人TSLC1基因核心启动子的克隆和鉴定

基础医学与临床 ›› 2010, Vol. 30 ›› Issue (4): 406-410.

人TSLC1基因核心启动子的克隆和鉴定

高静申林明树红

北京大学临床肿瘤学院北京肿瘤医院暨北京市肿瘤防治研究所中国医学科学院北京协和医学院基础医学研究所卫生部北京医院呼吸科

收稿日期:2009-12-11 修回日期:2009-12-23 出版日期:2010-04-05 发布日期:2010-04-05
通讯作者: 明树红

Clone and identification of the core promoter of human TSLC1 gene

Jing GAO, Lin SHEN, Shu-hong MING

Peking University School of Oncology, Beijing Cancer Hospital & Institute Dep. of Respiratory Beijing Hospital Ministry of Health

Received:2009-12-11 Revised:2009-12-23 Online:2010-04-05 Published:2010-04-05
Contact: Shu-hong MING

摘要/Abstract

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～-329bp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～-329bp区域可能为基因的核心启动子区。

关键词: TSLC1, 核心启动子, 克隆, 鉴定

Abstract: Objective To clone and identify the core promoter of human TSLC1 used for carrying out the study 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 relative activities of different fragments in A549 and NCI-H446 cells were detected by dual-luciferase assay after transient transfection, and then the core promoter of TSLC1 was identified. Results Among the different constructs, the fragment of -68～-329bp 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～-329bp located in the upstream of translation start site of TSLC1 may be the core promoter region.

Key words: TSLC1, core promoter, clone, identify

高静申林明树红. 人TSLC1基因核心启动子的克隆和鉴定[J]. 基础医学与临床, 2010, 30(4): 406-410.

Jing GAO; Lin SHEN; Shu-hong MING. Clone and identification of the core promoter of human TSLC1 gene[J]. Basic & Clinical Medicine, 2010, 30(4): 406-410.

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