重组SARS冠状病毒S蛋白的原核表达研究

Expression and purification of recombinant SARS coronavirus spike protein

目的克隆表达SARS冠状病毒的主要结构蛋白(S蛋白)。方法合成SARS冠状病毒S蛋白特异性基因片断并克隆入pET32a原核表达载体,转化BL21菌,经IPTG诱导高效表达得到重组S蛋白,并通过W estern印迹对重组蛋白质进行鉴定。结果重组蛋白质经镍柱亲和层析得到了部分纯化,免疫动物后得到SARS病毒的多克隆抗体。结论经E lisa检测,表达的重组S蛋白基本具备检测病人血清中抗SARS病毒IgG和IgM的能力,可进一步用于S蛋白功能研究与SARS诊断试剂盒的研制。

Aim To clone and express the Spike protein of SARS-CoV. Methods In this work, clone a special gene fragment of S protein of SARS-CoV into vector pET32a, lnduce the expression efficiently by IPTG after the express vector transforms E. coli BL21. Identify the recombinant protein of gain by Western blot. Results The recombinant protein was run through a pillar of nickel to turn pure, to immunize rabbit to get the polyclonal antibody. Conclusion Examined by Elisa, the purified protein can examine the IgG and the IgM in SARS-patient＇s serum. It can be further used for the study on Sprotein＇s function research and the development of diagnostic method of SARS.