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龙源期刊网 http:/www.qikan.com.cnPRDX5 基因启动子的荧光素酶报告质粒的 构建作者:胡乐林 来源:医学信息2014年第 16期摘要:目的构建含PRDX5基因启动子的荧光素酶报告质粒。方法通过生物信息学方 法,查询PRDX5基因启动子核酸序列;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;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 PromoterHU 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. Methods search the nucleic acid sequence of the promoter PRDX5 by bioinformatics analysis. Methods 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. pGL3-PRDX5-promoter-Luc plasmid was verified by doubleenzyme cleavage and sequencing method.ResultspGL3-PRDX5-promoter-Luc plasmid was identified to be correct by double-enzyme cleavage and sequencing method.Conclusion A pGL3-PRDX5- promoter-Luc plasmid was successfully constructed.Key words: PRDX5;Promoter;Luciferase reporter plasmid;Reactive oxygen speciesPRDX5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 shiota2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粒体的PRDX5能保护线粒体DNA免受过氧化氢的损伤。并 且PRDX5能够抑制p53诱导的活性氧簇的产生和细胞凋亡3。1材料与方法
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