Expression of G۱- epitope of bovine ephemeral fever virus in E. coli : A novel candidate to develop ELISA kit

Bovine ephemeral fever is an acute and arthropod-borne viral disease of cattle and water buffalo which occurs seasonally in most of the world tropical and subtropical regions. The epizootic feature of the disease has been reported in Iran with serious economic consequences. The surface glycoprotein G of bovine ephemeral fever virus (BEFV) is composed of ۴ antigenic sites (G۱-G۴) and plays the main role for eliciting neutralizing antibodies and protective immunity. The G۱ – epitope is a linear antigenic site and conserved among BEFV strains. In order to develop an ELISA test based on G۱-epitope as coating antigen, this study was carried out to express the recombinant G۱-epitope of BEFV in prokaryotic system. Using PCR and specific primers, a length of ۸۸ amino acid of the G glycoprotein of BEFV including G۱- epitope was amplified and cloned into the expression vector pGEX-۴T-۱, with the GST moiety. The recombinant plasmid (pGEX-۴T-۱-G۱) was then transformed into Escherichia coli BL۲۱ and expression of fusion protein was induced by ۰.۱۰ mM IPTG. The maximum expression of the fusion protein was obtained at ۱۶ hr post induction as verified by SDS-PAGE electrophoresis, and it was also confirmed that this protein bearing G۱- epitope is sufficiently biologically active to bind to anti-BEFV serum in western blot experiment.

کلیدواژه ها:

BEFV ، Escherichia coli ، ELISA ، G۱- Epitope