小鼠ERA蛋白在大肠杆菌中的表达及其纯化  

Expression and purification of mouse ERA protein in Escherichia coli

作　　者：杜孟刚[1] 陈苏民[1] 吴元明[1] 董珂[1] 陈南春[1] 郑玉[1] 

机构地区：[1]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2004年第8期684-687,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金资助项目 (39870 380 ;39670 0 0 6) ;全军医药卫生科研基金资助项目 (98M1 0 8)

摘　　要：目的 :对小鼠ERA蛋白 (mERA)进行表达、纯化 ,为真核生物ERA功能研究奠定基础 .方法 :构建MBP mER A融合表达载体 .在大肠杆菌中诱导表达融合的小鼠ERA蛋白 ,经直链淀粉亲和层析纯化 ,WesternBlotting鉴定所表达纯化的蛋白 .结果 :表达的MBP mERA融合蛋白占菌体总蛋白的 1 8% ;纯化后的融合蛋白纯度为 70 % ;WesternBlotting证实了mERA蛋白的表达 .结论 :利用基因重组技术 ,在大肠杆菌中高表达了mera基因 ,并对融合蛋白进行了初步纯化 .AIM: To express and purify mouse ERA protein for the further study of eukaryotic ERA function. METHODS: The fused protein expression vector pMSM mERA was constructed. The MBP mERA was expressed in E. coli after induction and the target protein was purified by means of amylose affinity chromatography and identified by Western blotting. RESULTS: The expressed MBP mERA protein accounted for 18% of the total bacterial protein and was identified by Western Blotting. The purity of the fused protein was 70 % after amylose affinity chromatography. CONCLUSION: The fused mera gene can be expressed with high efficiency in E.coli by gene recombination technique and the fused protein is purified preliminarily.

关 键 词：小鼠 ERA基因 基因表达 蛋白质类 分离和提纯 

分 类 号：Q786[生物学—分子生物学]