Studies On Goatpox Virus Metabolomics And Construction Of Live Vector Vaccine Expressing Epitopes Of Foot-and-mouth Disease Virus

Capripox is an acute feverish and highly contagious disease which caused by sheep pox virus infecting sheep or goat pox virus infecting goat, it is the only double stranded DNA and capsulated virus copying in the cytoplasm. Both FMD and CP can cause serious harm, causing huge economic losses, are hazardousextremely serious disease were classified as group A diseases and must be reported to Office International Des Epizooties(OIE) while occured. Foot-and-mouth disease(FMD), caused major animal disease of cattle, sheep and pigs and other economically important livestock by Foot-and-mouth disease virus disease(FMDV). FMDV host range widely and spread fast, and more serotypes, respectively, O, A, C, Asial, SAT1, SAT2 and SAT3 seven serotypes.In this study, FMDV type O B-cell epitopes were screening and identificated. According the principle of homologous recombination an epitope expression cassette containing foreign gene is constructed. Then the expression cassette is inserted into the vector which comprising Thymidine kinase gene and it is the left and right homology arms.The transfer vector plasmid is constructed and obtained. The transfer vector transfected cells which infected goat-pox virus attenuated vaccine strain, screening recombinant viruses.(1)Screening and identification of FMDV O/Mya98 strain B-cell epitopes.The purpose of this study was to screening and identification B-cell epitopes of FMDV O/Mya98 strain. DNASTAR tool was used to analyze the four structural proteins (VP1, VP2, VP3, and VP4) complete amino acid sequences of FMDV. The hydrophilicity, flexibility, antigenic index, and surface probability of the P1 structural protein of FMDV were analyzed using the Kyte-Doolittle program, the Karplus-Schulz program, the Jameson-Wolf program, and the Emini program, respectively. The above indicators were summarized, and the results showed that the potential B-cell epitopes of the P1 structural protein were β turns with strong hydrophilicity and high flexibility or irregular coils. Epitopes is identified by Western blot and ELISA.(2) Construct recombinant carpripox virus expression FMDV serotype O epitopesAccording to the principle of homologous recombination, the promoter (P7.5/ 11), the reporter gene (EGFP and gpt), the target gene epitope (Epitope), regulatory elements, terminator were cloned and connected to the T-vector, then digested series to form gene expression cassette; TK gene is selected as the extraneous gene cloning site, the gene extraction of goat-pox virus strains AV41, cloned and sequenced homology arm containing the TK gene ORF and its both sides connected to T-vector; The expression cassette contains elements of each reaction was inserted into PGEM-TK obtained a transfer vector named P7.5-EGFP-Pll-gpt-Epitope; It transfected into cells.(3) Preliminary screening of recombinant virusesMetabonomics analysis is performed for the infected and uninfected cell with goat-pox virus. To provide basis for the metabonomics analysis of sheep testicular cells infected with recombinant virus and to establish a method for the selection of recombinant viruses.