羊痘病毒P32基因的克隆与原核表达

Cloning of the P32 Gene of Capripoxvirus and Construction of Its Prokaryotic Expression Vector

根据发表的羊痘病毒P32基因,设计一对特异性引物,PCR扩增P32全长基因,将其克隆入pMD-18-T载体,获得重组质粒pMD-P32。再将P32基因亚克隆入原核表达栽体pGEX-4T-1,获得重组表达质粒pGEX-P32,转化大肠埃希菌BL21后用IPTG诱导,表达产物经SDS-PAGE分析。结果表明,P32全长基因在大肠埃希菌中以融合形式成功表达,分子质量为58ku左右,与预期大小相符。

A pair of PCR primes were designed according to the published sequence of capripoxvirus major antigen p32. The P32 gene was multiplied by PCR and cloned into the pMD-18-T vector,we got the recombinant bacmid pMD-p32. Then the P32 gene was cloned into the E. coli expression vector pGEX-4T-1 and transformed into BL21 cells. After sequencing the gene,we got the recombinant plasmid pGEX-p32. After induction by IPTG in E. coli BL21, the 58 ku recombinant protein was obtained by SDS-PAGE.