摘要
根据GenBank上发表的羊痘病毒(CPV)P32基因序列,设计并合成一对引物,PCR扩增出837bp基因片段并克隆到pMD18-T载体中,酶切鉴定并测序。将P32基因定向克隆到pET32a表达载体中,酶切及测序鉴定正确后,转化BL21表达菌,经IPTG诱导得到了以包涵体形式表达的重组蛋白。采用镍离子亲和树脂在变性条件下对蛋白进行纯化,Western blot证明该重组蛋白具有良好的反应原性。
According to the published capripox virus （CPV） P3Z gene sequences in cenBanK,a pair ox prnii- ers were designed. 837 bp long 1932 gene fragment was amplified by PCR,cloned into pMD18-T vector,and identified by enzyme digestion and DNA sequencing. P32 gene was cloning into pET32a expression vector, expressed in BL21 bacteria and induced by IPTG. The recombinant protein was expressed in the form of in- clusion body. The recombinant protein was purified by nickel ion affinity chromatography in denaturing conditions. Western blot results showed that the recombinant protein had good immunogeriicity.
出处
《动物医学进展》
CSCD
北大核心
2013年第8期76-79,共4页
Progress In Veterinary Medicine
关键词
羊痘病毒
P32基因
表达
鉴定
Capripox virus
P32 gene
expression
identification
