利用补丁甲基化技术构建特殊报告载体检测DNA甲基化对FCER1G基因启动子的调控活性  

Construction of special reporter to detect DNA methylation regulatory activity in FCER1G gene promoter through patch-methylation

作　　者：梁云生[1] 赵明[1] 梁功平[1] 尹恒[1] 陆前进[1] 

机构地区：[1]中南大学湘雅二医院皮肤科,医学表观基因组学湖南省重点实验室,长沙410011

出　　处：《中南大学学报（医学版）》2013年第2期120-124,共5页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金青年项目(30800993/H1103);国家重点基础研究计划(973计划)(2009CB825605)~~

摘　　要：目的:构建一种特殊的荧光素报告载体检测DNA甲基化对FCER1G基因启动子调控序列的调控活性。方法:用补丁甲基化技术构建特殊的含完全甲基化和模拟甲基化的FCER1G启动子调控序列的PGL-3荧光素报告载体;通过比较完全甲基化与模拟甲基化荧光素报告载体活性来检测DNA甲基化对FCERG基因启动子调控序列的调控活性。结果:成功构建特殊的完全甲基化与模拟甲基化FCER1G启动子调控序列荧光素报告载体;并检测出完全甲基化与模拟甲基化荧光素报告载体活性比为(0.36±0.07):1(P<0.001)。结论:FCER1G启动子调控序列是甲基化敏感位点,FCER1G启动子受DNA甲基化调控。Objective： To construct a special luciferase reporter to detect DNA methylation regulatory activity in FCER1 G gene promoter regulatory element. Methods： We constructed special full and mock methylated FCER1G gene promoter regulatory luciferase reporters by patch-methylation, and detected DNA methylation regulatory activity by comparing the luciferase activity of full-methylated luciferase reporters with mock-methylated reporters. Results： We successfully constructed the full and mock methylated FCERIG gene promoter regulatory luciferase reporters. The ratio of luciferase activity between the full methylated and themock methylated was （0.36±0.07）： 1 （P〈0.001）. Conclusion： FCER1G promoter activity is methylation-sensitive and is regulated by DNA methylation.

关 键 词：补丁甲基化 FCER1G基因 启动子调控序列 DNA甲基化 

分 类 号：R440[医药卫生—诊断学]