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为表达并纯化塞内加谷病毒(Senecavirus A,SVA)的VP1蛋白,克隆SVA的VP1基因,测定其序列并构建遗传进化树,分析VP1蛋白氨基酸突变位点并预测其二级结构。然后将VP1基因克隆至载体pET-32a(+)中,IPTG诱导表达VP1重组蛋白,用Ni-NTA亲和层析柱纯化并用W estern-blot进行验证。结果显示,表达的VP1重组蛋白的大小为48 ku,主要以包涵体形式存在,能与抗His标签的单克隆抗体发生特异性免疫反应。结果表明成功表达并纯化出VP1重组蛋白。
To express and purify the VP1 protein of Senecavirus A (SVA), the VP1 gene of SVA was cloned. The sequence of the VP1 gene was determined and the phylogenetic tree was constructed. The amino acid mutation site of VP1 protein was analyzed and its secondary structure was predicted. The VP1 gene was then cloned into vector pET-32a (+). VPT recombinant protein was induced by IPTG, purified by Ni-NTA affinity chromatography and verified by Western-blot. The results showed that the recombinant VP1 protein was 48 ku in size and mainly existed in the form of inclusion body, which could specifically react with anti-His tag monoclonal antibody. The results showed that VP1 recombinant protein was successfully expressed and purified.