银杏MECPs基因启动子克隆及其植物表达载体的构建  

Cloning of MECPs Gene Promoter from Ginkgo biloba and Construction of Plant Expression Vector

作　　者：袁红慧[1,2,3] 程华[2,3] 李琳玲[2,3] 许锋 程水源[1,2,3] 

机构地区：[1]武汉工程大学化工与制药学院,湖北武汉430073 [2]经济林木种质改良与资源综合利用湖北省重点实验室,湖北黄冈438000 [3]黄冈师范学院化学与生命科学学院,湖北黄冈438000

出　　处：《贵州农业科学》2013年第5期10-15,共6页Guizhou Agricultural Sciences

基　　金：国家自然科学基金项目"银杏叶黄酮关键基因GbPAL1与GbCHS2启动子的克隆与功能研究";"两个WD40转录因子对银杏类黄酮生物合成调控的研究"(30971974;31270717);湖北省自然科学基金创新群体项目"银杏萜内酯合成途径中两个关键基因DXS与LPS启动子的分离及其调控研究"(2011CDA117)

摘　　要：为深入研究银杏MECPs基因启动子的结构及功能特点,以前期获得的银杏MECPs基因的cDNA序列为模板,采用染色体步移技术(Genome Walking),从银杏基因组中扩增到一段GbMECPs基因启动子序列。结果表明:该启动子片段长度约为744bp,含有多个基本顺式作用元件TATA和CAAT-box及典型的光调节元件GT-1motif(GRWAAW)和Circadian box(CAANNNNATC)等。进一步分析发现,该调控序列还包含多个对生长素、赤霉素、乙烯利、细胞分裂素、脱落酸等各种激素应答相关的调控元件和MYB、MYC转录因子介导的多种逆境胁迫诱导元件。将克隆的GbMECPs启动子替换质粒pBI121中的35S启动子,成功构建了由GbMECPs启动子驱动GUS报告基因的植物表达载体pBI121+GbMECPs。In order to learn more about the substructure and function of the GbMECPs, the authors designed primers according to the cDNA sequence of MECPs gene and obtained MECPs gene promoter from tall rescue genomic DNA by genome walking. The results showed that the GbMECPs promoter region was 744bp long and contained several TATA and CAAT boxes, as well as some typical light- regulated elements, such as GT-1 motif （GRWAAW）, Circadian box （CAANNNNATC） et al. Furthermore, there were various kinds of hormone-regulated elements that response to auxin, gibberellin, ethephon, cytokinins and abscisic acid, and some elements related to plant stress responses that mediated by MYB, MYC transcription factor. By replacing the 35S promoter, MECPs promoter was inserted upstream of the Gus reporter gene in plasmid pBI121, and the plant expression vector pBI121＋GbMECPs was constructed.

关 键 词：银杏 染色体步移 2-C-甲基赤藓醇-2 4-环焦磷酸合成酶基因 调控元件 植物表达载体 

分 类 号：Q784[生物学—分子生物学]