EOLA1基因启动子序列的鉴定  

Identification of EOLA1 gene promoter sequence

作　　者：梁自文[1] 周广举[1] 杨宗城[2] 陈建[2] 陈渝[2] 

机构地区：[1]第三军医大学西南医院内分泌科,重庆400038 [2]第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室,重庆400038

出　　处：《第三军医大学学报》2009年第23期2309-2311,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金(30200101;30670838)~~

摘　　要：目的鉴定EOLA1基因启动子序列,为深入研究EOLA1的转录调控机制奠定基础。方法通过PCR反应进一步缺失突变EOLA1基因5′侧翼区-1672～+51(1723bp)序列,将产物插入到含荧光素酶报告基因的载体pGL3-Basic中,转染ECV304细胞,瞬时表达后测定荧光素酶活性。采用生物信息学分析该1723bp片段可能存在的转录因子结合位点。结果缺失到785bp片段仍具有启动子活性,进一步缺失到717bp片段后活性消失,从而确定启动子位于-738～-676bp序列,其附近含Sp1和Myf顺式作用元件。结论确定了EOLA1启动子范围和转录因子结合位点。Objective To identify the promoter sequence of endothelial-overexpressed lipopolysaccharide- associated factor 1 （EOLA1） gene and to elucidate the molecular mechanisms controlling EOLA1 expression. Methods A DNA fragment containing 1 723 bp 5＇ upstream of the EOLA1 gene and the transcription start site was generated by polymerase chain reaction and then cloned into a luciferase reporter gene vector, pGL3-basic. The relative luciferase activities driven by this 5 ＇-upstream fragment and a series of deletion mutants were measured in transiently transfected human ECV304 cells, respectively. At last, the 1 723 bp upstream of the EOLA1 gene was analyzed online with Cluster Buster. Results A fragment 785 bp upstream of the EOLA1 coding region was sufficient to promote transcription. Further deletion analysis of the 785 bp fragment indicated that a 68 bp element from -738 to -676 was important for EOLA1 transcription in ECV304 cells. The 1723 bp sequence contains binding sites for Spl and Myf. Conclusion We map the EOLA1 promoter by deletion analysis and reveal that the proximal region （ -738 to -676 bp） , which contains binding sites for Spl and Myf, is essential for human EOLA1 promoter activity in ECV304 cells.

关 键 词：EOLA1 启动子 转录因子 报告基因 

分 类 号：R394-33[医药卫生—医学遗传学] R394.3[医药卫生—基础医学]