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
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。
[Construction of special reporter to detect DNA methylation regulatory activity in FCER1G gene promoter through patch-methylation]
OBJECTIVE: To construct a special luciferase reporter to detect DNA methylation regulatory activity in FCER1G 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 FCER1G gene promoter regulatory luciferase reporters. The ratio of luciferase activity between the full methylated and the mock methylated was (0.36±0.07):1 (P<0.001). CONCLUSION: FCER1G promoter activity is methylation-sensitive and is regulated by DNA methylation.
pmid: 23456064 Zhong Nan Da Xue Xue Bao Yi Xue Ban 影响因子: 0.0 发表日期: 20130201 官网 免费下载 全文下载
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