人β1整合素启动子核心启动序列初步分析  

The Preliminary Analysis of Core Sequences at Human Integrin β1 Promoter

作　　者：王秀亮[1] 陈卫[1] 梁光萍[1] 陈建[1] 苏踊跃 杨程[1] 罗向东[1] 

机构地区：[1]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 [2]昆明军区总医院,云南昆明650000

出　　处：《现代生物医学进展》2009年第1期50-52,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30730093)

摘　　要：目的:分析人β1整合素在HaCat细胞中核心启动片段。方法:以本室构建的含整合素β1全长启动子1745bp基因序列(-1745bp～+11bp)的重组载体pGL3-1756为模板,用含有酶切位点的特异性引物扩增出整合素β1启动子-845bp～-304bp间基因序列,克隆至荧光素酶表达载体pGL3basic,构建含正确目的基因重组载体pGL3-542。用pGL3-1756、和pGL3-542质粒转染人表皮细胞株HaCat进行活性分析。结果:酶切及序列测定表明,克隆后插入pGL3basic中的启动子片段与GenBank DNA序列数据库对比分析序列一致,且插入方向正确。在人永生化表皮细胞株(HaCat细胞)中构建的pGL3-542载体具有很强的启动活性。结论:成功构建了整合素β1远端启动子542bp片段载体,在HaCat细胞中具有非常强的启动活性。人整合素β1整合素启动子核心启动序列可能位于-845bp～-304bp。Objective： To analyze the core fragment of integrin betal in HaCat cells. Methods： pGL3-1756, a constructed vector containing the human integrin beta 1 promoter （1756 bp）, was used as template for amplification of the vector, which was named pGL3-542. HaCat cells were transfected with pGL3-1756, promoter sequence at the distal part （-845 bp～-304 bp）. Then the amplified sequence was cloned into pGL3 basic pGL3-542, respectively, for the analysis of promoter activity, and pGL3-basic was used as negative controls. Results： The results of enzyme digestion and sequencing identified the correction of constructed vector, pGL3-542 exhibited very strong activity in HaCat cells. Conclusion： The reporter gene vector containing 542 base pairs at the distal part ofintegrin beta 1 promoter is constructed successfully, with strong activity in HaCat cells. The core sequences at human integrin β1 promoter may be located in -845 bp ～-304 bp.

关 键 词：整合素Β1 启动子 载体 

分 类 号：Q78[生物学—分子生物学] Q26