Gene Cloning and Expression of Ostreid Herpesvirus 1 Envelope Protein (ORF111)

Abstract:

The envelope protein (ORF111) of Ostreid Herpesvirus 1 (OsHV-1) were studied using bioinformatics analysis, gene cloning and expression. Firstly, we designed the primers of the orf111 gene based on the complete genome sequence of OsHV-1, and successfully cloned the gene. We further analyzed the biological characteristics of the deduced protein sequence encoded by ORF111 gene, which including physicochemical properties, advanced structure, transmembrane region, and antigen determinant cluster. The results showed that the ORF111 protein is a stable hydrophobic protein, which contains nine antigenic determinants, five transmembrane domain structures, and a highly conservative pure ammonia arginyl-glycyl-aspartate (Arg-Gly-Asp, RGD) sequence. Then the OsHV-1-orf111 gene was linked with a pET28a(+) plasmid, and the recombinant plasmid pET28a-orf111 was successfully constructed. Finally, the recombinant plasmid was transformed into DH5α, and the recombinant protein was expressed after inducing by isopropyl-beta-D-thiogalactosine (IPTG). The SDS-PAGE test showed that the molecular weight of the ORF111 protein was about 32 kDa. In this study, an OsHV-1 envelope protein containing the RGD domain was successfully obtained by prokaryotic expression, which lays a foundation for the preparation of monoclonal antibodies of ORF111, and provides important basis for further study of the infection mechanism of OsHV-1. It also stimulates new ideas for the prevention and control of OsHV-1.