SARS病毒S蛋白S1和S2片段的合成及在大肠杆菌中的表达

摘要

[Objective] To obtain pure recombinant SI and S2 of SARS S protein. [ Method] Using asymmetric PCR and ligation with endonu-clease,Sl 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-Sl and pET28a-S2, respectively. These recombinant vectors were transformed into E. Coli BL21 ,and expression of SI and S2 fragments were induced by IPTG. The conditions of expression and purification were optimized. [ Result] The SI and S2 fragments were amplified and successfully expressed in E. Coli. [ Conclusion] This research provides detection antigens for follow-up development of SARS vaccine.%[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.