摘要
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）,VP1 gene of SVA was cloned and sequenced.A phylogenetic tree was constructed based on VP1 gene, amino acid mutations were analyzed and the secondary structure of VP1 protein was predicted. The VP1 gene of SVA was cloned into p ET-32a（＋） and expressed with IPTG induction. The recombinant VP1 protein was purified by Ni-NTA affinity chromatography and identified with Western-blot. The results show that the recombinant VP1 protein was48 ku in molecular weight, and mainly expressed as inclusion bodies.It could specifically react with anti-His monoclonal antibody. The results indicated that the VP1 protein was successfully expressed and purified.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第4期500-507,共8页
Chinese Veterinary Science
