山东医药2017，Vol.57Issue(4)：1-4,4.DOI:10.3969/j.issn.1002-266X.2017.04.006
人 Tudor-SN UTR 片段荧光素酶报告基因表达质粒的构建及活性检测
Construction of human Tudor-SN UTR luciferase reporter gene expression plasmid and its activity detection
摘要
Abstract
Objective To lay a foundation for further study on the mechanism of Tudor-SN protein in translation . Methods The human Tudor-SN UTR fragments were amplified by PCR from genomic DNA extracted from HeLa cells , and then were inserted into pGL 3-Control expression vector .HeLa cells were co-transfected with the renilla luciferase plasmid and recombinant pGL3-5′UTR plasmid or pGL3-3′UTR plasmid.After 48 hours, luciferase activity was detected . Results Both double enzyme digestion and gene sequencing confirmed the construction of recombinant plasmid was suc -cessful.The luciferase activity was detected after transfection , and the luciferase activity of pGL3-5′UTR plasmid was the best.Conclusion The recombinant plasmids of pGL 3-5′UTR-luciferase and pGL3-3′UTR-luciferase are successfully con-structed, which lays a foundation for further study of regulatory mechanisms of Tudor -SN genetic UTR in translation .关键词
人Tudor-SN蛋白/5′UTR/3′UTR/重组质粒/荧光素酶Key words
human Tudor-SN protein/5′UTR/3′UTR/recombinant plasmid/luciferase分类
生物科学引用本文复制引用
赵亚丽,苏超,甘世虎,任媛媛,高星杰,杨洁,何津岩..人 Tudor-SN UTR 片段荧光素酶报告基因表达质粒的构建及活性检测[J].山东医药,2017,57(4):1-4,4.基金项目
国家杰出青年基金资助项目（31125012）；教育部“创新团队发展计划”（ IRT13085）；国家自然科学基金资助项目（31170830／31370749／31571380）；天津市应用基础与前沿技术研究计划（青年基金项目）（15JCQNJC09900）；天津医科大学青年基金项目（2015KYZQ03）。 （）
