含p27启动子的荧光素酶报告基因的构建和活性测定  

Construction of a Luciferase Reporter Gene Vector Directed by p27 Promoter and Characterization of the Promoter Activity

作　　者：姜艳超[1] 丁丽华[1] 胡建民[2] 杨群辉[2] 曾敏[3] 杨智洪[1] 江泽飞[3] 李熔[1] 叶棋浓[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]沈阳农业大学,辽宁沈阳110161 [3]军事医学科学院附属医院,北京100071

出　　处：《生物技术通讯》2007年第2期186-188,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(30530320;30428012;30370738)

摘　　要：目的:克隆p27基因的启动子,插入荧光素酶报告基因载体中,并检测其活性。方法:采用PCR技术从人乳腺癌细胞系MCF-7基因组中扩增出p27启动子,插入荧光素酶报告基因载体pGL3-basic中,确定所扩增的DNA序列,在293T细胞中检测其活性。结果:测序结果显示扩增的p27启动子序列正确,活性实验表明构建的报告基因具有启动子活性,雌激素受体(ER)α能以剂量依赖的方式升高p27报告基因的转录。结论:克隆了p27启动子,为ERα共调节子的功能研究提供了重要基础。Objective： To clone p27 promoter and insert it into a luciferase reporter gene vector, and to characterize the promoter activity. Methods： The promoter of p27 was amplified from human breast cancer cell line MCF-7 genomic DNA by PCR and cloned into pGL3-basic, The resulting plasmid was determined by DNA sequencing. The promoter activity was analyzed in 293T cell line. Results： The result of DNA sequencing showed that the sequence of the cloned promoter was correct. The activity of the p27 luciferase reporter gene was confirmed, and this activity was increased by estrogen receptor（ER） α in a dose-dependent manner. Conclusion： The p27 promoter was cloned successfully, and this will be an important basis for the study of the functions of ERα coregulators.

关 键 词：p27启动子 荧光素酶报告基因 转录活性 

分 类 号：Q78[生物学—分子生物学]