| 冉莉,涂晶,肖新华,钟警,洪涛,文格波.小鼠PGC-1α基因启动子的转录调控功能分析.[J].中南医学科学杂志.,2013,41(6):546-550. |
| 小鼠PGC-1α基因启动子的转录调控功能分析 |
| Analyzing the Transcription Regulation Function ofPGC-1α Gene Promoter in Mouse |
| 投稿时间：2013-08-27 |
| DOI： |
| 中文关键词: PGC-1α 启动子 转录调控 小鼠 肝脏 |
| 英文关键词:PGC-1α promoter transcriptional regulation mouse liver |
| 基金项目:国家自然基金项目（30900707）. |
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| 中文摘要: |
| 目的克隆小鼠PGC-1α基因的启动子并分析其转录调控模式。方法设计引物，以小鼠肝脏组织DNA 为模板，PCR扩增PGC-1α 基因启动子区并克隆入荧光素酶报告基因表达载体pGL3-Basic 中，构建具有PGC-1α启动子转录活性的报告质粒pGL3-PGC-1α-Luc。通过特异性引物，引入替换碱基突变相关转录因子调控模序，构建点突变融合载体,并将其转染1c1c7 及c2c12 细胞系，检测荧光素酶的表达情况。结果构建的pGL3-PGC-1α-Luc 系列报告质粒经过酶切鉴定及DNA 测序分析都显示正确；PGC-1α基因启动子区域（-2533~+78）具有较强的转录活性；突变失活PGC-1α基因启动子区域中的CRE及IRE反应原件导致转录活性明显降低。结论成功构建小鼠PGC-1α基因启动子报告质粒，PGC-1α 基因上游区域（-2533~+78）具有较强的启动子活性；启动子区域中的CRE和IRE反应原件为主要转录调控位点。 |
| 英文摘要: |
| ObjectiveTo clone the promoter of mouse PPARg co-activator 1α(PGC-1α)gene and analyze its transcriptional regulation mode.MethodsTake the mice liver tissue DNA as a template and the primers were designed,then this promoter was cloned by PCR and transfected it into the luciferase reporter gene expression vector pGL3-Basic.The report plasmid pGL3-PGC-1 a-Luc contained transcription activity of PGC-1 promoter was built.Building the point mutation fusion vector by inserting specific primers the substitution mutation related transcription factor regulatory motif.Transfected the point mutation fusion vector to the 1c1c7 and C2C12 cell and tested its expression by luciferase.ResultsThe results showed that the construction of the pGL3-PGC-1α-Luc series of reporter plasmids and DNA sequencing analysis were correct,and promoter region of PGC-1α gene(-2533~+78)had a strong transcriptional activity;The CRE and IRE response element in mutational inactivation of PGC-1α gene promoter decreased the transcription activity.ConclusionsSuccessful constructed the mouse PGC-1 gene promoter reporter plasmid and confirmed the upstream region(-2533~+78)of the PGC-1α gene has a strong promoter activity.The CRE and IRE response element in the area of promoter is the main transcriptional regulatory sites. |
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