胸腺素α1二聚体蛋白的克隆、表达、纯化及其生物学活性检测
Cloning, expression, purification and biologic characteristics of Tα1 ② protein
李维娜 1吴守振 1张露 1韩苇 1颜真 2张英起1
作者信息
- 1. 第四军医大学药学系,生物制药教研室,肿瘤生物学国家重点实验室,陕西,西安,710033
- 2. 第四军医大学药学系,药物基因组学教研室,陕西,西安,710033
- 折叠
摘要
目的:获得具有生物学活性的重组人胸腺素α1二聚体蛋白.方法:人工合成胸腺素α1二聚体基因,并将该基因克隆入原核表达载体pET-22b(+)中.转化宿主细胞大肠杆菌BL21(DE3),经IPTG诱导表达重组Tα1②蛋白.对表达产物进行SDS-PAGE电泳和Western Blot检测分析以及生物学活性检测.结果:Tα1②Mr约为6.3×103,与理论值一致.成功纯化了Tα1②蛋白,该蛋白具有与T 1抗体特异性的结合能力并能刺激小鼠T淋巴细胞增殖.结论:成功地克隆、表达和纯化Tα1②蛋白;重组Tα1②具有与化学合成Tα1相同的功能并有更高的生物学活性.
Abstract
AIM: To clone, express and purify recombinant Thymosin alpha 1 ② protein (Tα1②) in E. coli. METHODS: A 189 bp recombinant gene of 2 × Tα1 tandem repeats was synthesized and cloned into pET-22b( + ) vector. The expression plasmid was then transformed into E. coll. BL21 ( DE3 ). The expression of the recombinant protein Tα1② was induced with IPTG and detected by SDS-PACE and Western blot. The biological activity of Tα1② was identified by MTT assay. RESULTS: A novel protein with expected molecular mass was expressed. The expressed product could be recognized by a monoclonal antibody against the human Tα1. The purified Tα1② stimulated the proliferation of mice splenic lymphocytes. CONCLUSION: The recombinant Tα1② is anccessfully cloned, expressed and purified. The purified Tα1 ②protein has the same function as synthesized Tα1 but higher bioactivity.
关键词
胸腺素α1/串联体/蛋白表达/纯化/活性检测Key words
thymsain alpha 1/concatemer/protein expression/purification/activity detection引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据