prdx5基因启动子的荧光素酶报告质粒的构建.doc

PRDX5基因启动子的荧光素酶报告质粒的构建
摘要:目的构建含PRDX5基因启动子的荧光素酶报告质粒。方法①通过生物信息学方法,查询PRDX5基因启动子核酸序列;②使用PCR扩增人PRDX5基因启动子片段,将启动子插入到pGL3-Basic载体中,构建含PRDX5基因启动子片段的荧光素酶报告质粒;③双酶切及测序确定PRDX5基因启动子的荧光素酶报告质粒扩增正确。结果 PCR扩增的PRDX5启动子片段插入到载体中,经测序证实正确。结论成功构建含PRDX5启动子的荧光素酶报告质粒。
关键词:PRDX5;启动子;荧光素酶报告基因
Construction of Luciferase Reporter Plasmid PRDX5 Gene Promoter
HU Le-lin
(School of Medicine,The Anhui University of Science and Technology,Huainan 232001,Anhui,China)
Abstract:ObjectiveTo construct a pGL3-PRDX5-promoter-Luc plasmid and to vertifying its luciferase activity. Methods①search the nucleic acid sequence of the promoter PRDX5 by bioinformatics analysis.②Methods of PCR was performed to amplify the promoter PRDX5. the promoter PRDX5 was inserted into pGL3-Basic cloning vector to construct luciferase reporter plasmid with the promoter PRDX5.③pGL3-PRDX5-promoter-Luc plasmid was verified by double-enzyme cleavage and sequencing -PRDX5-promoter-Luc plasmid was identified to be correct by double-enzy
me cleavage and sequencing A pGL3-PRDX5-promoter-Luc plasmid was essfully constructed.
Key words:PRDX5;Promoter;Luciferase reporter plasmid;Reactive oxygen species
PRDX5是Peroxiredoxins(PRDXs)过氧化物酶家族蛋白的一种,研究显示在它广泛表达于哺乳动物的各种细胞,定位于细胞质、过氧化物酶体和线粒体中,但在许多细胞中PRDX5主要定位于线粒体,它与组织过氧化物、过氧化亚硝基阴离子有高的亲和力[1]。Masaki shiota[2]等人发现在外源过氧化氢处理或组织缺氧情况下人类前列腺癌PC3细胞PRDX5表达增高。研究还显示人类定位于线