Regulation of Gene Expression by Ethylene

乙烯对基因表达的调控

• 批准号: 9004353
• 负责人: Robert Fischer
• 金额: $ 25.5万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9004353&HistoricalAwards=false
• 关键词: Regulation; Gene; Expression; Ethylene

The long term objective of this research is to understand the molecular and cellular processes by which ethylene controls fruit ripening and gene expression. The model system is Lycopersicon esculentum (tomato). By analyzing the effect of promoter deletions on gene expression in fruit from transformed plants, and in a recently developed microprojectile transient assay system, 5'-flanking regions that control gene expression in a positive and negative manner have been identified. Sites that react with developmentally regulated DNA-binding activities have been delineated, and a clone encoding a DNA-binding protein has been isolated. The specific goal is to elucidate the mechanisms of ethylene action at the gene level. Ethylene-inducible promoter fine structure will be analyzed by determining the effect of promoter mutations on gene expression in a transient gene expression system and in transgenic fruit. Specific mutations will include deletion of sites that react with DNA binding proteins, and alterations of binding site position and orientation within the promoter. Factors that regulate ethylene inducible gene expression will be studied by cloning DNA binding proteins that bind to critical DNA regulatory sequences. The cloned gene will be used to generate specific antibodies and these tools will be used to determine the mechanism that regulates DNA binding activity. DNA binding protein function will be elucidated genetically by analyzing the phenotype of transgenic plants with reduced levels of DNA binding activity due to the expression of an antisense DNA binding protein gene. The interaction of the DNA binding protein with ethylene and/or other nuclear proteins will be analyzed. %%% The plant hormone, ethylene, controls fruit ripening. This project is an investigation, at the molecular and genetic level, of how that control is carried out.

这项研究的长期目标是了解乙烯控制果实成熟和基因表达的分子和细胞过程。该模型系统是 Lycopersicon esculentum（番茄）。通过分析启动子缺失对转化植物果实中基因表达的影响，并在最近开发的微弹瞬时测定系统中，已经鉴定出以正向和负向方式控制基因表达的5'侧翼区域。与发育调节的 DNA 结合活性发生反应的位点已经被描绘出来，并且编码 DNA 结合蛋白的克隆也已经被分离出来。具体目标是阐明乙烯在基因水平上的作用机制。通过确定启动子突变对瞬时基因表达系统和转基因水果中基因表达的影响，将分析乙烯诱导型启动子的精细结构。特定突变包括删除与 DNA 结合蛋白反应的位点，以及改变启动子内的结合位点位置和方向。将通过克隆与关键 DNA 调控序列结合的 DNA 结合蛋白来研究调控乙烯诱导基因表达的因素。克隆的基因将用于产生特异性抗体，这些工具将用于确定调节 DNA 结合活性的机制。 DNA结合蛋白的功能将通过分析由于反义DNA结合蛋白基因的表达而导致DNA结合活性水平降低的转基因植物的表型来从遗传学上阐明。将分析DNA结合蛋白与乙烯和/或其他核蛋白的相互作用。 %%% 植物激素乙烯控制果实成熟。该项目是在分子和基因水平上研究如何进行这种控制的。