Construction, Stable Expression of Survivin shRNA Vector

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摘要 Objective:ToconstructsurvivinshRNAexpressionvectorcartingenhancedgreenfluorescentproteingene,transfectitintoGBC-SDHcellsviaelectroporation,andgetGBC-SDcellswhicharestableexpressingsurvivinshRNA.Methods:ThesiRNAsequencetargetingsurvivinmRNAwassynthesizedandclonedintopEGFP-H1.TheconstructedplasmidandpEGFP-H1weretransfectedintoGBC-SDcellsrespectivelyvialiposome,andthetransfectingeffectwasdetectedwithFlowCytometry.ThenthetransfectedcellswereselectedwithG418.Results:Therecombinantplasmidwassuccessfullyconstructed,namedpEGFP-survivin.ThegenetransfectionefficienciesinpEGFP-H1-transfectedgroupandpEGFP-survivin-transfectedgroupwerethe80.29%±2.71%and83.85%±2.34%(P>0.05),whichwassuccessfultogetthecellsthatarestableexpressingshRNA,namedGBC-SD/EGFPandGBC-SD/survivin.Conclusion:SurvivinshRNAexpressionvectorwasconstructedsuccessfullyandgotGBC-SDcellswhicharestableexpressionshRNA.
机构地区 不详
出版日期 2011年01月11日(中国期刊网平台首次上网日期,不代表论文的发表时间)
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