IFITM1基因真核表达质粒的构建及鉴定

Construction and identification of a eukaryotic expression plasmid encoding IFITM1 gene

目的:获得IFITM1基因片段并构建真核表达质粒。方法:采用RT-PCR技术扩增IFITM1,将扩增产物连接至pcDNA3.1载体,对重组质粒进行测序验证,结果:构建了真核表达质粒pcDNA3.1-IFITM1,通过酶切、测序等方法验证完全正确。结论:成功构建了IFITM1基因的真核表达质粒,为下一步探讨IFITM1基因在子宫颈癌HeLa细胞中的作用提供了实验基础。

Objective：To obtain IFITM1 gene fragment and to construct eukaryotic expression plasmid.Methods：IFITM1 gene was amplified by RT-PCR technique and PcDNA3.1 eukaryotic expression plasmid was isolated.IFITM1 gene and PcDNA3.1 eukaryotic expression plasmid were digested by BamH1/EcoR1 restriction endonuclease.IFITM1 gene and PcDNA3.1 was connected by T4 DNA ligase.Lastly the recombinant plasmids were transformed to Dh5αcompetent cells and these recombinant plasmids were identified by restriction endonuclease and sequencing.Results：Eukaryotic expression IFITM1-PcDNA3.1 recombinant plasmids were successfully constructed and the plasmids were completely corrected by restriction endonuclease and sequencing.Conclusion：Successfully construct IFITM1-PcDNA3.1 recombinant plasmids,it provides research materials for further studying the function of IFITM1 gene in HeLa cells.