关键词: 人Era基因;基因表达;蛋白纯化

    摘 要:目的 在大肠杆菌中对人的era基因(简称hera)进行表达、纯化. 方法 PCR扩增hera基因，测序正确后克隆入大肠杆菌融合表达载体pRSET-C，重组质粒pRSETC-hera以大肠杆菌BL21(DE3)为宿主菌，用IPTG进行诱导表达.然后利用亲和层析的方法对表达蛋白进行纯化. 结果 克隆了hera基因，测序正确;利用pRSET-C载体在大肠杆菌中融合表达了全长hera-cDNA基因，表达量占细菌总蛋白的59.6%.融合蛋白在菌体内主要以包涵体形式存在，在变性条件下用Ni-NTA亲和色谱柱纯化此融合蛋白，其纯度达到了98.0%. 结论 利用基因重组技术在大肠杆菌中高表达了hera基因.并对融合蛋白进行了初步纯化.

    High expression of human Era protein and its pu┐rification

    WU Yuan-Ming，CHEN Su-Min，CHEN Nan-Chun，JI Zong-Lin，LIU Hui-Ping，ZHANG Jun-Jie Department of Biochemistry and Molecular Biology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China

    Keywords:human Era gene;gene expression;protein purifi-cation

    Abstract:AIM To express human Era protein in E.coli and purify it preliminarily.METHODS After being amplified by PCR and identified by sequencing the human era gene was in-serted into expression vector pRSET-C.The recombinant plasmid pRSETC-hera was transformed into BL21(DE3)and induced with IPTG.The expressed protein was purified by affinity chromotography.RESULTS The human era gene was amplified and the sequence proved to be correct.The hera-cDNA gene had been cloned into the vector and ex-pressed in E.coli ......