Prokaryotic Expression Of GC And GD Gene Of IBRV And Construction Of Eukaryotic Expression Vector

Infectious bovine rhinotracheitis(IBR), which was caused by the Bovine herpersvirus-1(BHV-1), was acute, pyrexic and contagious, and incurred great economic loss to the global cattle husbandry. IBR was listed as the B disease by OIE, and was the main object quarantined in the international trade and the animals imported to China.Glycoprotein D was the major structure protein and the protective antigen, and it plays an important role in the pathogenic process and immune response. Glycoprotein C could induce important biofunctions and induce cellular immune response. In the studies, The gC and gD genes of IBRV Bartha Nu/67 strain were amplified by PCR, and were cloned into the E.coli. The eukaryotic expression vector Bacmid was constructed, which made the base for the transfection of the Sf9 cells.1. Two primers were designed and synthesized according to the gene sequence of IBRV Bartha Nu/67 strain in the Genbank, and the gC and gD genes were amplified by PCR, cloned and sequenced. The sequence analysis of the product showed that the gC and gD gene had been successfully obtained, which were 1311bp and 1251bp, respectively. The sequence comparison analysis confirmed that the only one base of gD gene was different form the standard, the homogeneity was 99.82% and the amino acid encoded by the gene was completely the same as the standard, wghich meant the mutation of the base could not induce the change of amino acid. The homogeneity of gC gene product and the amino acid encoded by gC was100%, which confirmed the highly conservation of IBRV gene. According to the gene analysis, four pairs primers with the enzyme scission site were designed. The prokaryotic expression fragments were amplified from the pGEM T-gCå’ŒpGEM T-gD...