%0 Journal Article
%T 一种新型双荧光素酶报告基因表达载体的构建<br>Construction of a novel dual luciferase reporter gene vector
%A 冯
%A 龙
%A 马云云
%A 曹
%A 珊
%A 李晓娟
%A 王媛媛
%A 陈肖楠
%A 孙倩倩
%A 吴建博
%A 赵国强
%J 郑州大学学报（医学版）
%D 2018
%R 10.13705/j.issn.1671-6825.2017.50.012
%X 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pGL4.10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pGL4.10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。<br>Aim: To construct a novel dual luciferase reporter gene vector.Methods: The rennet luciferase gene hRLUC was designed and amplified,then was inserted into pGL4.10 to obtain pGL4.10-hRLUC. pGL4.10 and pGL4.10-hRLUC were transfected into HEK293 cells using lipofectamine3000, and then the luciferase activity was detected. The cells without transfection were the control.Results: The hRLUC expression unit(2 350 bp)was amplified by PCR. The double luciferase reporter gene expression vector pGL4.10-hRLUC was identified by enzyme digestion and sequencing. The luciferase activity of cells transfected with pGL4.10-hRLUC was lower than those transfected with pGL4.10(220.311±2.082), higher than those without transfection(P<0.05).Conclusion: The novel double luciferase(hRLUC and Firefly)reporter gene vector is successfully constructed
%K 荧光素酶报告基因
%K 表达载体
%K hRLUC&lt
%K br&gt
%K luciferase reporter gene
%K vector
%K hRLUC
%U http://jms.zzu.edu.cn/oa/darticle.aspx?type=view&id=201801010