Preparation Of Bovine Herpesvirus-1 Recombinant Antigen And Development Of Indirect ELISA

Infectious bovine rhniotracheitis (IBR), which was caused by Bovine herpesvirus-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. In this study, the gB and gE gene were cloned, the prokaryotic and eukaryotic expression vectors were constructed, the high efficient expression of gB was obtained, and the Indirect ELISA to detect BHV-1 was established.Firstly, the antigenic analysis was made to gB,gE protein , and according to its immunogenicity and the demands of the vectors , several primers were designed. The gB,gE genes all-length encoding sequences were amplified by PCR, then gB and gE genes were cloned into pGEM-T-easy. With the recombinant plasmids pGEM-T-gB and pGEM-T-gE as the template, the gB and gE fragments were amplified through the PCR, then the fragments were inserted into the vectors: pET-32a, and ultimately the prokaryotic expression systems with BL21(DE3) as the host were conducted. Through the studies on the induction concentration and time of IPTG and the culture time of hosts, it was established that the gBâ… , gBâ…¡and gBâ…¢, which encode the gB 28~216 aa, gB 236~418 aa and gB 384~639 aa, could be expressed in pET-32a/BL21(DE3). The products of...