猪繁殖与呼吸综合征病毒M基因原核表达载体的构建与表达

Construction and expression of prokaryotic expression vector of M gene of porcine reproductive and respiratory syndrome virus

根据GenBank上已公布的猪繁殖与呼吸综合征病毒(PRRSV)核酸序列设计1对引物,用RT-PCR方法扩增出去除部分疏水区的PRRSV-M基因,并将其克隆到原核表达载体PET-30a上,得到重组质粒PET-30a-M。将重组质粒转化大肠杆菌transetta(DE3)感受态细胞,经IPTG诱导表达、SDS-PAGE电泳及Western blot分析,显示重组蛋白分子量约1 5.7 kD,且此蛋白能与His标签单克隆抗体发生特异性反应。

A pair of primers were designed according to the nucleotide sequence of porcine reproductive and respiratory syndrome virus（PRRSV）published in the Genbank, and the M gene of PRRSV was amplified by RT-PCR and then cloned into the prokaryotic expression vector PET-30a. The obtained recombinant plasmid PET-30a-M was transferred to E. coli transetta（DE3）competent cell and expressed via IPTG induction. The SDS-PAGE and Western blot analysis showed that the cloned PET-30a-M/E. coli produced about 15.7 kD of recombinant protein which could produce a specific reaction with His-Tag monoclonal antibody.