hhlim基因转录调控区域鉴定及表达调控的初步研究  

Identification of hhlim Gene Regulatory Region and Study of hhlim Gene Expression Regulation

作　　者：郑斌[1] 温进坤[2] 韩梅[1] 周爱儒[3] 

机构地区：[1]河北医科大学基础医学研究所,石家庄050017 [2]河北医科大学基础医学研究所生物化学与分子生物学研究室 [3]北京大学医学部,生物化学与分子生物学系,北京100083

出　　处：《生物化学与生物物理进展》2002年第6期910-914,共5页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展规划项目 (973 )基金资助(G2 0 0 0 0 5 690 5 Z)~~

摘　　要：为了研究hhlim基因表达调控机制 ,对该基因 5′上游 - 2 5 37bp序列从 5′端依次进行缺失后 ,利用荧光素酶报告基因检测各种不同长度片段在C2C12细胞中驱动荧光素酶表达的活性 .结果表明 ,在hhlim基因 5′上游- 2 5 37～ - 15 37bp之间存在负调控元件 ,在 - 2 5 3～ - 15 7bp之间含有增强子样序列 .用含有增强子样序列的DNA片段做探针 ,对未分化型和分化型C2C12细胞的核蛋白进行电泳迁移率改变分析 (electrophoreticmopilityshiftassay ,EMSA) .分析的结果显示 ,两种表型细胞中的核蛋白与探针结合所形成滞后带的谱型明显不同 .结果还发现内皮素 1(ET 1)和碱性成纤维细胞生长因子 (bFGF)既能显著诱导C2C12细胞对hhlim的表达 ,也能刺激含增强子样序列的调控区域所驱动的报告基因表达 .提示hhlim基因转录起始点至 - 2 5 37bp的区域内含有负调控元件及增强子样序列 ,该基因的表达受ETIn order to study the mechanism of hhlim gene transcriptional regulation, a series of deleted fragments of 5′ flanking region extending from +16 to -2 537 bp were subcloned into the pGL3 Basic vector respectively to identify the specific functional regions by detecting the luciferase activities. The results indicated that there was a silencer in the distal region of -2 537～-1 537 bp and an enhancer in proximal fragment of -253 ～-157 bp, respectively. Electrophoretic mobility shift assay showed that the patterns of shifted bands were different between the nuclear protein from differentiated C2C12 myotubes and undifferentiated C2C12 myoblasts when they bound to the region including the region of -253～-157 bp of hhlim gene. In addition, the results also showed that ET 1 and bFGF could not only significantly induce the hhlim gene expression in C2C12 cells but also activate the luciferase gene transcription promoted by -253～-157 bp regulatory region. It was suggested that hhlim gene was regulated by ET 1 and bFGF.

关 键 词：hhlim基因转录 调控区域 鉴定 表达调控 调控序列 成肌细胞 

分 类 号：Q75[生物学—分子生物学]