抑癌基因FHIT真核表达质粒的构建

Construction of eukaryotic expression vector of tumor suppressor gene FHIT

目的 构建抑癌基因FHIT真核表达质粒pHCMVsp1A FHIT。方法 用限制性内切酶EcoRI酶切抑癌基因FHIT的克隆载体pGEM FHIT获得FHIT基因片段 ,长度为 0 9kb ,将其插入真核表达载体pHCMVsp1ACMV启动子的下游 ,用HindIII酶切鉴定FHIT基因插入方向。结果 PCR扩增及EcoRI、HindIII酶切鉴定证实FHIT基因以正确方向插入表达质粒pHCMVsp1A中。结论 成功构建抑癌基因FHIT真核表达质粒pHCMVsp1A FHIT 。

Objective: To construct an eukaryotic expression vector of tumor suppressor gene FHIT. Methods: The full length cDNA fragment of FHIT was obtained by the digestion of cloning vector pGEM FHIT with EcoRI,then cloned into eukaryotic expression vector pHCMVsp1A under the CMV promoter.The recombinant plasmid DNA was identified by the digestion of HindIII and PCR. Results: The FHIT cDNA was cloned into the pHCMVsp1A in the right direction. Conclusion: The eukaryotic expression vector pHCMVsp1A FHIT has been constructed successfully and it may be used for further research on the tumor suppressor function of FHIT.