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  • Title: High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus.
    Author: Shenyang G, Enhui Z, Baoxian L, Xinyuan Q, Lijie T, Junwei G, Yijing L.
    Journal: Vet Microbiol; 2007 Jul 20; 123(1-3):187-93. PubMed ID: 17475420.
    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.
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