earticle

영어

Porcine epidemic diarrhea virus (PEDV) is a single stranded RNA virus that belongs to the Coronaviridae family. The envelope contains three main viral proteins, spike (S, 180–20 kDa), membrane (M, 27–2 kDa and small membrane(sM, 7 kDa). The M protein is the most abundant of three; it is a triple-spanning membrane protein with a short amino-terminal domain on the outside of the virus and a long carboxyterminal domain on the inside. In order to obtain and assess the recombinant M protein used for a diagnostic reagent of porcine epidemic diarrhea (PED). M protein gene of KPEDV-9 strain was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR). Then, the M protein gene was cloned into prokaryotic expression vector pET-21a. The recombinant plasmid pET-21a-M was constructed and transformed into E. coli BL21(DE3) for expression. The M protein gene of KPEDV-9 strain consisted of 692 nucleotides containing a single open reading frame (ORF) of 681 nucleotides, which encoded a 226aa-long peptide. SDS-PAGE analysis showed recombinant M protein was highly expressed by pET-21a-M and the product fusion protein his-tag-M reached 40% in the total bacteria proteins. The preliminary purified recombinant protein was evaluated for its antigenicity and reactivity through western blotting. The results indicated the recombinant M protein should be candidate as a feasible recombinant diagnostic reagent for preventing and control of PED.