Prokaryotic expression and immunogenicity analysis of S gene of Porcine epidemic diarrhea virus CH/ZJ strain

In order to obtain Porcine epidemic diarrhea virus S gene that has immune activity.The specific primers were designed by sequence of Porcine epidemic diarrhea virus(PEDV)CV777 strain S gene reported in GenBank excised signal peptide.The purpose gene of porcine epidemic diarrhea virus CH/ZJ strain was amplified by RT-PCR and then cloned into prokaryotic expression vector pET28a(+).The plasmid was transformed into E.coli Top10,and sequence was compared with PEDV CV777 strain.The gene comprised 966 bp encoding a polypeptide of 322 amino acids residues and the homology shared 96.0% nucleotide sequence with CV777,but deletion 3 bp at 398 bp position which encoding a isoleucine.The recombinant plasmid was transformed into E.coli BL21-DL3(Rosetta)and induced with IPTG.The expression product accounted for 80% of the total bacterial proteins by thin layer scanning.The recombinant protein was purified and then detected by SDS-PAGE and western-blot.The recombinant protein could react with the anti-PEDV hyperimmune serum.