含人SOD_2基因启动子的荧光素酶报告基因质粒的构建和鉴定  

The construction and identification of a luciferase reporter gene vector directed by human SOD_2 promoter

作　　者：李新娜[1] 史艳芬[2] 仲苓芝[1] 李文雪[1] 李荣贵[1] 

机构地区：[1]吉林大学白求恩医学院病理生物学教育部重点实验室,吉林长春130021 [2]北京中日友好医院,北京100029

出　　处：《吉林医学》2012年第34期7399-7400,共2页Jilin Medical Journal

摘　　要：目的:从人基因组中克隆SOD2基因的启动子并插入到荧光素酶报告基因载体中并进行基因测序鉴定。方法:以人基因组DNA为模板,用PCR的方法扩增出人SOD2基因启动子片段后,将其插入到pGL3-basic载体的荧光素酶报告基因上游,将构建的重组质粒通过双酶切片段的凝胶电泳分析和DNA测序以确定插入位置和序列的正确性。结果:双酶切反应和测序结果显示SOD2基因启动子插入的位置和序列是正确的。结论:成功克隆了SOD2基因启动子,为后续SOD2转录调控机制的研究提供重要的实验基础。Objective To clone the promoter of human superoxide dismutase 2 gene and insert it into a luciferase reporter gene vector, and then to identify and check the gene sequence. Method The promoter of the human SOD2 gene was amplified from human genomic DNA by PCR,which was cloned into pGL3 -basic vector. Recombinant plasmid was cloned screening and amplified. The plasmid was determind- ed by double digestion reaction and DNA sequencing. Results The double digestion reaction and DNA sequencing showed that the sequence of cloned promoter was right and its gene locus was correct. Conclusion The cloning of the promoter of human SOD2 gene has been done successfully ,and it will be a important basis for the following study of SP1 transcription factors interacting about vascular injury by Arsenic.

关 键 词：人超氧化物歧化酶2(SOD2) 荧光素酶 启动子 报告基因 

分 类 号：R599[医药卫生—内科学]