High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus.
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.
Shenyang
Gao
G
Veterinary Department, Northeast Agricultural University, Harbin 150030, PR China.
Enhui
Zha
Z
Baoxian
Li
L
Xinyuan
Qiao
Q
Lijie
Tang
T
Junwei
Ge
G
Yijing
Li
L
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
D000086862:Coronavirus M Proteins; C000706236:M protein, Porcine epidemic diarrhea virus; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins