Hes1基因启动子的克隆及活性研究

Cloning and activity analysis of human Hes1 gene promoter

目的:获取人Hes1基因的启动子并对其进行活性分析。方法:以人Hela细胞基因组DNA为模板,利用PCR技术扩增Hes1基因5'侧翼序列,将扩增的片段与pMD18-T载体连接,然后利用双酶切进行鉴定,并送测序;将测序正确的启动子插入pGL3-Basic,双酶切并测序鉴定。将测序正确的重组DNA质粒瞬时转染宫颈癌细胞,运用双荧光素酶报告基因检测系统检测其启动子的活性。结果:PCR扩增到的人Hes1基因启动子与基因库的序列完全一致,双荧光素酶报告基因检测系统显示其在宫颈癌细胞中具有启动子活性。结论:成功得到人Hes1基因启动子并且其具有活性。

Objective： To get the human hesl gene promoter and to analyze its aetivity. Methods： With Hela genomic DNA as a template, amplify Hesl 5 ＇flanking sequenees with PCR, and use double enzyme and sequence to identify it whieh the amplified fragment connected with pMD18-T vector. Insert the right one to pGL3-Basic; also identify it with double enzyme and sequence. To sequencing the right reeombinant DNA plasmid transient transfeetion of cervical cancer cells, using dual lueiferase reporter gene detection system to deteet the activity of the promoter. Results：Sequence of the human Hesl promoter,which amplify with PCR, is just the same as its in Gene Pool, also detect its activity with dual luciferase reporter gene detection system. Conclusions：Get the activated human Hesl gene successfully.