Annexin A2原核表达载体的构建及其重组蛋白的纯化

Construction of Annexin A2 expression vector and purification of Annexin A2 recombinant protein

目的构建Annexin A2原核表达载体并纯化其重组蛋白,用于进一步研究Annexin A2的生物学功能及其互作蛋白分析。方法经基因重组技术构建Annexin A2原核表达载体,经测序鉴定、转化至E.coli BL21菌株后,诱导表达Annexin A2重组蛋白,并获得纯化的重组蛋白。结果载体克隆序列及开放阅读框架正确,经IPTG诱导在62kDa处获得重组蛋白,纯化后得到大量GST-Annexin A2重组蛋白。结论成功构建了pGEX-Annexin A2原核表达载体,表达并纯化出GST-Annexin A2融合蛋白。

Objective To construct the prokaryotic expression vector of Annexin A2 and purify its recombinant protein for further functional study.Methods The prokaryotic Annexin A2 vector was constructed with recombinant techniques.After identificated by DNA sequencing and transformed into E.coli BL21,the Annexin A2 vector was induced by IPTG.Then the fusion protein was purified by GST-pull down.Results Both the sequences and the open reading frame of the vector were correct.The fusion protein showed at 62kDa.GST-Annexin A2 fusion protein was purified by pull-down.Conclusion The pGEX-Annexin A2 expression plasmid was successfully constructed and the recombinant GST-Annexin A2 fusion protein could be expressed and purified.