Purification and Cloning of Elicitor Binding Protein(s) from Soybean
大豆中诱导子结合蛋白的纯化和克隆
基本信息
- 批准号:9206882
- 负责人:
- 金额:$ 54.7万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Continuing Grant
- 财政年份:1993
- 资助国家:美国
- 起止时间:1993-02-15 至 1997-01-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The long term goal of this research is to understand how plant cells perceive and respond to extracellular signals. The system being studied is the induction of plant defense responses, phytoalexin accumulation in soybean (Glycine max) induced by oligosaccharides (elicitors) originating from the mycelial wall of a phytopathogenic fungus, Phytophthora megasperma f. sp. glycinea. Considerable information has been obtained about the structure of an elicitor (a branched hepta-beta-glucoside) derived from mycelial wall glucans and about the identity and regulation of elicitor- induced genes encoding enzymes required for the biosynthesis of the phytoalexins. However, little is known about the mechanisms by which plant cells perceive the elicitor, or how that signal is transmitted to the cell nucleus to initiate changes in gene expression. The research described in this proposal focuses on the first step in the elicitor-stimulated signal transduction pathway, that is, the recognition of a hepta-beta-glucoside elicitor by a plasma-membrane localized receptor. The proposed research has three major goals. The first goal is the identification, purification, and characterization of hepta-beta-glucoside elicitor binding protein(s) (EBPs) from soybean root microsomal membranes. Evidence has already been obtained that a single class of specific, high-affinity (dissociation constant of 1 nM) EBPs exist in soybean root microsomes originating from the plasma membrane. These EBPs have been successfully solubilized from the membranes using detergents, and the solubilized EBPs retain their high affinity and specificity for the elicitor. The number and identity of the proteins that have hepta-beta-glucoside elicitor binding activity will be established by photo-affinity labeling. The EBPs will be purified by affinity chromatography and antibodies against the binding protein(s) will be prepared. The second goal is the isolation and characterization of cDNA sequences that encode EBPs. Clones carrying EPB sequences will be identified by screening cDNA expression libraries with radiolabeled hepta-beta-glucoside elicitor. Alternatively, partial protein sequences obtained from the purified EBPs will be used to prepare synthetic nucleic acid probes with which to screen cDNA libraries. These clones and their derived sequences will be used to identify possible structural and functional domains in the EBPs that might relate to their role in signal transduction. The third goal, which will be undertaken if time permits, is to demonstrate that the EBPs identified in these studies are physiological receptor(s) for the elicitor. Sequence comparisons and expression of truncated EBP genes will be used to identify and delineate functional domains of the EBPs. Transgenic expression of wild-type and mutant EBPs in plant cells that do not contain endogenous EBPs, tissue-specific localization of EBPs in relation to localization of plant defense responses, and determination of the effect of anti-sense EPB sequences on the expression of hepta-beta-glucoside-induced plant defense responses will be undertaken to demonstrate the physiological significance of the EBPs in elicitor-mediated signal transduction. %%% The long term goal of this research is to understand how plant cells perceive and respond to extracellular signals. The system being studied is the induction of a defense response in soybean plants. The defense response is triggered by exposure of the plant to certain carbohydrates (elicitors) which originate from the filament wall of a pathogenic fungus which specifically infects the soybean. In response to the elicitors the soybean plant synthesizes and accumulates protective chemicals known as phytoalexins. The more immediate goals of this project are to identify and characterize the plant receptors for the elicitors. Detailed characterization of the elicitor receptor will be an important step toward understanding of the signal transduction pathway by which plants detect and respond to pathogens.
这项研究的长期目标是了解植物 细胞感知并响应细胞外信号。 系统 正在研究的是植物防御反应的诱导, 诱导大豆(Glycine max)植保素积累 低聚糖(激发子)来源于菌丝体壁 一种植物病原真菌,大种子疫霉Phytophthora megasperma f.大豆根瘤菌 已经获得了关于以下结构的大量信息: 一种从菌丝体中提取的激发子(一种分支的七-β-葡糖苷) 壁葡聚糖和关于诱导子的身份和调节, 诱导的基因编码生物合成所需的酶, 植物抗毒素 然而,很少有人知道的机制, 哪些植物细胞感知到了激发子,或者这个信号是如何 传递到细胞核引发基因变化 表情 本提案中所述的研究重点是 激发子刺激的信号转导途径的第一步, 也就是说,七-β-葡糖苷诱导子被 质膜定位受体。 拟议的研究已 三大目标。 第一个目标是识别, 七-β-葡萄糖苷诱导子的分离纯化及性质研究 大豆根微粒体膜结合蛋白(EBP)。 已经获得的证据表明,一个特定的, 大豆中存在高亲和力(解离常数为1 nM)EBP 起源于质膜的根微粒体。 这些EBPs 已经成功地从膜上溶解, 洗涤剂,溶解的EBP保持其高亲和力, 对激发子的专一性。 的数量和身份 具有七-β-葡糖苷诱导子结合活性的蛋白质 将通过光亲和标记来建立。 EBP将在 通过亲和色谱法纯化,并制备抗 将制备结合蛋白。 第二个目标是 编码EBP cDNA序列的分离和表征。 携带EPB序列的克隆将通过筛选cDNA来鉴定 具有放射性标记的七-β-葡糖苷的表达文库 激发子。 或者,可将从以下获得的部分蛋白质序列: 纯化EBP将用于制备合成核酸 探针筛选cDNA文库。 这些克隆人和他们 衍生序列将用于鉴定可能的结构和 EBP中的功能结构域可能与它们在以下方面的作用有关: 信号转导 第三个目标将在以下情况下实现: 时间允许,是为了证明这些EBP中确定的 研究是激发子的生理受体。 序列 截短的EBP基因的比较和表达将用于 鉴定和描绘EBP的功能结构域。 转基因 在植物细胞中表达野生型和突变型EBP, 含有内源性EBPs,EBPs的组织特异性定位, 与植物防御反应定位的关系,以及 测定反义EPB序列对细胞增殖的影响 七-β-葡萄糖苷诱导的植物防御反应的表达 将进行演示的生理意义, 激发子介导的信号转导中的EBP。 %%% 这项研究的长期目标是了解植物 细胞感知并响应细胞外信号。 系统 正在研究的是诱导大豆的防御反应, 植物 防御反应是由植物暴露在 某些碳水化合物(激发子),来源于 一种致病真菌的丝状壁,专门感染 黄豆 大豆植株对诱导剂的反应 合成并积累保护性化学物质, 植物抗毒素 这个项目的更直接的目标是 鉴定和表征激发子的植物受体。 激发子受体的详细表征将是 这是理解信号转导的重要一步 植物检测和应对病原体的途径。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Michael Hahn其他文献
Evaluating the Meaning of Answers to Reading Comprehension Questions: A Semantics-Based Approach
评估阅读理解问题答案的含义:基于语义的方法
- DOI:
- 发表时间:
2012 - 期刊:
- 影响因子:0
- 作者:
Michael Hahn;Walt Detmar Meurers - 通讯作者:
Walt Detmar Meurers
奥田聖應先生頌寿記念インド学仏教学論集(担当論文題目「ダルモーッタラ著『ApohaprakaraNa』の冒頭偈について」)
奥田清夫纪念印度学与佛教论文集(论文标题:《论法陀罗《阿波哈普拉卡拉那》》的开篇诗)
- DOI:
- 发表时间:
2014 - 期刊:
- 影响因子:0
- 作者:
Lambert Schmithausen;Michael Hahn;原實;荒巻典俊;島田外志夫;他85名 - 通讯作者:
他85名
The Effect of Different Annealing Conditions on the Anisotropy of the Fracture Toughness of Ti-6Al-4V
- DOI:
10.1007/s11665-019-04449-6 - 发表时间:
2019-11-14 - 期刊:
- 影响因子:2.000
- 作者:
Alfredo Gutierrez;Michael Hahn;Yong-Jun Li;Abi Dehbozorgi;William Hohorst;Michael Schwartz;Jacob Orlita;Ye Thura Hein;Nelson Guanzon;Xiaodong Sun;Omar S. Es-Said - 通讯作者:
Omar S. Es-Said
Intraosseous lymphocytic infiltrates after hip resurfacing arthroplasty
- DOI:
10.1007/s00428-009-0745-7 - 发表时间:
2009-02-19 - 期刊:
- 影响因子:3.100
- 作者:
Jozef Zustin;Michael Amling;Matthias Krause;Stefan Breer;Michael Hahn;Michael M. Morlock;Wolfgang Rüther;Guido Sauter - 通讯作者:
Guido Sauter
INTEGRATION OF SENTINEL-2 AND LANDSAT-8 DATA FOR SURFACE REFLECTANCE TIME-SERIES ANALYSIS
整合 Sentinel-2 和 Landsat-8 数据进行表面反射率时间序列分析
- DOI:
- 发表时间:
2019 - 期刊:
- 影响因子:0
- 作者:
G. Berdou;S. Shrestha;Michael Hahn - 通讯作者:
Michael Hahn
Michael Hahn的其他文献
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{{ truncateString('Michael Hahn', 18)}}的其他基金
Collaborative Research: SHINE: Observational and Theoretical Studies of the Parametric Decay Instability in the Lower Solar Atmosphere
合作研究:SHINE:太阳低层大气参数衰变不稳定性的观测和理论研究
- 批准号:
2229100 - 财政年份:2023
- 资助金额:
$ 54.7万 - 项目类别:
Standard Grant
High-Resolution Observations of Alfvenic Waves in the Solar Corona: Critical Early DKIST Science
日冕中阿尔芬波的高分辨率观测:关键的早期 DKIST 科学
- 批准号:
2005887 - 财政年份:2020
- 资助金额:
$ 54.7万 - 项目类别:
Standard Grant
Understanding Wave Energy Transport Through the Complex Chromosphere and Transition Region
了解复杂色球层和过渡区域的波能传输
- 批准号:
1834822 - 财政年份:2019
- 资助金额:
$ 54.7万 - 项目类别:
Standard Grant
SHINE: Observational Constraints on Wave Heating of the Corona
SHINE:日冕波加热的观测限制
- 批准号:
1459247 - 财政年份:2015
- 资助金额:
$ 54.7万 - 项目类别:
Continuing Grant
A Toolkit for in Vivo Visualization/Modulation of Plant Cell Wall Polysaccharides
植物细胞壁多糖体内可视化/调节的工具包
- 批准号:
0923992 - 财政年份:2010
- 资助金额:
$ 54.7万 - 项目类别:
Continuing Grant
A Monoclonal Antibody Toolkit for Functional Genomics of Plant Cell Walls
用于植物细胞壁功能基因组学的单克隆抗体工具包
- 批准号:
0421683 - 财政年份:2004
- 资助金额:
$ 54.7万 - 项目类别:
Continuing Grant
Purification and Cloning of Hepta-beta Glucoside Elicitor- binding Protein(s) from Soybean
大豆中七-β葡萄糖苷激发子结合蛋白的纯化和克隆
- 批准号:
9723685 - 财政年份:1997
- 资助金额:
$ 54.7万 - 项目类别:
Continuing Grant
Isolation of a Receptor for a Fungal Wall Derived Elicitor of Phytoalexins
真菌壁衍生的植物抗毒素激发子受体的分离
- 批准号:
8904574 - 财政年份:1989
- 资助金额:
$ 54.7万 - 项目类别:
Continuing Grant
Isolation of a Receptor for a Fungal-Wall-Derived Eliitor of Phytoalexins
真菌壁衍生的植物抗毒素Eliitor受体的分离
- 批准号:
8704022 - 财政年份:1987
- 资助金额:
$ 54.7万 - 项目类别:
Standard Grant
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