title: 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.
authors.affiliation: Veterinary Department, Northeast Agricultural University, Harbin 150030, PR China.
authors.forename: Gao
authors.initials: G
authors.lastname: Shenyang
authors.forename: Zha
authors.initials: Z
authors.lastname: Enhui
authors.forename: Li
authors.initials: L
authors.lastname: Baoxian
authors.forename: Qiao
authors.initials: Q
authors.lastname: Xinyuan
authors.forename: Tang
authors.initials: T
authors.lastname: Lijie
authors.forename: Ge
authors.initials: G
authors.lastname: Junwei
authors.forename: Li
authors.initials: L
authors.lastname: Yijing
mesh_terms: D000818:Animals; D018352:Coronavirus Infections / Q000662:veterinary / Q000821:virology; D000086862:Coronavirus M Proteins; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D051379:Mice; D008807:Mice, Inbred BALB C; D053485:Porcine epidemic diarrhea virus / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000821:virology; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000086862:Coronavirus M Proteins; C000706236:M protein, Porcine epidemic diarrhea virus; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins