果蝇heix基因启动子的分析及其转录活性鉴定

果蝇heix基因启动子的分析及其转录活性鉴定 Analysis of Drosophila heix Gene Promoter and Characterization of heix Promoter Transcriptional Activity 赵为 ZHAO Wei 1 first-author 夏炎枝 XIA Yanzhi 1 肖菡 XIAO Han 1 夏燕 XIA Yan 1 李欣玥 LI Xinyue 1 红凌 HONG Ling 1 华中科技大学生命科学与技术学院/分子生物物理教育部重点实验室 华中科技大学生命科学与技术学院/分子生物物理教育部重点实验室 College of Life Science and Technology,Huazhong University of Science and Technology/ Key Laboratory of Molecular Biophysics of Ministry of Education College of Life Science and Technology,Huazhong University of Science and Technology/ Key Laboratory of Molecular Biophysics of Ministry of Education 采用生物信息学的方法,综合运用启动子预测工具Promotor Scan1.7和BDGP（neural network pro-moter prediction）分析了目标序列的启动子特征,以果蝇基因组DNA为模板扩增出heix基因启动子候选序列连接至pMD19T载体,构建了荧光素酶报道基因重组质粒pHeixpromoter-Luc,转染果蝇S2细胞表达荧光素酶,并测定了荧光素酶的活性.预测出4个候选的heix基因启动子序列,发现存在ATF、MLTF、c-fos U S5等多种转录因子调控基序;成功构建了pMD19T-Heixpromoter和pHeixpromoter-Luc重组质粒.与肌动蛋白启动子（actinp romoter）相比,pHeixpromoter-Luc表达出的荧光素酶也有较强的活性.本研究找到了果蝇heix基因启动子候选序列,并发现多种转录调控元件,其荧光素酶报告基因实验说明果蝇heix基因启动子与肌动蛋白启动子有相当的转录活性. Through bioinformatic analysis,target sequences of heix gene promoter was analyzed by promoter prediction tools PromotorScan1.7 and BDGP: Neural Network Promoter Prediction.heix gene promoter candidate sequence was amplified using Drosophila genomic DNA as template.Recombinant plasmid pHeixpromoter-Luc was constructed and transfected into Drosophila S2 cells.The luciferase activity of pHeixpromoter-Luc was determined.Four candidates of the heix gene promoter sequences were predicted by bioinformatic softwares.ATF,MLTF,c-fos U S5 and other transcription regulating motifs were found in the candidate sequences.pHeixpromoter-Luc plasmid was constructed successfully and transfected into Drosophila S2 cells.pHeixpromoter-Luc plasmid has strong luciferase activity compared to the actin promoter（actinpromoter）.This study revealed the sequence of Drosophila heix gene promoter and a variety of potential transcriptional regulatory elements,verifying that the predicted heix promoter has transcriptional activity. 启动子 heix基因 荧光素酶 果蝇 promoter heix gene luciferase Drosophila 10.14188/j.1671-8836.2011.01.004 Q78 国家高技术研究发展计划（863）项目（2007AA09Z449）;国家自然科学基金（30971608）;国家创新药物专项基金（2009ZX09301-014）;湖北省自然科学基金（2009CDB074）;中央高校基本科研业务费专项资金（HUST:M2009050）资助项目;武汉市国际科技合作项目（201070934334） 2011-01-01 2021-04-01 1 57