Authors : Jianke Wang, Qiang Zhang, Lei Wu and Wenhui Wang
Abstract: The P12A gene fragments and 3C gene fragments of O/China 99 strains of foot and mouth virus were ligated into bilateral tandem Vaccinia virus promoter P7.5 which could initiate expression of EGFP gene and the expression box p-EGFP-N1-P7.5-P12A-3C was constructed. The expression box was ligated to the linear vector pUC119-TK after KpnI enzyme digestion by blunt end ligation to obtain the recombinant plasmid pUC119-TK-EGFP-P7.5-P12A-3C. The homologous recombination took place in the BHK-21 cell between the recombinant plasmid and the Capripox virus attenuated strain by deleted TK gene. The PCR identification, the Western blot analysis and immunological experiment on mice were carried out. The results showed that the recombinant strain could be subcultured in BHK-21 cell for 10 passages, expressing P12A-3C gene which was 2900 bp by sequence analysis, the Western blot analysis demonstrated that the expressed protein of recombinant plasmid pUC119-TK-EGFP-N1-P7.5-P12A-3C in BHK-21 cells infected GTPV AV41 could be specifically recognized by the hyperimmune serum to type O FMDV. Immunity assay showed that the expression protein had immunogenicity in mice. All the results showed that the Goat Pox virus attenuated strain which could express P12A-3C gene of type O FMDV was obtained.
Jianke Wang, Qiang Zhang, Lei Wu and Wenhui Wang, 2013. Expression and Study on Immunogenicity of P12A-3A Genes of Type O Foot and Mouth Disease Virus in Recombinant Attenuated Goatpox Virus Strain. Journal of Animal and Veterinary Advances, 12: 1059-1065.