细胞周期蛋白A2基因重组腺病毒载体的构建
China Health Care & Nutrition(2017)
Abstract
目的:构建并鉴定细胞周期蛋白A2基因重组腺病毒AD-Cyclin A2.方法:选用聚合酶链反应(PCR)法对基因CyclinA2进行扩增,并应用AdMaxTM包装系统重组载体质粒AD-eGFP-CyclinA2,经测序鉴定重组后转染HEK293细胞,进行腺病毒的包装、扩增与纯化,最后测定腺病毒滴度.结果:经酶切、序列分析鉴定后,重组腺病毒载体AD-eGFP-Cyclin A2成功构建,经转染的HEK293细胞出现了明显的细胞病变效应,且获得重组腺病毒的滴度为3.51×108IFU/mL.结论:成功获得重组腺病毒AD-Cyclin A2,也为今后临床研究基因治疗奠定实验基础.
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