小麦非特异转脂蛋白TaLTP1启动子区域顺式作用元件鉴定及转录调控机制研究

项目名称： 小麦非特异转脂蛋白TaLTP1启动子区域顺式作用元件鉴定及转录调控机制研究

项目编号： No.31200982

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 王宏伟

作者单位： 山东农业大学

项目金额： 23万元

中文摘要： 很多非特异性转脂蛋白（nsLTP）表达在植物地上组织表皮中，被认为参与了一系列表皮相关的生理过程。目前，对植物地上组织表皮，尤其是铺列细胞的转录调控机制的理解非常匮乏，因此本项目拟以TaLTP1 启动子为研究对象，首先进行5'端序列续减分析确定启动子功能区域，然后通过生物信息学、凝胶迁移和DNA顺序突变方法配合运用，准确鉴定功能顺式作用元件，分析顺式调节模式及其进化路径。为进一步揭示转录调控激活机制，利用酵母单杂交和酵母展示cDNA文库技术分离潜在的转录因子，研究其在烟叶中的亚细胞定位及与调控元件相互作用的机制。本研究通过阐明TaLTP1启动子针对地上组织表皮表达的转录调控机制，为进一步理解植物地上组织表皮细胞调控基因转录表达路径奠定基础。另外，利用酵母展示cDNA文库技术对与调控元件特异结合的转录因子进行筛选，将为分子生物学领域有效分离转录因子开辟新的模式方法。

中文关键词： 小麦；非特异转脂蛋白；转录调控；表皮；调控元件

英文摘要： Many non-specific lipid transfer protein (nsLTP) genes were found to be expressed on epidermis of plant aerial tissue, which were thought to participate multiple functions related with epidermal cells including in lipid metabolism, cuticle biosynthesis, plant growth, or biotic/abiotic stress resistance. Currently, little is known about plant differentiation of ground epidermal cells and its transcriptional regulation. Here, we attempt to analyze the cis-regulation of TaLTP1 promoter and transactivation mechanisms. Firstly, a series of TaLTP1 promoter deletion mutants are transformed into Arabidopsis for activity characterization and narrowing down the functional promoter fragments. Then, the identification of cis-elements specific to shoot epidermis will be identified by employing the bioinformatics tools, electrophoretic mobility shift assay (EMSA) and promoter mutatants in transgenic Arabidopsis, followed by analysis of the the cis-regulatory mode and evolutionary track. In order to demonstrate the transactivation mechanisms, the potential transcription factors will be isolated by yeast one hybrid and yeast surface display systems. The DNA binding specificity of isolated transcription factors will be analyzed by EMSA and the subcellular localization will be examined in transgenic tobacco by fusion with EGFP. B

英文关键词： wheat；nsLTP；transcription regulation；epidermis；cis-element