冠状病毒HcoV-229E S1蛋白的原核表达及鉴定

目的克隆表达冠状病毒HcoV-229E S1基因片段,表达S1蛋白。方法合成冠状病毒HcoV-229ES1蛋白特异性基因片段并克隆入pET21a原核表达载体,转化BL21（DE3）菌,经IPTG高效诱导表达得到重组蛋白,用金属螯合亲和层析纯化,并通过Western blot对表达的重组蛋白进行鉴定。结果获得了主要以包涵体形式存在的目的蛋白,Western blot鉴定其为S1基因片段蛋白。结论成功构建了HcoV-229E S1蛋白的表达载体,并在BL21（DE3）中得到了高效表达,为下一步表达蛋白免疫原性及疫苗抗病毒保护性测定打下了基础。

Expression and identification of HcoV-229E coronavirus S1 protein in prokaryotic cell

Objective To clone S1 gene fragment of HeoV-229E eoronavims and express S1 protein. Methods S1 gene fragment was synthesized and cloned to pET21a vector. After sequencing,the S1 gene fragment was digested with Nde I and Xho I and inserted into prokaryotie expression vector pET-21a. The recombinant pET21a/HeoV-229E S1 was transferred into an expression strain E. coli BL21 （ DE3 ） and induced by IPTG express fusion protein. The protein was identified by Western blot and purified with a kit. Results It was found that the fusion protein of 14.4 kD was expressed mainly as inclusion body, The fragement of S1 protein was confirmed by Western blot. Conclusion It concludes prokaryotie expression vector pET21 a/HcoV-229E S1 is successfully constructed and S1 protein is expressed in E. coil BL21 （ DE3 ）. Our work may provide foundation for further research of protein immunogenieity and the detection of anti-vires vaccines.