重组人胸腺素β4二串体蛋白的原核表达、纯化及生物活性  

Prokaryotic Expression, Purification and Biological Activity of Re combinant Human Thymosin Beta 4

作　　者：徐天娇[1,2] 张昭[1,2] 舒震[1,2] 徐玉金[1,2] 赵薇[1,2] 李萌[1,2] 张伟 张英起[1,2] 

机构地区：[1]肿瘤生物学国家重点实验室 [2]第四军医大学药学系生物制药学教研室,陕西西安710032

出　　处：《生物技术通讯》2012年第3期329-332,共4页Letters in Biotechnology

基　　金：国家自然科学基金(31000406)

摘　　要：目的:获得具有生物学活性的重组人胸腺素β4二串体蛋白(Tβ4②)。方法:人工合成人Tβ4全长基因,构建Tβ4②基因,并将该基因克隆入载体pET-22b(+)中,转化宿主细胞大肠杆菌BL21(DE3),IPTG诱导表达后,经疏水作用层析纯化重组Tβ4②蛋白,采用Western印迹鉴定表达产物,并对其进行生物活性检测。结果:表达和纯化了Tβ4②蛋白,重组人Tβ4②在体外可促进人脐静脉内皮细胞的增殖和迁移。结论:重组人Tβ4②具有与化学合成Tβ4相同的功能,并有更高的生物学活性。Objective： To clone, express and purify recombinant human thymosin β4 tandem repeat（Tβ4②） in E.coli. Methods： A 135 bp tandem repeat of two Tβ4 gene was synthesized and cloned into pET-22b（＋） vector. The expression plasmid was then transformed into E.coli BL21 （DE3）. The expression of the recombinant Tβ4② was induced with IPTG. The expressed recombinant protein was purified by salting-out and hydrophobic chromatography and identified by SDS-PAGE and Western blot. The biological activity of Tβ4② was identified by MTT assay and in vitro migration assays. Results： A purified recombinant Tβ4② was obtained. The purified Tβ4② stimulated the proliferation and migration of HUVEC in vitro. Conclusion： The recombinant Tβ4②is successfully cloned, expressed and purified. The purified Tβ4② protein has the same function as synthesized Tβ4 but higher bioactivity.

关 键 词：胸腺素Β4 二串体 表达 纯化 活性检测 

分 类 号：Q78[生物学—分子生物学]