人正、反义转化生长因子beta1基因真核表达载体的构建

目的 :构建人转化生长因子beta 1(TGFβ1)正、反义基因真核表达载体。方法 :从人胎脑的cDNA文库中扩增出TGFβ1共 12 84bp长的DNA片段后 ,与T载体直接进行高效连接 ,并测序证实无突变。接着利用TGFβ1引物两端所设计的酶切位点将目的片段定向亚克隆到真核表达载体pcDNA3.1( )。构建的正反义表达重组体 (pcDNA3.1 TGFβ1和pcDNA3.1 antiTGFβ1)经限制性内切酶消化证实其中有目的片段完整插入。结果 :本实验成功构建了人正、反义TGFβ1基因真核表达载体。结论 :人正、反义TGFβ1基因真核表达载体的构建 ,为今后研究腹膜纤维化的防治奠定了实验基础。

Construction of sense and antisense expression vector for human transforming growth factor-beta 1

Objective To construct transforming growth factor beta 1 ( TGFβ1) sense and antisense eukaryotic expression vector. Methods The TGFβ1 cDNA fragments with 1 284 base pairs were obtained by PCR from human fetal brain cDNA. The efficient link was carried out directly between the TGFβ1 cDNA and the vector using PGEM T vector system,which was confirmed by DNA sequencing. Then the target fragment was subcloned into eukaryotic expression vector pcDNA3.1( ) in orientation.The forward and reverse inserting recombinants were confirmed by restriction endonuclease digestion ( EcoR Ⅰand Hind Ⅲ).Results Recombinant human TGFβ1 sense and antisense expression vectors were constructed successfully.Conclusion The human TGFβ1 sense and antisense expression vectors may be the experimental basis of peritoneal fibrosis study.