口蹄疫病毒VP1蛋白可溶性表达标签筛选

Screening of Fusion Tags that Enhanced Expression and Solubility of Type A Foot-and-mouth Disease Virus VP1 Protein

为了提高口蹄疫病毒VP1蛋白的可溶性表达,根据A型口蹄疫病毒核酸序列设计并合成了口蹄疫病毒VP1片段,将其克隆到p ET21b(+)载体上,设计1对通用引物扩增10个融合标签Grifin、GST、Nus A、SUMO、Thioredoxin、Protein G、γ-crystallin、Ars C、Ppi B、MBP,并分别将扩增的融合标签与VP1连接构建重组质粒,将重组质粒转化大肠杆菌BL21(DE3),0.5 mmol/L IPTG诱导其表达。SDS-PAGE结果表明,MBP、Grifin和GST能够促进口蹄疫病毒VP1蛋白的可溶性表达,其中MBP与VP1融合表达蛋白可溶部分占60%;Western blot鉴定分析结果显示,小鼠抗His标签单克隆抗体能识别表达的重组蛋白。表明成功表达了具有免疫学活性的融合蛋白MBP-VP1。

In order to improve the soluble expression of FMDV VP1 protein,FMDV VP1 gene fragment was designed and synthesized according to the nucleic acid sequence of type A FMDV,and was cloned into the vector p ET21b（ ＋）. A pair of universal primers and ten fusion tags（ Grifin,GST,Nus A,SUMO,Thioredoxin,Protein G,γ-crystallin,Ars C,Ppi B,MBP） were recombinanted with VP1 respectively,and then they were transformed into E. coli BL21（ DE3） and induced by 0. 5 mmol / L IPTG,respectively. The results of SDS-PAGE showed that MBP,Griffin and GST increased the solubility of the FMDV VP1 protein,which worked best with MBP,and the proportion of soluble cellular lysate was 60%. Western blot analysis showed that the recombinant protein could be recognized by mouse anti-his tag monoclonal antibody. It indicated that the fusion protein MBP-VP1 with immunological activity was successfully expressed.