NDRG2截短体真核表达载体的构建

摘要

目的：构建稳定表达N-MYC下游调节基因2（NDRG2）截短体的真核表达载体。方法分别用PCR方法扩增NDRG2的截短体NDRG21～178aa和NDRG21～257aa ，以BamHⅠ和EcoRⅠ酶切后，以SolutionⅠ连接酶反应体系将NDRG2截短体片段装载入pCDNA4．0表达载体中，并通过蛋白免疫印迹（Western blotting）验证载体的表达。结果 NDRG2截短体在pCDNA4．0表达载体中正确表达。结论成功构建了NDRG2截短体的真核表达载体。%Objective To construct pCNDA4.0-NDRG2 vectors that correctly express the truncated N-MYC down-stream-regulated gene 2 (NDRG2).Methods The truncated NDRG21-178aa and NDRG21-257aa of NDRG2 was amplified by PCR, and were digested by BamHⅠand EcoRⅠ, then were cloned into the expression vector pCNDA 4.0 by SolutionⅠlig-ase reaction system.The protein expression of vectors was detected by Western blotting .Results The truncated NDRG2 was correctly expressed in the pCDNA 4.0 expression vector .Conclusion The eukaryotic expression vector of truncated NDRG2 is successfully constructed .