一种新型双荧光素酶报告基因表达载体的构建

Long FENG,Yunyun MA,Shan CAO,Xiaojuan LI,Yuanyuan WANG,Xiaonan CHEN,Qianqian SUN, Jianbo WU,Guoqiang ZHAO

Journal of Zhengzhou University(Medical Sciences)（2018）

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Abstract

目的:构建一种新型的双荧光素酶报告基因表达载体.方法:设计并扩增海肾荧光素酶基因hRLUC,将其插入到双酶切的含有萤火虫荧光素酶报告基因(Firefly)的载体pGI4.10中,得到pGL4.10-hRLUC;使用lipofectamine3000将pGL4.10和pGL4.10-hRLUC分别转染入HEK293细胞,检测细胞中荧光素酶活性,以未转染细胞作对照.结果:使用PCR方法获得hRLUC表达单元(2 350 bp);双荧光素酶报告基因表达载体pGL4.10-hRLUC经酶切和测序鉴定,插入正确.转染pGL4.10组荧光素酶活性相对值为(220.311 ±2.082);转染pGL4.10-hRLUC的细胞中荧光素酶活性低于pGI4.10转染细胞,高于未转染细胞(P＜0.05).结论:成功构建了双荧光素酶报告基因(hRLUC和Firefly)表达载体.

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Key words

luciferase reporter gene,vector,hRLUC

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