Cloning,sequencing and expression of ORF2 gene of porcine circovirus type 2 isolated from Jilin province of JL01 isolate

According to the nucleotide sequences of porcine circovirus type 2(PCV2) published in GenBank,a pair of primers specific to the ORF2 gene of PCV2 was designed and synthesized.ORF2 gene was amplified by polymerase chain reaction(PCR) from PCV2 Jilin strain(JL01) which was isolated from PK-15 cell line and had been characterized.The PCR product was cloned into the pMD 18-T vector and then sequenced.The recombinant plasmid pMD-ORF2 digested by Sall and XhoⅠ was subcloned into prokaryotic expression vector pET-32a.The resultant pET-ORF2 was transformed into E.coli BL21 cells and induced by IPTG.The result of SDS-PAGE and Western-blot indicated that the ORF2 gene had been expressed and could be recognized by the antiserum against PCV2.