山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2014年
23期
9-12
,共4页
李惠晨%马勇政%王泽宇%胡伟%张健
李惠晨%馬勇政%王澤宇%鬍偉%張健
리혜신%마용정%왕택우%호위%장건
N-MYC下游调节基因2%截短突变体%pCNDA4.0载体
N-MYC下遊調節基因2%截短突變體%pCNDA4.0載體
N-MYC하유조절기인2%절단돌변체%pCNDA4.0재체
N-MYC downstream-regulated gene 2%truncated mutants%pCNDA4.0 vector
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。
目的：構建穩定錶達N-MYC下遊調節基因2（NDRG2）截短體的真覈錶達載體。方法分彆用PCR方法擴增NDRG2的截短體NDRG21～178aa和NDRG21～257aa ，以BamHⅠ和EcoRⅠ酶切後，以SolutionⅠ連接酶反應體繫將NDRG2截短體片段裝載入pCDNA4．0錶達載體中，併通過蛋白免疫印跡（Western blotting）驗證載體的錶達。結果 NDRG2截短體在pCDNA4．0錶達載體中正確錶達。結論成功構建瞭NDRG2截短體的真覈錶達載體。
목적：구건은정표체N-MYC하유조절기인2（NDRG2）절단체적진핵표체재체。방법분별용PCR방법확증NDRG2적절단체NDRG21～178aa화NDRG21～257aa ，이BamHⅠ화EcoRⅠ매절후，이SolutionⅠ련접매반응체계장NDRG2절단체편단장재입pCDNA4．0표체재체중，병통과단백면역인적（Western blotting）험증재체적표체。결과 NDRG2절단체재pCDNA4．0표체재체중정학표체。결론성공구건료NDRG2절단체적진핵표체재체。
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 .
