小鼠TLE4基因启动子克隆及分析  

Cloning and Analyzing the Promoter of Mouse TLE4 Gene

作　　者：赵黎黎[1,2] 牟玉莲[2] 崔文涛[2] 杨述林[2] 唐中林[2] 李勇 赵为民[2] 刘娣 李奎[2] 

机构地区：[1]东北农业大学动物科学技术学院,哈尔滨150030 [2]中国农业科学院北京畜牧兽医研究所畜禽遗传资源与利用农业部重点开放实验室,北京100193

出　　处：《畜牧兽医学报》2010年第3期274-278,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30871775;30830080);国家863计划和重大基础研究计划资助(2006AA10Z135;2006CB102105)

摘　　要：本研究旨在初步对小鼠TLE4基因的转录调控机制进行探讨。利用PCR方法扩增TLE4基因5′上游启动区2 849 bp(-2 521 bp^+327 bp)的片段,然后通过步移缺失获得了7段长度不等的启动子片段并分别克隆到荧光素酶(LUC)报告基因表达质粒中。通过双荧光素酶报告活性分析检测TLE4基因启动子区不同长度片段在小鼠畸胎瘤细胞(F9)及小鼠胚胎干细胞(ES)中瞬时转染后的活性。2种细胞的检测结果显示,在TLE4基因启动子区(-2 521 bp^-2 137 bp)存在负性调控元件,而在启动区(-2 137 bp^-1 794 bp)活性最强。对TLE4基因启动区(-2 137 bp^-1 794 bp)进一步缺失分析发现在该基因启动区(-2 027 bp^-1 927 bp)活性较强,分析预测该序列含有一个功能性的(HSF)的结合位点。结果推测HSF对TLE4基因的表达调控及功能行使具有重要作用。The preliminary objective of the study was to investigate the transcriptional regulatory mechanisms of mouse TLE4.5＇upstream promoter sequences （spanned from --2 521 bp to ＋327 bp） of mouse TLE4 gene were amplified by PCR. Seven promoter fragments with different length were obtained by walking deletion and then cloned into luciferase report gene expression vectors. The vector expression activities were determined by transfection of the mouse F9 teratoma cells and the mouse embryonic stern cells with the constructed dual-luciferase vectors. The experiment results indicated that negative regulation elements were localized within the promotor region from --2 521 bp to --2 137 bp, and further deletion analysis indicated that the region from --2 027 bp to -1 927 bp in TLE4 gene promotor showed higher activity than other regions. In addition, a functional HSF regulation element was identified in this region. The results inferred that the HSF played an important role in the regulation of TLE4 gene expression and its functional performance.

关 键 词：TLE4基因 启动子 HSF 调控序列 

分 类 号：Q523.8[生物学—生物化学]