猪繁殖与呼吸综合征病毒nsp7蛋白的表达与纯化

摘要

To investigate the immunological activity of nsp7 recombinant protein,the PRRSV BJ-4 nsp7 gene amplified by RT-PCR was sub-cloned into the Escherichia coli expression vector pET-28a. The recombinant protein was expressed in E.coli BL21 (DE3) and purified by Ni-chelation. After purification,the recombinant protein was analyzed by SDS-PAGE and western blot. The im-munogenicity of nsp7 protein was evaluated by ELISA. The results showed that the nsp7 protein was expressed successfully and the recombinant protein had immunological activity which could be recognized by the porcine positive serum. This assay provided theoretical support for construc-ting indirect-ELISA of nsp7 recombinant protein.%为了解nsp7重组蛋白的免疫学活性,采用RT-PCR的方法从猪繁殖与呼吸道综合症病毒(PRRSV)BJ-4毒株中扩增得到nsp7基因,将基因连接到pET-28a栽体并转化至大肠杆菌BL21中进行诱导表达,利用Ni-NTA进行亲和纯化,通过SDS-PAGE和Western-blot对重组蛋白进行分析和鉴定,通过间接ELISA方法研究了重组蛋白的免疫学活性.结果表明,成功制备了nsp7重组蛋白,间接ELISA结果表明,重组蛋白具有良好的免疫活性,为建立nsp7特异性抗体的间接ELSIA检测方法及PRRS诊断试剂盒的研制奠定了基础.