thr Ser Asn Arg Val Lys Thr Val Lys Asn Val 3050 3055 3060 Lys Val Tyr Arg Gly Arg Asp Pro Leu Glu Val Arg Asp Met Met 3065 3070 3075 Arg Arg Gly Lys Ile Leu Val Ile Ala Leu Ser Arg Val Asp Asn 3080 3085 3090 Ala Leu Leu Lys Phe Val Asp Tyr Lys Gly Thr Phe Leu Thr Arg 3095 3100 3105 Glu Thr Leu Glu Ala Leu Ser Leu Gly Arg Pro Lys Lys Lys Asn 3110 3115 3120 Ile Thr Lys Ala Glu Ala Gln Trp Leu Leu Arg Leu Glu Asp Gln 3125 3130 3135 Met Glu Glu Leu Pro Asp Trp Phe Ala Ala Gly Glu Pro Ile Phe 3140 3145 3150 Leu Glu Ala Asn Ile Lys His Asp Arg Tyr His Leu Val Gly Asp 3155 3160 3165 Ile Ala Thr Ile Lys Glu Lys Ala Lys Gln Leu Gly Ala Thr Asp 3170 3175 3180 Ser Thr Lys Ile Ser Lys Glu Val Gly Ala Lys Val Tyr Ser Met 3185 3190 3195 Lys Leu Ser Asn Trp Val Met Gln Glu Glu Asn Lys Gln Ser Asn 3200 3205 3210 Leu Thr Pro Leu Phe Glu Glu Leu Leu Gln Gln Cys Pro Pro Gly 3215 3220 3225 Gly Gln Asn Lys Thr Ala His Met Val Ser Ala Tyr Gln Leu Ala 3230 3235 3240 Gln Gly Asn Trp Met Pro Thr Ser Cys His Val Phe Met Gly Thr 3245 3250 3255 Ile Ser Ala Arg Arg Thr Lys Thr His Pro Tyr Glu Ala Tyr Val 3260 3265 3270 Lys Leu Arg Glu Leu Val Glu Glu His Lys Met Lys Thr Leu Cys 3275 3280 3285 Pro Gly Ser Ser Leu Arg Lys His Asn Glu Trp Val Ile Gly Lys 3290 3295 3300 Ile Lys Tyr Gln Gly Asn Leu Arg Thr Lys His Met Leu Asn Pro 3305 3310 3315 Gly Lys Val Ala Glu Gln Leu His Arg Glu Gly His Arg His Asn 3320 3325 3330 Val Tyr Asn Lys Thr Ile Gly Ser Val Met Thr Ala Thr Gly Ile 3335 3340 3345 Arg Leu Glu Lys Leu Pro Val Val Arg Ala Gln Thr Asp Thr Thr 3350 3355 3360 Asn Phe His Gln Ala Ile Arg Asp Lys Ile Asp Lys Glu Glu Asn 3365 3370 3375 Leu Gln Thr Pro Gly Leu His Lys Lys Leu Met Glu Val Phe Asn 3380 3385 3390 Ala Leu Lys Arg Pro Glu Leu Glu Ser Ser Tyr Asp Ala Val Glu 3395 3400 3405 Trp Glu Glu Leu Glu Arg Gly Ile Asn Arg Lys Gly Ala Ala Gly 3410 3415 3420 Phe Phe Glu Arg Lys Asn Ile Gly Glu Ile Leu Asp Ser Glu Lys 3425 3430 3435 Asn Lys Val Glu Glu Ile Ile Asp Asn Leu Lys Lys Gly Arg Asn 3440 3445 3450 Ile Lys Tyr Tyr Glu Thr Ala Ile Pro Lys Asn Glu Lys Arg Asp 3455 3460 3465 Val Asn Asp Asp Trp Thr Ala Gly Asp Phe Val Asp Glu Lys Lys 3470 3475 3480 Pro Arg Val Ile Gln Tyr Pro Glu Ala Lys Thr Arg Leu Ala Ile 3485 3490 3495 Thr Lys Val Met Tyr Lys Trp Val Lys Gln Lys Pro Val Val Ile 3500 3505 3510 Pro Gly Tyr Glu Gly Lys Thr Pro Leu Phe Gln Ile Phe Asp Lys 3515 3520 3525 Val Lys Lys Glu Trp Asp Gln Phe Gln Asn Pro Val Ala Val Ser 3530 3535 3540 Phe Asp Thr Lys Ala Trp Asp Thr Gln Val Thr Thr Asn Asp Leu 3545 3550 3555 Glu Leu Ile Lys Asp Ile Gln Lys Tyr Tyr Phe Lys Lys Lys Trp 3560 3565 3570 His Lys Phe Ile Asp Thr Leu Thr Met His Met Ser Glu Val Pro 3575 3580 3585 Val Ile Thr Ala Asp Gly Glu Val Tyr Ile Arg Lys Gly Gln Arg 3590 3595 3600 Gly Ser Gly Gln Pro Asp Thr Ser Ala Gly Asn Ser Met Leu Asn 3605 3610 3615 Val Leu Thr Met Val Tyr Ala Phe Cys Glu Ala Thr Gly Val Pro 3620 3625 3630 Tyr Lys Ser Phe Asp Arg Val Ala Lys Ile His Val Cys Gly Asp 3635 3640 3645 Asp Gly Phe Leu Ile Thr Glu Arg Ala Leu Gly Glu Lys Phe Ala 3650 3655 3660 Ser Lys Gly Val Gln Ile Leu Tyr Glu Ala Gly Lys Pro Gln Lys 3665 3670 3675 Ile Thr Glu Gly Asp Lys Met Lys Val Ala Tyr Gln Phe Asp Asp 3680 3685 3690 Ile Glu Phe Cys Ser His Thr Pro Ile Gln Val Arg Trp Ser Asp 3695 3700 3705 Asn Thr Ser Ser Tyr Met Pro Gly Arg Asn Thr Thr Thr Ile Leu 3710 3715 3720 Ala Lys Met Ala Thr Arg Leu Asp Ser Ser Gly Glu Arg Gly Thr 3725 3730 3735 Ile Ala Tyr Glu Lys Ala Val Ala Phe Ser Phe Leu Leu Met Tyr 3740 3745 3750 Ser Trp Asn Pro Leu Ile Arg Arg Ile Cys Leu Leu Val Leu Ser 3755 3760 3765 Thr Glu Leu Gln Val Lys Pro Gly Lys Ser Thr Thr Tyr Tyr Tyr 3770 3775 3780 Glu Gly Asp Pro Ile Ser Ala Tyr Lys Glu Val Ile Gly His Asn 3785 3790 3795 Leu Phe Asp Leu Lys Arg Thr Ser Phe Glu Lys Leu Ala Lys Leu 3800 3805 3810 Asn Leu Ser Met Ser Val Leu Gly Ala Trp Thr Arg His Thr Ser 3815 3820 3825 Lys Arg Leu Leu Gln Asp Cys Val Asn Met Gly Val Lys Glu Gly 3830 3835 3840 Asn Trp Leu Val Asn Ala Asp Arg Leu Val Ser Ser Lys Thr Gly 3845 3850 3855 Asn Arg Tyr Val Pro Gly Glu Gly His Thr Leu Gln Gly Arg His 3860 3865 3870 Tyr Glu Glu Leu Val Leu Ala Arg Lys Gln Ile Asn Ser Phe Gln 3875 3880 3885 Gly Thr Asp Arg Tyr Asn Leu Gly Pro Ile Val Asn Met Val Leu 3890 3895 3900 Arg Arg Leu Arg Val Met Met Met Thr Leu Ile Gly Arg Gly Val 3905 3910 3915 312357DNAClassical Swine Fever Virus 3gtatacgagg ttagttcatt ctcgtgtaca tgattggaca aatcaaaatc tcaatttggt 60tcagggcctc cctccagcga cggccgagct gggctagcca tgcccacagt aggactagca 120aacggaggga ctagccgtag tggcgagctc cctgggtggt ctaagtcctg agtacaggac 180agtcgtcagt agttcgacgt gagcagaagc ccacctcgag atgctatgtg gacgagggca 240tgcccaagac acaccttaac cctagcgggg gtcgttaggg tgaaatcaca ccatgtgatg 300ggagtacgac ctgatagggt gctgcagagg cccactatta ggctagtata aaaatctctg 360ctgtacatgg cacatggagt tgaatcattt tgaactttta tacaaaacaa acaaacaaaa 420accaatggga gtggaggaac cggtatacga tgtaacgggg agaccattgt ttggagaccc 480aagtgaggta cacccacaat caacattgaa gctaccacat gataggggga gaggcaacat 540caaaacaaca ctgaagaacc tacctaggag aggtgactgc aggagtggca accacctagg 600cccggttagt gggatatatg taaagcccgg ccctgtcttt tatcaggact acatgggccc 660agtctatcat agagcccctc tagagttttt tgacgaagca cagttttgtg aggtgaccaa 720aaggataggt agggtgacag gtagtgacgg aaagctttac catatatacg tgtgcatcga 780tggttgcatc ctgctgaagc tagccaagag gggcgagcca agaaccctga agtggattag 840aaatctcacc gactgtccat tgtgggttac cagttgttct gatgatggtg caagtgcaag 900taaagagaag aaaccagata ggatcaacaa gggtaaatta aagatagccc caaaagagca 960tgagaaggac agcaggacta agccacctga tgctacgatt gtagtggaag gagtaaaata 1020ccaggtcaaa aagaaaggta aagttaaggg aaagaatacc caagacggcc tgtaccacaa 1080caagaataaa ccaccagaat ctaggaagaa attagaaaaa gccctattgg catgggcagt 1140gatagcaatt atgttatacc aacctgttgc agccgaaaat ataactcaat ggaacctgag 1200tgacaacggt accaatggta tccagcacgc tatgtacctt agaggagtca gcagaagctt 1260gcatgggatc tggccagaaa aaatatgcaa aggagtcccc acctacctgg ccacagacac 1320ggaactgaga gaaatacagg gaatgatgga tgccagcgag gggacaaact atacgtgctg 1380taagttacag agacatgaat ggaacaaaca tggatggtgt aactggtata acatagaccc 1440ctggatacag ttgatgaata gaacccaagc aaacttggca gaaggccctc cgagcaagga 1500gtgcgccgtg acttgcaggt acgataaaaa tgctgacatt aacgtggtca cccaggccag 1560aaacaggcca accaccctaa ctggctgcaa gaaagggaaa aatttttctt ttgcgggtac 1620agttatagag ggcccatgta atttcaacgt ttctgttgag gatatcttat atggggatca 1680tgagtgtggc agtctactcc aggatacggc tctataccta gtagatggaa tgaccaacac 1740tatagagaga gccaggcagg gagccgcgag ggtgacatct tggctaggga ggcaactcag 1800aactgccggg aagaggttgg agggcagaag caaaacctgg tttggtgcct atgccctatc 1860accttattgt aatgtgacaa gcaaaatagg gtacatatgg tacactaaca actgtacccc 1920ggcttgcctc cccaaaaata caaagataat aggccccggt aaatttgaca ctaacgcgga 1980agacggaaag attctccatg agatgggggg ccacctatca gaatttctgc tgctctctct 2040ggtcgttctg tctgacttcg cccctgaaac agccagcgcg ttatacctca ttttgcacta 2100cgtgatccct caatcccatg aagaacctga aggctgtgac acaaaccagc tgaatttaac 2160agtggaactc aggactgaag acgtgatacc atcatcagtc tggaatgttg gcaaatatgt 2220gtgtgttaga ccagactggt ggccatatga aaccaaggtg gctttgttat ttgaagaggc 2280aggacaggtc gtaaagttag ccttgcgggc actgagggat ttaaccaggg tctggaatag 2340cgcatcaacc acggcattcc tcatctgctt gataaaagta ttaagaggac aggtcgtgca 2400aggtgtgata tggctgttac tggtaactgg ggcacctgtc tcttgtacac atcttgcggc 2460cgcagattac aaggatgacg acgataaggg ggcacaaggc cggctagcct gcaaggaaga 2520tcacaggtac gctatatcaa caaccaatga gatagggcta cttggggccg aaggtctcac 2580taccacctgg aaagaataca accacaattt gcaactggat gatgggaccg tcaaggccat 2640ctgcatggca ggttccttta aagtcacagc acttaatgtg gttagtagga ggtatctggc 2700atcattacat aaggacgctt tacccacttc cgtgacattc gagctcctgt tcgacgggac 2760cagcccattg accgaggaaa tgggagatga cttcgggttc ggactgtgtc cgtatgatac 2820gagccctgta gtcaagggaa agtacaacac aaccttgttg aatggtagtg cattctacct 2880agtttgccca atagggtgga cgggtgttat agagtgcacg tcatttaata tggacactct 2940gagaacagaa gtggtaaaga ccttcagaag agagaaaccc tttccgtaca gaagggattg 3000tgtgaccact acagtggaaa atgaagatct attctactgt aaatgggggg gcaattggac 3060atgtgtgaaa ggtgaaccag tgacctacac gggggggcca gtaaaacaat gcagatggtg 3120tggcttcgac ttcaatgagc ctgacggact cccacactac cccataggta agtgcatttt 3180ggcaaatgag acaggttaca gaatagtgga ttcaacggac tgtaacagag atggcgttgt 3240aatcagcaca gaggggagtc atgagtgctt gattggtaac acaactgtca aggtgcatgc 3300attagatgaa agactaggcc ctatgccatg caggcctaag gagatcgtct ctagtgcggg 3360acctgtaagg aaaacttcct gtacattcaa ctacgcaaaa actctgagga acaggtatta 3420tgagcccagg gacagctatt tccaacaata tatgctcaag ggcgagtatc agtactggtt 3480tgatctggat gtgaccgacc gccactcaga ttacttcgca gaattcattg tcttggtggt 3540ggtggcactg ttgggaggaa gatatgtcct gtggctaata gtgacctaca tagttctaac 3600agaacaactc gccgctggtc tacagttagg ccagggtgag gtagtgttaa tagggaactt 3660aatcacccac acagatattg aggttgtagt atatttctta ctgctctatt tggtcatgag 3720agatgagcct ataaagaaat ggatactact gctgttccat gctatgacca acaatccagt 3780taagaccata acagtggcac tgctcatggt tagcggggtt gccaagggtg gaaagataga 3840tggtggttgg cagcggctgc cggagaccaa ctttgatatc caactcgcgc tgacagttat 3900agtagtcgct gtgatgttgc tggcaaagaa agatccgact accgtcccct tggttataac 3960ggtggcaacc ctgagaacgg ctaagataac taatggactt agtacagatc tagccatagc 4020tacagtgtca acagctttgc taacctggac ctacattagt gactattata aatacaagac 4080cttgctacag taccttatta gcacagtgac aggtatcttc ttgataaggg tactgaaggg 4140ggtaggtgag ttagatttac acaccccaac cttaccatct tacagacccc tcttcttcat 4200cctcgtgtac ctcatttcca ctgcagtggt aacaagatgg aatctggaca tagccggatt 4260gctgctgcag tgtgtcccaa cccttttaat ggttttcacg atgtgggcag acatccttac 4320cctgatcctc atactgccta cttacgagtt gacaaaacta tattacctca aggaagtgaa 4380gattggggca gaaaggggct ggttgtggaa gaccaacttc aagagggtaa atgacatata 4440cgaagttgac caagctggtg agggggtgta ccttttccca tcaaaacaaa agacaggtac 4500aataacaggt actatgttgc cattgatcaa agccatactc ataagttgca tcagcaataa 4560gtggcaattt atatatctat tgtacttgat attcgaagtg tcttactacc ttcacaagaa 4620gatcatagat gaaatagcag gagggaccaa cttcatctcg agacttgtag ccgctctgat 4680tgaagccaat tgggcctttg acaacgaaga agttagaggt ttaaagaagt tcttcctgct 4740gtctagtagg gttaaagaac tgatcatcaa acacaaagtg aggaatgaag tgatggtcca 4800ctggtttggc gacgaagagg tctatgggat gccgaagctg gttggcttag tcaaggcagc 4860aacactgagt aaaaataaac attgtatttt gtgcaccgtc tgtgaaaaca gagagtggag 4920aggagaaacc tgcccaaaat gcggccgttt tgggccacca gtgacctgtg gcatgaccct 4980agccgacttt gaagaaaaac actataagag gattttcttt agagaggatc aatcagaagg 5040gccggttagg gaggagtatg cagggtatct gcaatataga gccagagggc aattattcct 5100gaggaatctc ccggtgctag caacaaaagt caagatgctc ctggtcggaa atcttgggac 5160ggaggtgggg gatttggaac accttggctg ggtgctcaga gggcctgccg tttgcaagaa 5220ggttaccgaa catgagaaat gcaccacatc cataatggac aaattaactg ctttcttcgg 5280tgttatgcca aggggcacca cacctagagc ccctgtgaga ttccccacct ctctcttaaa 5340gataagaagg gggctggaaa ctggctgggc gtacacacac caaggtggca tcagttcagt 5400ggaccatgtc acttgtggga aagacttact ggtatgtgac actatgggcc ggacaagggt 5460tgtttgccaa tcaaataaca agatgacaga cgagtccgag tatggagtta aaactgactc 5520cggatgcccg gagggagcta ggtgttacgt gttcaaccca gaggcagtta acatatccgg 5580gactaaagga gccatggtcc acttacaaaa aactggagga gaattcacct gtgtgacagc 5640atcagggact ccggccttct ttgatctcaa gaacctcaaa ggctggtcag ggctgccgat 5700atttgaggca tcaagtggaa gagtagtcgg cagggttaag gtcgggaaga atgaggactc 5760taaaccaacc aagcttatga gtggaataca aacagtctcc aaaagtacca cagacttgac 5820agaaatggta aagaaaataa caaccatgaa caggggagaa ttcagacaaa taacccttgc 5880cacaggtgcc ggaaaaacca cggaactccc tagatcagtc atagaagaga taggaaggca 5940taagagggtc ttggtcttga tccctctgag ggcggcagca gagtcagtat accaatatat 6000gagacaaaaa cacccaagca tagcattcaa cttgaggata ggggagatga aggaagggga 6060catggccaca gggataacct atgcctcata tggttacttc tgtcagatgc cacaacctaa 6120gctgcgagcc gcgatggttg agtactcctt catattcctt gatgagtacc actgtgccac

6180ccccgaacaa ttggctatca tgggaaagat ccacagattt tcagagaacc tgcgggtagt 6240agccatgacc gcaacaccag caggcacggt aacaactaca gggcaaaaac accctataga 6300agaatacata gccccagaag tgatgaaggg ggaagactta ggttcagagt acttggacat 6360agctggacta aagataccag tagaggagat gaagagtaac atgctggtct ttgtgcccac 6420aaggaacatg gctgtagaga cggcaaagaa actgaaagct aagggttata actcaggcta 6480ctattatagt ggagaggatc catctaacct gagggtggta acatcacagt ccccgtacgt 6540ggtggtagca accaacgcaa tagaatcagg tgttactctc ccagacttgg atgtggtcgt 6600cgacacaggg cttaagtgtg aaaagaggat acggctgtca cctaagatgc ccttcatagt 6660gacgggcctg aagagaatgg ctgtcacgat tggggaacaa gcccagagaa gggggagagt 6720tgggagagtg aagcctggga gatactacag gagtcaagaa acccccgttg gttccaaaga 6780ttaccattac gacctactgc aagcacagag gtacggtata gaagatggga taaacatcac 6840caaatctttt agagagatga attatgattg gagcctttat gaggaggata gtctgatgat 6900tacacaattg gaaatcctca acaatctgtt gatatcagaa gagctaccaa tggcagtaaa 6960aaatataatg gccaggactg accacccaga accaatccaa ctggcgtaca acagctacga 7020aacgcaggtg ccagtgctat tcccaaaaat aaaaaatgga gaggtgactg acagttacga 7080taactatacc ttcctcaacg caagaaagct gggggatgat gtacctccct acgtgtatgc 7140cacagaggat gaggacttag cggtagagct gctgggctta gactggccgg accctgggaa 7200ccaaggaacc gtggaggctg gtagagcact aaaacaagta gttggtctat caacagctga 7260gaacgccctg ttagtagctt tattcggcta tgtaggatat caggcactct caaagaggca 7320tataccagta gtcacagaca tatattcaat tgaagatcac aggttggaag acaccacaca 7380cctacagtat gccccgaatg ctatcaagac ggaggggaag gagacagaat tgaaggagct 7440agctcagggg gatgtgcaga gatgtatgga agctatgact aattatgcaa gagatggcat 7500ccaattcatg aagtctcagg cactgaaagt gaaagaaacc cccacttaca aagagacaat 7560ggacaccgtg gcggactatg taaagaagtt catggaggca ctggcggaca gcaaagaaga 7620catcataaaa tatgggttgt gggggacgca cacaacctta tataagagca tcggtgctag 7680gcttgggaac gagactgcgt tcgctaccct ggtcgtgaaa tggctggcat ttgggggaga 7740atcaatagca gaccatgtca aacaagcggc cacagacttg gtcgtttact atatcatcaa 7800cagacctcag ttcccaggag acacggagac acaacaggaa ggaaggaaat ttgtagccag 7860cctactggtc tcagccctgg ctacttacac ttacaaaagc tggaattaca ataatctgtc 7920caagatagtt gaaccggctt tggctactct gccctatgcc gccacagctc tcaagctatt 7980cgcccccact cgattggaga gcgttgtcat actgagtacc gcaatctaca aaacctacct 8040atcaatcagg cgcggaaaaa gcgatggttt gctaggcaca ggggttagtg cggctatgga 8100aatcatgtca caaaacccag tatctgtggg tatagcggtc atgctagggg tgggggccgt 8160agcggcccac aatgcaatcg aagccagtga gcagaagaga acactactca tgaaagtttt 8220tgtaaagaac ttcttggatc aggcagccac tgatgaatta gtcaaggaga gccctgagaa 8280aataataatg gctttgtttg aagcagtgca gacagtcggc aaccctctta gactggtata 8340ccacctttac ggagtttttt acaaagggtg ggaggcaaaa gagttggccc aaaggacagc 8400cggtaggaat cttttcactt tgataatgtt tgaggctgtg gaactactgg gagtagatag 8460cgaaggaaag atccgccagc tatcaagcaa ttacatacta gagctcctgt ataagttccg 8520tgacagtatc aagtccagcg tgaggcagat ggcaatcagc tgggcccctg ccccttttag 8580ttgtgattgg acaccgacgg atgacagaat agggcttccc caagataatt tcctccgagt 8640ggagacaaaa tgcccctgtg gttacaagat gaaagcagtt aagaattgtg ctggggagtt 8700gagactctta gaagaggaag gctcatttct ctgcaggaat aaattcggga gaggttcacg 8760gaactacagg gtgacaaaat actatgatga caatctatca gaaataaagc cagtgataag 8820aatggaagga catgtggaac tctactacaa gggagccact attaaactgg atttcaacaa 8880cagtaaaaca atattggcaa ccgataaatg ggaggtcgat cactccactc tggtcagggt 8940gctcaagagg cacacagggg ctggatatcg tggggcatac ctgggtgaga aaccgaacca 9000caaacatctg atagagaggg actgcgcaac catcaccaaa gataaggttt gttttctcaa 9060gatgaagaga gggtgtgcat ttacttatga cttatccctt cacaacctta cccggctgat 9120cgaattggta cacaagaata acttggaaga caaagagatt cctgccgtta cggtcacaac 9180ctggctggct tacacatttg taaatgaaga tatagggacc ataaaaccag ccttcgggga 9240gaaaataaca ccagagatgc aggaggagat aaccttgcag cctgctgtag tggtggatgc 9300aactgacgtg accgtgaccg tggtagggga aacccctact atgactacag gggagacccc 9360aacaacgttc accagctcag gtccagaccc gaaaggccaa caagttttaa aactgggagt 9420aggtgaaggc caataccccg ggactaatcc acagagagca agcctgcacg aagccataca 9480aagcgcagat gaaaggccct ctgtgttgat attggggtct gataaagcca cctctaatag 9540agtgaaaact gtaaagaatg tgaaggtata cagaggcagg gacccactag aagtgagaga 9600tatgatgagg aggggaaaga tcctagtcat agccctgtct agggttgata atgctctatt 9660gaaatttgta gattacaaag gcacctttct aactagagag accctggagg cattaagttt 9720gggtaggcca aaaaagaaaa acataaccaa ggcagaagca cagtggttgc tgcgcctcga 9780agaccaaatg gaagagctac ccgattggtt cgcagccggg gaacccattt ttttagaggc 9840caatattaaa catgacaggt atcatctggt aggggatata gctactatca aagagaaagc 9900caaacaattg ggggctacag actctacaaa gatatccaag gaggttggtg caaaagtata 9960ttctatgaaa ttgagtaatt gggtgatgca agaagaaaac aaacagagca acttgacccc 10020cttatttgaa gagctcctac agcagtgtcc acccggaggc caaaacaaaa ctgcacatat 10080ggtctctgct taccaactag ctcaagggaa ctggatgcca accagctgcc atgtttttat 10140ggggaccata tctgccagaa ggactaagac ccatccatat gaagcatatg tcaagttaag 10200ggagttggta gaggaacaca agatgaaaac attgtgtccc ggatcaagtc tgcgtaagca 10260caatgaatgg gtaattggca agatcaaata ccagggcaac ctgaggacca aacacatgtt 10320gaaccccggc aaggtggcag agcaactgca cagagaagga cacagacaca atgtgtataa 10380caagacaata ggctcagtga tgacagctac tggcatcagg ttggagaagt tgcccgtggt 10440tagggcccag acagacacaa ccaacttcca ccaagcaata agggataaga tagacaagga 10500agagaatcta cagaccccgg gtttacataa gaaactaatg gaagttttca atgcattgaa 10560acgacccgag ttagagtcct cctatgacgc tgtggaatgg gaggaattgg agagaggaat 10620aaacagaaag ggtgctgctg gtttctttga acgcaaaaac ataggggaga tattggattc 10680agagaaaaat aaagtagaag agattattga caatctgaaa aagggtagaa atatcaaata 10740ctatgaaacc gcaatcccaa aaaatgaaaa gagggatgtc aatgatgact ggaccgcagg 10800tgactttgtg gacgagaaga aacccagagt catacaatac cctgaagcaa aaacaaggct 10860ggccatcacc aaggtgatgt ataagtgggt gaagcagaag ccagtagtca tacccgggta 10920tgaagggaag acacctctgt tccaaatttt tgacaaagta aagaaggaat gggatcaatt 10980ccaaaatcca gtggcagtga gcttcgacac taaggcgtgg gacacccagg tgaccacaaa 11040tgatctggag ctgataaagg acatacaaaa gtactacttc aagaagaaat ggcataaatt 11100tattgacacc ctgactatgc atatgtcaga agtacccgta atcactgctg atggggaggt 11160gtatataagg aaagggcaaa gaggtagtgg acagcccgac acaagcgcag gcaacagcat 11220gctaaatgtg ttaacaatgg tttatgcctt ctgcgaggcc acaggggtac cctacaagag 11280ttttgacagg gtggcaaaaa ttcatgtgtg cggggacgat ggtttcctga tcacagagag 11340agctctcggc gagaaattcg caagcaaggg agtccaaatc ctgtatgaag ctgggaagcc 11400ccagaagatc actgaagggg acaaaatgaa agtggcctac caatttgatg atattgagtt 11460ttgctcccat acaccaatac aagtaaggtg gtcagataac acttctagct acatgccagg 11520gagaaataca accacaatcc tggctaaaat ggccacaagg ttagattcca gtggtgagag 11580gggtaccata gcgtacgaga aagcagtagc attcagcttc ctgctaatgt attcctggaa 11640cccactaatc agaaggattt gcttattggt actatcaact gaactgcaag tgaaaccagg 11700gaagtcaacc acttactatt atgaagggga cccgatatct gcctacaagg aagtcatcgg 11760ccacaatctt ttcgatctca agagaacaag cttcgagaag ctggccaagt taaatctcag 11820catgtccgta ctcggggcct ggactagaca caccagcaaa agactactac aagactgtgt 11880caatatgggt gttaaagagg gcaactggtt agtcaatgca gacagactgg tgagtagtaa 11940gactggaaat aggtatgtac ctggagaagg ccacaccctg caagggagac attatgaaga 12000actggtgttg gcaagaaaac agatcaacag cttccaaggg acagacaggt acaatctagg 12060cccaatagtc aacatggtgt taaggaggct gagagtcatg atgatgaccc tgatagggag 12120aggggtatga gtgcgggtga cccgcgatct ggacccgtca gtaggaccct attgtagata 12180acactaattt tttatttatt tagatattac tatttattta tttatttatt tattgaatga 12240gtaagaactg gtacaaacta cctcatgtta ccacactaca ctcattttaa cagcacttta 12300gctggaagga aaattcctga cgtccacagt tggactaagg taatttccta acggccc 1235743918PRTClassical Swine Fever Virus 4Met Glu Leu Asn His Phe Glu Leu Leu Tyr Lys Thr Asn Lys Gln Lys 1 5 10 15 Pro Met Gly Val Glu Glu Pro Val Tyr Asp Val Thr Gly Arg Pro Leu 20 25 30 Phe Gly Asp Pro Ser Glu Val His Pro Gln Ser Thr Leu Lys Leu Pro 35 40 45 His Asp Arg Gly Arg Gly Asn Ile Lys Thr Thr Leu Lys Asn Leu Pro 50 55 60 Arg Arg Gly Asp Cys Arg Ser Gly Asn His Leu Gly Pro Val Ser Gly 65 70 75 80 Ile Tyr Val Lys Pro Gly Pro Val Phe Tyr Gln Asp Tyr Met Gly Pro 85 90 95 Val Tyr His Arg Ala Pro Leu Glu Phe Phe Asp Glu Ala Gln Phe Cys 100 105 110 Glu Val Thr Lys Arg Ile Gly Arg Val Thr Gly Ser Asp Gly Lys Leu 115 120 125 Tyr His Ile Tyr Val Cys Ile Asp Gly Cys Ile Leu Leu Lys Leu Ala 130 135 140 Lys Arg Gly Glu Pro Arg Thr Leu Lys Trp Ile Arg Asn Leu Thr Asp 145 150 155 160 Cys Pro Leu Trp Val Thr Ser Cys Ser Asp Asp Gly Ala Ser Ala Ser 165 170 175 Lys Glu Lys Lys Pro Asp Arg Ile Asn Lys Gly Lys Leu Lys Ile Ala 180 185 190 Pro Lys Glu His Glu Lys Asp Ser Arg Thr Lys Pro Pro Asp Ala Thr 195 200 205 Ile Val Val Glu Gly Val Lys Tyr Gln Val Lys Lys Lys Gly Lys Val 210 215 220 Lys Gly Lys Asn Thr Gln Asp Gly Leu Tyr His Asn Lys Asn Lys Pro 225 230 235 240 Pro Glu Ser Arg Lys Lys Leu Glu Lys Ala Leu Leu Ala Trp Ala Val 245 250 255 Ile Ala Ile Met Leu Tyr Gln Pro Val Ala Ala Glu Asn Ile Thr Gln 260 265 270 Trp Asn Leu Ser Asp Asn Gly Thr Asn Gly Ile Gln His Ala Met Tyr 275 280 285 Leu Arg Gly Val Ser Arg Ser Leu His Gly Ile Trp Pro Glu Lys Ile 290 295 300 Cys Lys Gly Val Pro Thr Tyr Leu Ala Thr Asp Thr Glu Leu Arg Glu 305 310 315 320 Ile Gln Gly Met Met Asp Ala Ser Glu Gly Thr Asn Tyr Thr Cys Cys 325 330 335 Lys Leu Gln Arg His Glu Trp Asn Lys His Gly Trp Cys Asn Trp Tyr 340 345 350 Asn Ile Asp Pro Trp Ile Gln Leu Met Asn Arg Thr Gln Ala Asn Leu 355 360 365 Ala Glu Gly Pro Pro Ser Lys Glu Cys Ala Val Thr Cys Arg Tyr Asp 370 375 380 Lys Asn Ala Asp Ile Asn Val Val Thr Gln Ala Arg Asn Arg Pro Thr 385 390 395 400 Thr Leu Thr Gly Cys Lys Lys Gly Lys Asn Phe Ser Phe Ala Gly Thr 405 410 415 Val Ile Glu Gly Pro Cys Asn Phe Asn Val Ser Val Glu Asp Ile Leu 420 425 430 Tyr Gly Asp His Glu Cys Gly Ser Leu Leu Gln Asp Thr Ala Leu Tyr 435 440 445 Leu Val Asp Gly Met Thr Asn Thr Ile Glu Arg Ala Arg Gln Gly Ala 450 455 460 Ala Arg Val Thr Ser Trp Leu Gly Arg Gln Leu Arg Thr Ala Gly Lys 465 470 475 480 Arg Leu Glu Gly Arg Ser Lys Thr Trp Phe Gly Ala Tyr Ala Leu Ser 485 490 495 Pro Tyr Cys Asn Val Thr Ser Lys Ile Gly Tyr Ile Trp Tyr Thr Asn 500 505 510 Asn Cys Thr Pro Ala Cys Leu Pro Lys Asn Thr Lys Ile Ile Gly Pro 515 520 525 Gly Lys Phe Asp Thr Asn Ala Glu Asp Gly Lys Ile Leu His Glu Met 530 535 540 Gly Gly His Leu Ser Glu Phe Leu Leu Leu Ser Leu Val Val Leu Ser 545 550 555 560 Asp Phe Ala Pro Glu Thr Ala Ser Ala Leu Tyr Leu Ile Leu His Tyr 565 570 575 Val Ile Pro Gln Ser His Glu Glu Pro Glu Gly Cys Asp Thr Asn Gln 580 585 590 Leu Asn Leu Thr Val Glu Leu Arg Thr Glu Asp Val Ile Pro Ser Ser 595 600 605 Val Trp Asn Val Gly Lys Tyr Val Cys Val Arg Pro Asp Trp Trp Pro 610 615 620 Tyr Glu Thr Lys Val Ala Leu Leu Phe Glu Glu Ala Gly Gln Val Val 625 630 635 640 Lys Leu Ala Leu Arg Ala Leu Arg Asp Leu Thr Arg Val Trp Asn Ser 645 650 655 Ala Ser Thr Thr Ala Phe Leu Ile Cys Leu Ile Lys Val Leu Arg Gly 660 665 670 Gln Val Val Gln Gly Val Ile Trp Leu Leu Leu Val Thr Gly Ala Pro 675 680 685 Val Ser Cys Thr His Leu Ala Ala Ala Asp Tyr Lys Asp Asp Asp Asp 690 695 700 Lys Gly Ala Gln Gly Arg Leu Ala Cys Lys Glu Asp His Arg Tyr Ala 705 710 715 720 Ile Ser Thr Thr Asn Glu Ile Gly Leu Leu Gly Ala Glu Gly Leu Thr 725 730 735 Thr Thr Trp Lys Glu Tyr Asn His Asn Leu Gln Leu Asp Asp Gly Thr 740 745 750 Val Lys Ala Ile Cys Met Ala Gly Ser Phe Lys Val Thr Ala Leu Asn 755 760 765 Val Val Ser Arg Arg Tyr Leu Ala Ser Leu His Lys Asp Ala Leu Pro 770 775 780 Thr Ser Val Thr Phe Glu Leu Leu Phe Asp Gly Thr Ser Pro Leu Thr 785 790 795 800 Glu Glu Met Gly Asp Asp Phe Gly Phe Gly Leu Cys Pro Tyr Asp Thr 805 810 815 Ser Pro Val Val Lys Gly Lys Tyr Asn Thr Thr Leu Leu Asn Gly Ser 820 825 830 Ala Phe Tyr Leu Val Cys Pro Ile Gly Trp Thr Gly Val Ile Glu Cys 835 840 845 Thr Ser Phe Asn Met Asp Thr Leu Arg Thr Glu Val Val Lys Thr Phe 850 855 860 Arg Arg Glu Lys Pro Phe Pro Tyr Arg Arg Asp Cys Val Thr Thr Thr 865 870 875 880 Val Glu Asn Glu Asp Leu Phe Tyr Cys Lys Trp Gly Gly Asn Trp Thr 885 890 895 Cys Val Lys Gly Glu Pro Val Thr Tyr Thr Gly Gly Pro Val Lys Gln 900 905 910 Cys Arg Trp Cys Gly Phe Asp Phe Asn Glu Pro Asp Gly Leu Pro His 915 920 925 Tyr Pro Ile Gly Lys Cys Ile Leu Ala Asn Glu Thr Gly Tyr Arg Ile 930 935 940 Val Asp Ser Thr Asp Cys Asn Arg Asp Gly Val Val Ile Ser Thr Glu 945 950 955 960 Gly Ser His Glu Cys Leu Ile Gly Asn Thr Thr Val Lys Val His Ala 965 970 975 Leu Asp Glu Arg Leu Gly Pro Met Pro Cys Arg Pro Lys Glu Ile Val 980 985 990 Ser Ser Ala Gly Pro Val Arg Lys Thr Ser Cys Thr Phe Asn Tyr Ala 995 1000 1005 Lys Thr Leu Arg Asn Arg Tyr Tyr Glu Pro Arg Asp Ser Tyr Phe 1010 1015 1020 Gln Gln Tyr Met Leu Lys Gly Glu Tyr Gln Tyr Trp Phe Asp Leu 1025 1030 1035 Asp Val Thr Asp Arg His Ser Asp Tyr Phe Ala Glu Phe Ile Val 1040 1045 1050 Leu Val Val Val Ala Leu Leu Gly Gly Arg Tyr Val Leu Trp Leu 1055 1060 1065 Ile Val Thr Tyr Ile Val Leu Thr Glu Gln Leu Ala Ala Gly Leu 1070 1075 1080 Gln Leu Gly Gln Gly Glu Val Val Leu Ile Gly Asn Leu Ile Thr 1085 1090 1095 His Thr Asp Ile Glu Val Val Val Tyr Phe Leu Leu Leu Tyr Leu 1100 1105 1110 Val Met Arg Asp Glu Pro Ile Lys Lys Trp Ile Leu Leu Leu Phe 1115 1120 1125 His Ala Met Thr Asn Asn Pro Val Lys Thr Ile Thr Val Ala Leu 1130 1135 1140 Leu Met Val Ser Gly Val Ala Lys Gly Gly Lys Ile Asp Gly Gly 1145 1150 1155 Trp Gln Arg Leu Pro Glu Thr Asn Phe Asp Ile Gln Leu Ala Leu 1160 1165 1170 Thr Val Ile Val Val Ala Val Met Leu Leu Ala Lys Lys Asp Pro 1175 1180 1185 Thr Thr Val Pro Leu Val Ile Thr Val Ala Thr Leu Arg Thr Ala 1190 1195 1200 Lys Ile Thr Asn Gly Leu Ser Thr Asp Leu Ala Ile Ala Thr Val 1205 1210 1215 Ser Thr Ala Leu Leu Thr Trp Thr Tyr Ile Ser Asp Tyr Tyr Lys 1220 1225 1230 Tyr Lys Thr Leu Leu Gln Tyr Leu Ile Ser Thr Val Thr Gly Ile 1235 1240 1245 Phe Leu Ile Arg Val Leu Lys Gly Val Gly Glu Leu Asp Leu His 1250 1255 1260 Thr Pro Thr Leu Pro Ser Tyr Arg Pro Leu Phe Phe Ile Leu Val 1265 1270 1275 Tyr Leu Ile Ser Thr Ala Val Val Thr Arg Trp Asn Leu Asp Ile 1280 1285 1290 Ala Gly Leu Leu Leu Gln Cys Val Pro Thr Leu Leu Met Val Phe 1295 1300 1305 Thr Met Trp Ala Asp Ile Leu Thr Leu Ile Leu Ile Leu Pro Thr 1310 1315 1320 Tyr Glu Leu Thr Lys Leu Tyr Tyr Leu Lys Glu Val Lys Ile Gly 1325 1330 1335

Ala Glu Arg Gly Trp Leu Trp Lys Thr Asn Phe Lys Arg Val Asn 1340 1345 1350 Asp Ile Tyr Glu Val Asp Gln Ala Gly Glu Gly Val Tyr Leu Phe 1355 1360 1365 Pro Ser Lys Gln Lys Thr Gly Thr Ile Thr Gly Thr Met Leu Pro 1370 1375 1380 Leu Ile Lys Ala Ile Leu Ile Ser Cys Ile Ser Asn Lys Trp Gln 1385 1390 1395 Phe Ile Tyr Leu Leu Tyr Leu Ile Phe Glu Val Ser Tyr Tyr Leu 1400 1405 1410 His Lys Lys Ile Ile Asp Glu Ile Ala Gly Gly Thr Asn Phe Ile 1415 1420 1425 Ser Arg Leu Val Ala Ala Leu Ile Glu Ala Asn Trp Ala Phe Asp 1430 1435 1440 Asn Glu Glu Val Arg Gly Leu Lys Lys Phe Phe Leu Leu Ser Ser 1445 1450 1455 Arg Val Lys Glu Leu Ile Ile Lys His Lys Val Arg Asn Glu Val 1460 1465 1470 Met Val His Trp Phe Gly Asp Glu Glu Val Tyr Gly Met Pro Lys 1475 1480 1485 Leu Val Gly Leu Val Lys Ala Ala Thr Leu Ser Lys Asn Lys His 1490 1495 1500 Cys Ile Leu Cys Thr Val Cys Glu Asn Arg Glu Trp Arg Gly Glu 1505 1510 1515 Thr Cys Pro Lys Cys Gly Arg Phe Gly Pro Pro Val Thr Cys Gly 1520 1525 1530 Met Thr Leu Ala Asp Phe Glu Glu Lys His Tyr Lys Arg Ile Phe 1535 1540 1545 Phe Arg Glu Asp Gln Ser Glu Gly Pro Val Arg Glu Glu Tyr Ala 1550 1555 1560 Gly Tyr Leu Gln Tyr Arg Ala Arg Gly Gln Leu Phe Leu Arg Asn 1565 1570 1575 Leu Pro Val Leu Ala Thr Lys Val Lys Met Leu Leu Val Gly Asn 1580 1585 1590 Leu Gly Thr Glu Val Gly Asp Leu Glu His Leu Gly Trp Val Leu 1595 1600 1605 Arg Gly Pro Ala Val Cys Lys Lys Val Thr Glu His Glu Lys Cys 1610 1615 1620 Thr Thr Ser Ile Met Asp Lys Leu Thr Ala Phe Phe Gly Val Met 1625 1630 1635 Pro Arg Gly Thr Thr Pro Arg Ala Pro Val Arg Phe Pro Thr Ser 1640 1645 1650 Leu Leu Lys Ile Arg Arg Gly Leu Glu Thr Gly Trp Ala Tyr Thr 1655 1660 1665 His Gln Gly Gly Ile Ser Ser Val Asp His Val Thr Cys Gly Lys 1670 1675 1680 Asp Leu Leu Val Cys Asp Thr Met Gly Arg Thr Arg Val Val Cys 1685 1690 1695 Gln Ser Asn Asn Lys Met Thr Asp Glu Ser Glu Tyr Gly Val Lys 1700 1705 1710 Thr Asp Ser Gly Cys Pro Glu Gly Ala Arg Cys Tyr Val Phe Asn 1715 1720 1725 Pro Glu Ala Val Asn Ile Ser Gly Thr Lys Gly Ala Met Val His 1730 1735 1740 Leu Gln Lys Thr Gly Gly Glu Phe Thr Cys Val Thr Ala Ser Gly 1745 1750 1755 Thr Pro Ala Phe Phe Asp Leu Lys Asn Leu Lys Gly Trp Ser Gly 1760 1765 1770 Leu Pro Ile Phe Glu Ala Ser Ser Gly Arg Val Val Gly Arg Val 1775 1780 1785 Lys Val Gly Lys Asn Glu Asp Ser Lys Pro Thr Lys Leu Met Ser 1790 1795 1800 Gly Ile Gln Thr Val Ser Lys Ser Thr Thr Asp Leu Thr Glu Met 1805 1810 1815 Val Lys Lys Ile Thr Thr Met Asn Arg Gly Glu Phe Arg Gln Ile 1820 1825 1830 Thr Leu Ala Thr Gly Ala Gly Lys Thr Thr Glu Leu Pro Arg Ser 1835 1840 1845 Val Ile Glu Glu Ile Gly Arg His Lys Arg Val Leu Val Leu Ile 1850 1855 1860 Pro Leu Arg Ala Ala Ala Glu Ser Val Tyr Gln Tyr Met Arg Gln 1865 1870 1875 Lys His Pro Ser Ile Ala Phe Asn Leu Arg Ile Gly Glu Met Lys 1880 1885 1890 Glu Gly Asp Met Ala Thr Gly Ile Thr Tyr Ala Ser Tyr Gly Tyr 1895 1900 1905 Phe Cys Gln Met Pro Gln Pro Lys Leu Arg Ala Ala Met Val Glu 1910 1915 1920 Tyr Ser Phe Ile Phe Leu Asp Glu Tyr His Cys Ala Thr Pro Glu 1925 1930 1935 Gln Leu Ala Ile Met Gly Lys Ile His Arg Phe Ser Glu Asn Leu 1940 1945 1950 Arg Val Val Ala Met Thr Ala Thr Pro Ala Gly Thr Val Thr Thr 1955 1960 1965 Thr Gly Gln Lys His Pro Ile Glu Glu Tyr Ile Ala Pro Glu Val 1970 1975 1980 Met Lys Gly Glu Asp Leu Gly Ser Glu Tyr Leu Asp Ile Ala Gly 1985 1990 1995 Leu Lys Ile Pro Val Glu Glu Met Lys Ser Asn Met Leu Val Phe 2000 2005 2010 Val Pro Thr Arg Asn Met Ala Val Glu Thr Ala Lys Lys Leu Lys 2015 2020 2025 Ala Lys Gly Tyr Asn Ser Gly Tyr Tyr Tyr Ser Gly Glu Asp Pro 2030 2035 2040 Ser Asn Leu Arg Val Val Thr Ser Gln Ser Pro Tyr Val Val Val 2045 2050 2055 Ala Thr Asn Ala Ile Glu Ser Gly Val Thr Leu Pro Asp Leu Asp 2060 2065 2070 Val Val Val Asp Thr Gly Leu Lys Cys Glu Lys Arg Ile Arg Leu 2075 2080 2085 Ser Pro Lys Met Pro Phe Ile Val Thr Gly Leu Lys Arg Met Ala 2090 2095 2100 Val Thr Ile Gly Glu Gln Ala Gln Arg Arg Gly Arg Val Gly Arg 2105 2110 2115 Val Lys Pro Gly Arg Tyr Tyr Arg Ser Gln Glu Thr Pro Val Gly 2120 2125 2130 Ser Lys Asp Tyr His Tyr Asp Leu Leu Gln Ala Gln Arg Tyr Gly 2135 2140 2145 Ile Glu Asp Gly Ile Asn Ile Thr Lys Ser Phe Arg Glu Met Asn 2150 2155 2160 Tyr Asp Trp Ser Leu Tyr Glu Glu Asp Ser Leu Met Ile Thr Gln 2165 2170 2175 Leu Glu Ile Leu Asn Asn Leu Leu Ile Ser Glu Glu Leu Pro Met 2180 2185 2190 Ala Val Lys Asn Ile Met Ala Arg Thr Asp His Pro Glu Pro Ile 2195 2200 2205 Gln Leu Ala Tyr Asn Ser Tyr Glu Thr Gln Val Pro Val Leu Phe 2210 2215 2220 Pro Lys Ile Lys Asn Gly Glu Val Thr Asp Ser Tyr Asp Asn Tyr 2225 2230 2235 Thr Phe Leu Asn Ala Arg Lys Leu Gly Asp Asp Val Pro Pro Tyr 2240 2245 2250 Val Tyr Ala Thr Glu Asp Glu Asp Leu Ala Val Glu Leu Leu Gly 2255 2260 2265 Leu Asp Trp Pro Asp Pro Gly Asn Gln Gly Thr Val Glu Ala Gly 2270 2275 2280 Arg Ala Leu Lys Gln Val Val Gly Leu Ser Thr Ala Glu Asn Ala 2285 2290 2295 Leu Leu Val Ala Leu Phe Gly Tyr Val Gly Tyr Gln Ala Leu Ser 2300 2305 2310 Lys Arg His Ile Pro Val Val Thr Asp Ile Tyr Ser Ile Glu Asp 2315 2320 2325 His Arg Leu Glu Asp Thr Thr His Leu Gln Tyr Ala Pro Asn Ala 2330 2335 2340 Ile Lys Thr Glu Gly Lys Glu Thr Glu Leu Lys Glu Leu Ala Gln 2345 2350 2355 Gly Asp Val Gln Arg Cys Met Glu Ala Met Thr Asn Tyr Ala Arg 2360 2365 2370 Asp Gly Ile Gln Phe Met Lys Ser Gln Ala Leu Lys Val Lys Glu 2375 2380 2385 Thr Pro Thr Tyr Lys Glu Thr Met Asp Thr Val Ala Asp Tyr Val 2390 2395 2400 Lys Lys Phe Met Glu Ala Leu Ala Asp Ser Lys Glu Asp Ile Ile 2405 2410 2415 Lys Tyr Gly Leu Trp Gly Thr His Thr Thr Leu Tyr Lys Ser Ile 2420 2425 2430 Gly Ala Arg Leu Gly Asn Glu Thr Ala Phe Ala Thr Leu Val Val 2435 2440 2445 Lys Trp Leu Ala Phe Gly Gly Glu Ser Ile Ala Asp His Val Lys 2450 2455 2460 Gln Ala Ala Thr Asp Leu Val Val Tyr Tyr Ile Ile Asn Arg Pro 2465 2470 2475 Gln Phe Pro Gly Asp Thr Glu Thr Gln Gln Glu Gly Arg Lys Phe 2480 2485 2490 Val Ala Ser Leu Leu Val Ser Ala Leu Ala Thr Tyr Thr Tyr Lys 2495 2500 2505 Ser Trp Asn Tyr Asn Asn Leu Ser Lys Ile Val Glu Pro Ala Leu 2510 2515 2520 Ala Thr Leu Pro Tyr Ala Ala Thr Ala Leu Lys Leu Phe Ala Pro 2525 2530 2535 Thr Arg Leu Glu Ser Val Val Ile Leu Ser Thr Ala Ile Tyr Lys 2540 2545 2550 Thr Tyr Leu Ser Ile Arg Arg Gly Lys Ser Asp Gly Leu Leu Gly 2555 2560 2565 Thr Gly Val Ser Ala Ala Met Glu Ile Met Ser Gln Asn Pro Val 2570 2575 2580 Ser Val Gly Ile Ala Val Met Leu Gly Val Gly Ala Val Ala Ala 2585 2590 2595 His Asn Ala Ile Glu Ala Ser Glu Gln Lys Arg Thr Leu Leu Met 2600 2605 2610 Lys Val Phe Val Lys Asn Phe Leu Asp Gln Ala Ala Thr Asp Glu 2615 2620 2625 Leu Val Lys Glu Ser Pro Glu Lys Ile Ile Met Ala Leu Phe Glu 2630 2635 2640 Ala Val Gln Thr Val Gly Asn Pro Leu Arg Leu Val Tyr His Leu 2645 2650 2655 Tyr Gly Val Phe Tyr Lys Gly Trp Glu Ala Lys Glu Leu Ala Gln 2660 2665 2670 Arg Thr Ala Gly Arg Asn Leu Phe Thr Leu Ile Met Phe Glu Ala 2675 2680 2685 Val Glu Leu Leu Gly Val Asp Ser Glu Gly Lys Ile Arg Gln Leu 2690 2695 2700 Ser Ser Asn Tyr Ile Leu Glu Leu Leu Tyr Lys Phe Arg Asp Ser 2705 2710 2715 Ile Lys Ser Ser Val Arg Gln Met Ala Ile Ser Trp Ala Pro Ala 2720 2725 2730 Pro Phe Ser Cys Asp Trp Thr Pro Thr Asp Asp Arg Ile Gly Leu 2735 2740 2745 Pro Gln Asp Asn Phe Leu Arg Val Glu Thr Lys Cys Pro Cys Gly 2750 2755 2760 Tyr Lys Met Lys Ala Val Lys Asn Cys Ala Gly Glu Leu Arg Leu 2765 2770 2775 Leu Glu Glu Glu Gly Ser Phe Leu Cys Arg Asn Lys Phe Gly Arg 2780 2785 2790 Gly Ser Arg Asn Tyr Arg Val Thr Lys Tyr Tyr Asp Asp Asn Leu 2795 2800 2805 Ser Glu Ile Lys Pro Val Ile Arg Met Glu Gly His Val Glu Leu 2810 2815 2820 Tyr Tyr Lys Gly Ala Thr Ile Lys Leu Asp Phe Asn Asn Ser Lys 2825 2830 2835 Thr Ile Leu Ala Thr Asp Lys Trp Glu Val Asp His Ser Thr Leu 2840 2845 2850 Val Arg Val Leu Lys Arg His Thr Gly Ala Gly Tyr Arg Gly Ala 2855 2860 2865 Tyr Leu Gly Glu Lys Pro Asn His Lys His Leu Ile Glu Arg Asp 2870 2875 2880 Cys Ala Thr Ile Thr Lys Asp Lys Val Cys Phe Leu Lys Met Lys 2885 2890 2895 Arg Gly Cys Ala Phe Thr Tyr Asp Leu Ser Leu His Asn Leu Thr 2900 2905 2910 Arg Leu Ile Glu Leu Val His Lys Asn Asn Leu Glu Asp Lys Glu 2915 2920 2925 Ile Pro Ala Val Thr Val Thr Thr Trp Leu Ala Tyr Thr Phe Val 2930 2935 2940 Asn Glu Asp Ile Gly Thr Ile Lys Pro Ala Phe Gly Glu Lys Ile 2945 2950 2955 Thr Pro Glu Met Gln Glu Glu Ile Thr Leu Gln Pro Ala Val Val 2960 2965 2970 Val Asp Ala Thr Asp Val Thr Val Thr Val Val Gly Glu Thr Pro 2975 2980 2985 Thr Met Thr Thr Gly Glu Thr Pro Thr Thr Phe Thr Ser Ser Gly 2990 2995 3000 Pro Asp Pro Lys Gly Gln Gln Val Leu Lys Leu Gly Val Gly Glu 3005 3010 3015 Gly Gln Tyr Pro Gly Thr Asn Pro Gln Arg Ala Ser Leu His Glu 3020 3025 3030 Ala Ile Gln Ser Ala Asp Glu Arg Pro Ser Val Leu Ile Leu Gly 3035 3040 3045 Ser Asp Lys Ala Thr Ser Asn Arg Val Lys Thr Val Lys Asn Val 3050 3055 3060 Lys Val Tyr Arg Gly Arg Asp Pro Leu Glu Val Arg Asp Met Met 3065 3070 3075 Arg Arg Gly Lys Ile Leu Val Ile Ala Leu Ser Arg Val Asp Asn 3080 3085 3090 Ala Leu Leu Lys Phe Val Asp Tyr Lys Gly Thr Phe Leu Thr Arg 3095 3100 3105 Glu Thr Leu Glu Ala Leu Ser Leu Gly Arg Pro Lys Lys Lys Asn 3110 3115 3120 Ile Thr Lys Ala Glu Ala Gln Trp Leu Leu Arg Leu Glu Asp Gln 3125 3130 3135 Met Glu Glu Leu Pro Asp Trp Phe Ala Ala Gly Glu Pro Ile Phe 3140 3145 3150 Leu Glu Ala Asn Ile Lys His Asp Arg Tyr His Leu Val Gly Asp 3155 3160 3165 Ile Ala Thr Ile Lys Glu Lys Ala Lys Gln Leu Gly Ala Thr Asp 3170 3175 3180 Ser Thr Lys Ile Ser Lys Glu Val Gly Ala Lys Val Tyr Ser Met 3185 3190 3195 Lys Leu Ser Asn Trp Val Met Gln Glu Glu Asn Lys Gln Ser Asn 3200 3205 3210 Leu Thr Pro Leu Phe Glu Glu Leu Leu Gln Gln Cys Pro Pro Gly 3215 3220 3225 Gly Gln Asn Lys Thr Ala His Met Val Ser Ala Tyr Gln Leu Ala 3230 3235 3240 Gln Gly Asn Trp Met Pro Thr Ser Cys His Val Phe Met Gly Thr 3245 3250 3255 Ile Ser Ala Arg Arg Thr Lys Thr His Pro Tyr Glu Ala Tyr Val 3260 3265 3270 Lys Leu Arg Glu Leu Val Glu Glu His Lys Met Lys Thr Leu Cys 3275 3280 3285 Pro Gly Ser Ser Leu Arg Lys His Asn Glu Trp Val Ile Gly Lys 3290 3295 3300 Ile Lys Tyr Gln Gly Asn Leu Arg Thr Lys His Met Leu Asn Pro 3305 3310 3315 Gly Lys Val Ala Glu Gln Leu His Arg Glu Gly His Arg His Asn 3320 3325 3330 Val Tyr Asn Lys Thr Ile Gly Ser Val Met Thr Ala Thr Gly Ile 3335 3340 3345 Arg Leu Glu Lys Leu Pro Val Val Arg Ala Gln Thr Asp Thr Thr 3350 3355 3360 Asn Phe His Gln Ala Ile Arg Asp Lys Ile Asp Lys Glu Glu Asn 3365 3370 3375 Leu Gln Thr Pro Gly Leu His Lys Lys Leu Met Glu Val Phe Asn 3380 3385 3390 Ala Leu Lys Arg Pro Glu Leu Glu Ser Ser Tyr Asp Ala Val Glu 3395 3400 3405 Trp Glu Glu Leu Glu Arg Gly Ile Asn Arg Lys Gly Ala Ala Gly 3410 3415 3420 Phe Phe Glu Arg Lys Asn Ile Gly Glu Ile Leu Asp Ser Glu Lys 3425 3430 3435 Asn Lys Val Glu Glu Ile Ile Asp Asn Leu Lys Lys Gly Arg Asn 3440 3445 3450 Ile Lys Tyr Tyr Glu Thr Ala Ile Pro Lys Asn Glu Lys Arg Asp 3455 3460 3465 Val Asn Asp Asp Trp Thr Ala Gly Asp Phe Val Asp Glu Lys Lys 3470 3475 3480 Pro Arg Val Ile Gln Tyr Pro Glu Ala Lys Thr Arg Leu Ala Ile 3485 3490 3495 Thr Lys Val Met Tyr Lys Trp Val Lys Gln Lys Pro Val Val Ile 3500 3505 3510 Pro Gly Tyr Glu Gly Lys Thr Pro Leu Phe Gln Ile Phe Asp Lys 3515 3520 3525 Val Lys Lys Glu Trp Asp Gln Phe Gln Asn

Pro Val Ala Val Ser 3530 3535 3540 Phe Asp Thr Lys Ala Trp Asp Thr Gln Val Thr Thr Asn Asp Leu 3545 3550 3555 Glu Leu Ile Lys Asp Ile Gln Lys Tyr Tyr Phe Lys Lys Lys Trp 3560 3565 3570 His Lys Phe Ile Asp Thr Leu Thr Met His Met Ser Glu Val Pro 3575 3580 3585 Val Ile Thr Ala Asp Gly Glu Val Tyr Ile Arg Lys Gly Gln Arg 3590 3595 3600 Gly Ser Gly Gln Pro Asp Thr Ser Ala Gly Asn Ser Met Leu Asn 3605 3610 3615 Val Leu Thr Met Val Tyr Ala Phe Cys Glu Ala Thr Gly Val Pro 3620 3625 3630 Tyr Lys Ser Phe Asp Arg Val Ala Lys Ile His Val Cys Gly Asp 3635 3640 3645 Asp Gly Phe Leu Ile Thr Glu Arg Ala Leu Gly Glu Lys Phe Ala 3650 3655 3660 Ser Lys Gly Val Gln Ile Leu Tyr Glu Ala Gly Lys Pro Gln Lys 3665 3670 3675 Ile Thr Glu Gly Asp Lys Met Lys Val Ala Tyr Gln Phe Asp Asp 3680 3685 3690 Ile Glu Phe Cys Ser His Thr Pro Ile Gln Val Arg Trp Ser Asp 3695 3700 3705 Asn Thr Ser Ser Tyr Met Pro Gly Arg Asn Thr Thr Thr Ile Leu 3710 3715 3720 Ala Lys Met Ala Thr Arg Leu Asp Ser Ser Gly Glu Arg Gly Thr 3725 3730 3735 Ile Ala Tyr Glu Lys Ala Val Ala Phe Ser Phe Leu Leu Met Tyr 3740 3745 3750 Ser Trp Asn Pro Leu Ile Arg Arg Ile Cys Leu Leu Val Leu Ser 3755 3760 3765 Thr Glu Leu Gln Val Lys Pro Gly Lys Ser Thr Thr Tyr Tyr Tyr 3770 3775 3780 Glu Gly Asp Pro Ile Ser Ala Tyr Lys Glu Val Ile Gly His Asn 3785 3790 3795 Leu Phe Asp Leu Lys Arg Thr Ser Phe Glu Lys Leu Ala Lys Leu 3800 3805 3810 Asn Leu Ser Met Ser Val Leu Gly Ala Trp Thr Arg His Thr Ser 3815 3820 3825 Lys Arg Leu Leu Gln Asp Cys Val Asn Met Gly Val Lys Glu Gly 3830 3835 3840 Asn Trp Leu Val Asn Ala Asp Arg Leu Val Ser Ser Lys Thr Gly 3845 3850 3855 Asn Arg Tyr Val Pro Gly Glu Gly His Thr Leu Gln Gly Arg His 3860 3865 3870 Tyr Glu Glu Leu Val Leu Ala Arg Lys Gln Ile Asn Ser Phe Gln 3875 3880 3885 Gly Thr Asp Arg Tyr Asn Leu Gly Pro Ile Val Asn Met Val Leu 3890 3895 3900 Arg Arg Leu Arg Val Met Met Met Thr Leu Ile Gly Arg Gly Val 3905 3910 3915 554DNAClassical Swine Fever Virus 5 ccagtgagtt gcacccactt agccgcggcc gactataaag acgatgatga caaa 54654DNAClassical Swine Fever Virus 6cctgtctctt gtacacatct tgcggccgca gattacaagg atgacgacga taag 54718PRTClassical Swine Fever Virus 7Pro Val Ser Cys Thr His Leu Ala Ala Ala Asp Tyr Lys Asp Asp Asp 1 5 10 15 Asp Lys 858DNAArtificial SequenceChemically Synthesized 8gttactggta actggggcac cagtgagttg cacccacctt gcggccgcag attacaag 58958DNAArtificial SequenceChemically Synthesized 9cttgtaatct gcggccgcaa ggtgggtgca actcactggt gccccagtta ccagtaac 581058DNAArtificial SequenceChemically Synthesized 10caccagtgag ttgcacccac ttagccgcgg ccgactataa ggatgacgac gataagca 581158DNAArtificial SequenceChemically Synthesized 11tgcttatcgt cgtcatcctt atagtcggcc gcggctaagt gggtgcaact cactggtg 581255DNAArtificial SequenceChemically Synthesized 12cccacttagc cgcggccgac tataaagacg atgatgacaa acaaggccgg ctagc 551355DNAArtificial SequenceChemically Synthesized 13gctagccggc cttgtttgtc atcatcgtct ttatagtcgg ccgcggctaa gtggg 551415DNAClassical Swine Fever Virus 14tcatttaata tggac 15155PRTClassical Swine Fever Virus 15Ser Phe Asn Met Asp 1 5 1615DNAClassical Swine Fever Virus 16agtttcggaa tggat 15175PRTClassical Swine Fever Virus 17Ser Phe Gly Met Asp 1 5 1846DNAArtificial SequenceChemically Synthesized 18gacgggtgtt atagagtgca cgagtttcgg aatggatact ctgaga 461948DNAArtificial SequenceChemically Synthesized 19tctcagagta tccttccgaa acttgacgtg cactctataa cacccgtc 48