重组胸腺素α1的表达、纯化和生物学活性  

Expression,Purification and Bioactivity of Recombinant Thymosin α1

作　　者：苗红[1] 郭葆玉[1] 张冉[1] 张莉[1] 

机构地区：[1]第二军医大学药学院生化药学教研室,上海200433

出　　处：《中国生物化学与分子生物学报》2003年第5期636-639,共4页Chinese Journal of Biochemistry and Molecular Biology

摘　　要：为获得重组人胸腺素α1(recombinantthymosinα1,Tα1) ,采用融合表达方式表达Tα1基因 ,重组融合表达载体Tα1 pGEX 4XT 1转化大肠杆菌DE3(lys)构建工程菌 .对工程菌进行补料分批培养并诱导表达 ,得到目的蛋白的可溶性表达 .亲和层析纯化融合蛋白GST Tα1,经凝血酶裂解融合蛋白 ,亲和层析除去GST ,SourceQ离子交换 ,得到Tα1单体 ,得率为 30mg L发酵液 .生物学活性分析显示 ,重组Tα1能显著促进小鼠脾细胞增殖 (P <0 0 1) 。In order to produce recombinant thymosin α1(T α1), the recombinant plasmid T α1/pGEX 4T 1 was transformed into E.coli DE3(lys) and high level expression of soluble target protein was obtained by fed batch culture. The fusion protein was purified by affinity chromatography. T α1 was released after thrombin cleavage of the fusion protein. GST protein was removed by affinity chromatography, then T α1 was purified by ion exchange chromatography. The recovery rate of the final product was about 30 mg/L fermentation volume. Biological activity assay showed that purified T α1 could stimulate the proliferation of mice spleen cells with similar bioactivity to synthetic T α1.

关 键 词：重组胸腺素α1 表达 纯化 生物学活性 融合表达 

分 类 号：Q786[生物学—分子生物学] Q51