胸腺素α1串联体的表达、纯化及生物学活性检测

Expression of Thymosin alpha 1 Concatemer and Purification and Bioactivity of Expressed Product

目的 表达胸腺素α1 (Thymosinalpha1,Tα1 )三串体 ,并进行纯化及生物学活性鉴定。方法 使用大肠杆菌偏爱密码子 ,人工合成Tα1 三串体 (Tα1 ③ )基因 ,在表达载体pET -2 2b(+)中克隆 ,转化大肠杆菌BL2 1(DE3) ,IPTG诱导表达 ,经DEAE SepharoseFF阴离子柱纯化Tα1 ③蛋白 ,采用Westernblot鉴定表达产物 ,MTT法检测其生物学活性。结果 获得了纯化的Tα1 ③蛋白 ,相对分子质量约为 10 0 0 0 ,并具有Tα1 ③的生物学活性。结论 利用基因串联表达小分子多肽是一种可行的方法。

Objective To expresse thymosin alpha 1(Tα 1 ) c oncatemer,purify the expressed product and identify its bioactivity.Meth ods Synthesize Tα 1 concatemer using E.coli preferred codon,clone into expression vector pET-22b(+),transform to E.coli BL21 (DE3) and expre ssed under induction of IPTG.Purify the expressed product by DEAE-Sepharose FF anion exchange chromatography,identify by Western blot and determine its bioacti vity by MTT method. Results A purified Tα 1 protein with bioactivity and a relative m olecular weight of about 10 000 was obtained. Conclusion It is feasible to express the polypeptides with low relative molecular weights by gene cascade. [