SARS病毒S蛋白S1和S2片段的合成及在大肠杆菌中的表达  

Synthesis,Expression and Purification of S1 and S2 Fragments of SARS S Protein in E.coli

作　　者：郑尚永[1] 潘卫庆[1] 

机构地区：[1]中国人民解放军第二军医大学病原生物教研室,上海200433

出　　处：《安徽农业科学》2011年第20期12208-12211,共4页Journal of Anhui Agricultural Sciences

摘　　要：[目的]通过拼接完成对SARS S蛋白的上下游序列S1和S2的合成,并在大肠杆菌中表达,获得具活性的S1和S2蛋白。[方法]利用不对称PCR方法以及采用设计酶切位点将2个片段连接的方法合成SARS香港株S蛋白上下游2个片段S1和S2;构建S1和S2这2个基因片段的原核表达载体,pET28a-S1和pET28a-S2,并转化BL21表达菌株,然后用IPTG诱导目的基因的表达并提取纯化目的蛋白。[结果]连接得到S1和S2这2个基因片段,并成功地在大肠杆菌中得到表达。[结论]该研究为后续SARS疫苗研究提供检测抗原奠定了基础。[Objective] To obtain pure recombinant S1 and S2 of SARS S protein.[Method] Using asymmetric PCR and ligation with endonuclease,S1 and S2 fragments of SARSV HK strain S gene were synthesized.Then,these two fragments were inserted into plasmid pET28a to obtain recombinant vectors pET28a-S1 and pET28a-S2,respectively.These recombinant vectors were transformed into E.coli BL21,and expression of S1 and S2 fragments were induced by IPTG.The conditions of expression and purification were optimized.[Result] The S1 and S2 fragments were amplified and successfully expressed in E.coli.[Conclusion] This research provides detection antigens for follow-up development of SARS vaccine.

关 键 词：SARS S蛋白 S1蛋白 S2蛋白 原核表达 纯化 

分 类 号：S856.3[农业科学—临床兽医学]