GPC3启动子报告基因载体构建与转录活性分析
投稿时间:2010-04-27    点此下载全文
引用本文:金荣华,陈曦,王美霞,张洪海,孙玉,陈德喜.GPC3启动子报告基因载体构建与转录活性分析[J].医学研究杂志,2010,39(9):47-50
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作者单位
金荣华 首都医科大学附属北京佑安医院药理基地 
陈曦 首都医科大学附属北京佑安医院药理基地 
王美霞 首都医科大学附属北京佑安医院药理基地 
张洪海 首都医科大学附属北京佑安医院感染中心实验室 
孙玉 首都医科大学附属北京佑安医院感染中心实验室 
陈德喜 首都医科大学附属北京佑安医院感染中心实验室 
基金项目:首都医学发展科研基金(2007-2050)
中文摘要:目的构建磷脂酰肌醇蛋白聚糖3(GPC3)启动子荧光素酶报告基因载体,并验证其转录活性。方法以人类基因组DNA为模板,PCR扩增GPC3启动子基因片段,克隆到pGL3-Basic荧光素酶报告基因载体上,通过转染HepG2细胞检测荧光素酶活性来验证GPC3启动子的转录活性。结果GPC3基本启动子和全长启动子分别成功构建到pGL3-Basic荧光素酶报告基因载体中,GPC3基本启动子具有很强的转录活性,而全长启动子的转录活性比基本启动子高4倍以上。结论成功构建了GPC3启动子荧光素酶报告基因载体,为研究GPC3转录调控提供了重要手段。
中文关键词:GPC3  启动子  报告基因  荧光素酶
 
Construction of GPC3 Promoter Reporter Gene Vector and Analysis of its Transcriptional Activity
Abstract:ObjectiveTo construct glypican-3 (GPC3) promoter luciferase reporter gene vector, and to analyze its transcriptional activity. MethodsThe human genomic DNA was extracted as template.GPC3 promoter gene fragment was amplified and cloned into PGL3-Basic luciferase reporter gene vector. After HepG2 cells being transfected,the transcriptional activity of GPC3 promoter was verified through the detection of luciferase activity. ResultsBasal GPC3 promoter and full-length GPC3 promoter were successfully constructed to PGL3-basic luciferase reporter gene vector.GPC3 basal promoter had a strong transcriptional activity, while that of the full-length promoter was 4 times higher. ConclusionGPC3 promoter luciferase reporter gene vector was successfully constructed.
keywords:Glypican-3  Promoter  Reporter gene  Luciferase
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