earticle

영어

Porcine epidemic diarrhea virus (PEDV) causes a devastating enteric disease with acute diarrhea, dehydration and significant mortality in swine. Because of the high mortality which is up to 100% in suckling piglets, PED is an important porcine disease in Korea. PEDV, a member of the family Coronaviridae, is an enveloped virus possessing a singlestranded positive-sense approximately 28 kb RNA genome with a 5’cap and a 3’polyadenylated tail. Reading in the 5’to 3’direction, the PEDV genome contains genes for pol1 (P1) protein, spike (S) protein, an open reading frame (ORF3), envelope (E) protein, membrane (M) protein, and nucleocapsid (N) protein. Especially, ORF3 protein has been suggested as an important determinant for PEDV biological properties. In this study, ORF3 gene of attenuated DR13 strain was amplified by RT-PCR. Then, the ORF3 gene was cloned into pET-21a vector. The pET recombinants harboring ORF3 gene were transformed into E. coli BL21(DE3) host cells by electroporation. Protein expression was induced by isopropyl-beta-Dthiogalactoside and examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE showed a specific band of about 23.5 kDa in the whole cell lysate. The purified recombinant protein was evaluated for its antigenicity and reactivity through western blotting. The results indicated the recombinant ORF3 protein should be candidate as a feasible recombinant diagnostic reagent.