摘要
目的:克隆人Puma基因的启动子,插入荧光素酶报告基因载体中,并在细胞内检测其活性。方法:采用PCR技术,从人HepG2细胞中扩增出Puma启动子,插入荧光素酶报告基因载体pGL3-basic中,测序所扩增的DNA序列,并将其转染入H1299细胞中检测其活性。结果:测序结果表明,扩增的Puma启动子序列正确;双报告基因实验检测荧光素酶活力表明,构建的报告基因具有启动子活性。结论:Puma启动子的克隆及人Puma启动子报告基因的成功构建,为p53家族凋亡通路的功能研究提供了必要的实验材料。
Objective ：To construct the human Puma promotor lueiferase report gene vector and detect its activity in cells. Methods：The Puma promoter from HepG2 cell DNA was amplified by PCR,and was inserted into the lueiferase report gene pGL3 - basic vector. The amplified DNA sequence was confirmed hy sequencing and then the constructed vector was transfected into the H1299 cell to detect its activity by Premaga Dual - luciferase report gene detection system. Results ： The sequencing results indicated that the amplified sequence was correct, and the luciferase activity demonstrated that the constructed vector had the promotor activity. Conclusion：The human Puma promotor luciferase reporter gene vector has been constructed successfully, and it will become essential material for further study of the function of p53 family apoptosis pathway.
出处
《西南国防医药》
CAS
2009年第1期42-44,共3页
Medical Journal of National Defending Forces in Southwest China
