Expression and Antigenic Characterization of the Epitope-G1 of Bovine Ephemeral Fever Virus Glycoprotein in Pichia pastoris

Abstract:

The epitope-G₁ gene, cloned from the pMD-G plasmid including G protein gene of bovine ephemeral fever virus, was subcloned into expression vector pPIC9K to construct pPIC9K-G₁ recombinant plasmid successfully, and the recombinant plasmid linearized was transformed into Pichia pastoris GS115 by electroporation. The recombinant Pichia pastoris strains were screened by G418 and PCR, and induced by methanol. The expressed products were analyzed by SDS-PAGE, deglycosylation, Western blot, ELISA, immunizing rabbits and specificity experiments. The results indicated that the gene was expressed successfully in GS115 and glycosylated moderately, and the target protein had nicer biological activity and specificity. The protein is able to be used as coating antigen to develop ELISA Kit for diagnosing bovine ephemeral fever.

Key words:Bovine ephemeral fever virus, Epitope-G₁, Pichia pastoris, Expression, Characterization