塞内加谷病毒VP1蛋白的表达纯化及鉴定  

Expression, purification and identification of VP1 protein of Senecavirus A

作　　者：白杨[1] 伍绮文 陈桂华[1] 周玲[1] 李怡昀 周雯馨 马若瑶 马静云[1] 

机构地区：[1]华南农业大学动物科学学院,广东广州510642

出　　处：《中国兽医科学》2017年第4期500-507,共8页Chinese Veterinary Science

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.

关 键 词：塞内加谷病毒 VP1蛋白 表达 纯化 

分 类 号：S852.659.6[农业科学—基础兽医学]