猪细小病毒SH-1株VP2和NS1基因的克隆与原核表达

Molecular Cloning and Prokaryotic Expression of VP2 and NS1 Gene of Porcine Parvovirus SH Strain

通过PCR技术,从感染了猪细小病毒的ST细胞中扩增出猪细小病毒的VP2基因和NS1基因,并将其亚克隆至原核表达载体中,分别构建了重组表达载体pET32c/VP2和pET30a/NS1,经IPTG诱导,使pET32c/VP2和pET30a/NS1在受体菌BL21(DE3)中得到高效表达。经Western-blot检测具有生物学活性,为猪细小病毒野毒与疫苗毒的鉴别诊断奠定基础。

In this study , the open reading frame of PPV VP2 and NS1 were amplified by PCR method from the PPV infected ST cells. Then the products were cloned in prokaryotic vector and pET32c/VP2 and pET30a/NS1 expressed were construed respectively .The plasmids were introduced into E. coli BL21. After induction by IPTG , the high expression were found in products of pET32c/VP2 and pET30a/NS1 . Biological activity of the recombinant protein were detected by Western-blot , which can be applied in differential diagnosis of PPV infections.