Gao Shenyang

High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus

Abstract The truncated fragment M′ gene, encoding the exterior of the viral envelope protein of PEDV, was subcloned into prokaryotic expression vector pGEX-6p-1. The recombinant plasmid pGEX-6p-M′ was constructed and transformed into E. coli BL21(DE3)pLysS for expression. SDS-PAGE analysis showed recombinant truncated M′ protein was highly expressed by pGEX-6p-M′ and the product fusion protein GST-M′ reached 45% in the total bacteria proteins with the analysis of software AlphaImager2200. The preliminary purified recombinant protein was evaluated for its antigenicity and reactivity through Western blotting and indirect enzyme-linked immunosorbent assay (ELISA) with monoclonal antibody against M protein of PEDV and porcine polyclonal anti-PEDV antiserum as the primary antibody. The results indicated the recombinant truncated M′ protein should be candidate as a feasible recombinant diagnostic reagent.