Peer-reviewed veterinary case report
High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus.
- Journal:
- Veterinary microbiology
- Year:
- 2007
- Authors:
- Shenyang, Gao et al.
- Affiliation:
- Veterinary Department · China
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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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/17475420/