人胸腺素α1原核表达重组体的构建及其在大肠杆菌中的表达

Construction of Prokarocyte Expression Recombinant of Human Thymosin α1 and Its Expression in Escherichia coli

目的 :以基因工程方法获得人胸腺素 α1 (Tα1 )多肽产品。方法 :通过 PCR扩增人工合成模板的方法获得人 Tα1基因 ,然后将其克隆入原核细胞表达载体 p GEMEX1 ,用重组体转化大肠杆菌 ,并从 m RNA和蛋白质水平检测重组体在大肠杆菌中的表达情况。结果 :构建了人 Tα1克隆重组体 p UC1 8-Tα1 ,经序列分析证实对码、序列无误 ;成功构建了人 Tα1原核细胞表达重组体 p GE-MEX1 -Tα1 ;从 m RNA水平证实 p GEMEX1 -Tα1在大肠杆菌 JM1 0 9(DE3)中能够转录 ;转化大肠杆菌 JM1 0 9(DE3) ,经 IPTG诱导 ,能产生一 36 k D左右的融合蛋白。结论 :构建成功重组体 p GE-MEX1 -Tα1 。

Objective: To acquire human thymosin α1(Tα1) peptides by gene engeering techniques.Methods: Tα1 gene was acquired by molecular biological techniques and cloned into vector pGEMEX1 Tα1.An appropriate Escherichia coli (E.coli) strain was transformed by the recombinant pGEMEX1 Tα1 and its transcription and expression in E.coli we tested.Results: pUC18 Tα1,the cloning recombinant of human Tα1,was constructed and confirmed by sequencing,and pGEMEX1 Tα1,the prokarocyte expression recombinant of human Tα1,was successfully constructed.Then it was confirmed that pGEMEX1 Tα1 could be transcribed in JM109 (DE3).A 36 kD fusion protein was produced after JM109 (DE3) transformed with pGEMEX1 Tα1 and induced with IPTG.Conclusion: The recombinant pGEMEX1 Tα1 can be successfully constructed and expressed in E.Coli.