人Cuedc2启动子区的克隆与鉴定  

Identification and Analysis of Human Cuedc2 Promoter

作　　者：刘淑娟[1] 王涛[2] 刘朵朵[1] 乔谷媛[1] 韩军涛[3] 

机构地区：[1]第四军医大学西京医院妇产科,陕西西安710032 [2]第四军医大学生物化学与分子生物学教研室,陕西西安710032 [3]第四军医大学西京医院烧伤科,陕西西安710032

出　　处：《现代生物医学进展》2014年第20期3828-3830,共3页Progress in Modern Biomedicine

基　　金：陕西省科技攻关基金项目(2011K12-10)

摘　　要：目的:克隆并鉴定和分析人Cuedc2启动子,为进一步研究其转录调控机制和功能提供实验基础。方法:对Cuedc2基因翻译起始位点上游约2000bp的序列进行在线生物信息学分析,使用PCR技术扩增该序列并测序,将扩增获得的该片段定向克隆入PGL-3basic载体中,构建荧光素酶报告基因质粒Cuedc2-luc。荧光素酶分析检测启动子的活性。结果:本实验成功构建了含有Cuedc2基因启动子序列的荧光报告系统,经体外验证该报告基因重组载体具有转录活性。结论:本实验所构建的Cuedc2基因启动子报告基因载体,为进一步研究Cuedc2基因的转录调控及其功能奠定了基础。Objective： To clone, identify and analyze human Cuedc2 promoter region and provide experimental basis for the research on the regulatory mechanism and function of Cuedc2. Methods： Appoximately 2000 bp up from the ATG of gene Cuedc2 was bioinfomatics analyzed online. The fragment was generated by polymerase chain reaction and sequenced for correctness. The Cuedc2-1uc promoter reporter was constructed by directly cloned this fragment into PGL-3basic vector and the promoter activity was measured by luciferase assay. Results： The fragment of 1982bp in upstream of the ATG, was amplified successfully and cloned directly into PGL-3basic vector to make Cuedc2 promoter reporter. Luciferase reporter assay showed that the fragment of 1956bp in length indeed had strong promoter activity. Conclusion： The luciferase reporter system of Cuedc2 gene promoter was constructed successfully and could be used for further study of the function and transcriptional regulation of Cuedc2.

关 键 词：CUEDC2 荧光素酶报告系统 转录调控 

分 类 号：R737.31[医药卫生—肿瘤] Q78[医药卫生—临床医学]