胸腺素α1的基因合成、表达及活性测定

Synthesis and Expression of Thymosin α1 Gene and Biological Activity of Expressed Product

目的 合成胸腺素α1(Tα1)基因并在原核细胞中表达。方法 用半酶半化学合成并克隆Tα1基因 ,将该基因插入表达质粒pGEX 4T 1并在大肠杆菌进行表达 ,对表达的融合蛋白GST Tα1利用亲和层析法纯化 ,再经Thrombin酶切后 ,获得重组Tα1进行生物学活性检测。结果 经测序证实合成的Tα1序列与设计的一致。构建的重组表达载体pGEX 4T 1 Tα1在大肠杆菌中得到高效表达。E玫瑰花结形成试验证明重组Tα1具有生物学活性。结论 合成的Tα1基因在大肠杆菌中表达出具有生物活性的蛋白。

Objective To synthesize and express thymosin α1 (Tα1) gene in prokaryocyte.Methods Tα1 gene was synthesized and clone by zymological and chemical methods and inserted into expression plasmid pGEX 4T 1,then transformed to E.coli .The expressed fusion protein GST Tα1 was purified by affinity chromatography,digested with thrombin,and detected for biological activity.Results The DNA sequence of the synthesized Tα1 gene was identical to that designed.The constructed recombinant plasmid pGEX 4T 1 Tα1 was highly expressed in E.coli .E rosette test showed biological activity of the recombinant Tα1.Conclusion The synthesized Tα1 gene was successfully expressed in E.coli ,and the expressed protein showed biological activity.