人TSLC1基因核心启动子的克隆和鉴定  

Cloning and identification of the core promoter of human TSLC1 gene

作　　者：高静[1,2,3] 申林[4] 明树红[5] 

机构地区：[1]北京大学临床肿瘤学院 [2]北京肿瘤医院 [3]北京市肿瘤防治研究所消化肿瘤内科恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142 [4]中国医学科学院北京协和医学院基础医学研究所医学分子生物学国家重点实验室,北京100005 [5]卫生部北京医院呼吸科,北京100730

出　　处：《基础医学与临床》2010年第4期406-410,共5页Basic and Clinical Medicine

摘　　要：目的克隆和鉴定人TSLC1基因的核心启动子区,用于开展其转录调控机制的研究。方法用PCR方法从人基因组DNA中扩增出TSLC1基因翻译起始位点上游一系列大小不等的片段,连入pGL3-Basic荧光素酶报告基因载体中。通过瞬时转染A549及NCI-H446细胞,检测细胞裂解液中的双荧光素酶活性,确定TSLC1基因的核心启动子区。结果在人TSLC1基因启动子区的分段克隆中,ATG上游-68～-329 bp片段在A549及NCI-H446细胞中均具有很强的启动活性,对TSLC1的转录起重要作用。结论人TSLC1基因翻译起始位点ATG上游-68～-329 bp区域可能为基因的核心启动子区。Objective To clone and to identify the core promoter of human TSLC1 used for exploring of transcription regulatory mechanism.Methods A series of different fragments located in the upstream of translation start site of TSLC1 were amplified from human genomic DNA by PCR,and then constructed into pGL3-Basic luciferase reporter vector.The activity of different fragments in A549 and NCI-H446 cells was examined by a dual-luciferase assay after transient transfection,and then the core promoter of TSLC1 was identified.ResultsAmong the different constructs,the fragment of-68～-329 bp located in the upstream of ATG showed the strong activity both in A549 cells and NCI-H446 cells,which played an important role in the transcription of TSLC1.Conclusion The fragment of-68～-329 bp located in the upstream of translation start site of TSLC1 might be the core promoter region.

关 键 词：TSLC1 核心启动子 克隆 鉴定 

分 类 号：R73-3[医药卫生—肿瘤] R346[医药卫生—临床医学]