Peptide aptamers to investigate and disrupt protein function in plants

用于研究和破坏植物蛋白质功能的肽适体

• 批准号: 0926350
• 负责人: Stanton Gelvin
• 金额: $ 30万
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0926350&HistoricalAwards=false
• 关键词: Peptide; aptamers; investigate; disrupt; protein

PI: Stanton B. Gelvin (Purdue University)CoPI: Vitaly Citovsky (State University of New York, Stony Brook) Aptamers are small synthetic peptides of random amino acid sequence. When expressed in cells, aptamers can specifically bind to target proteins and inhibit function. They can thus be used to define the functional roles of proteins in various cellular processes, as well as defining specific domains within these proteins that are important for the proteins to mediate these roles. Furthermore, the aptamer sequences can be used as functional identification "marks" of plant proteins that naturally interact with the target proteins. An expression vector and libraries of millions of random aptamers will be developed for use in transient transformation experiments. The vector will allow expression and screening of aptamers first in yeast, and subsequently in plants. The constructions will incorporate bimolecular fluorescence complementation (BiFC) technology to detect aptamer-protein interactions, and Gateway technologies to facilitate mobilization of aptamers among various vectors. As proof of concept, tagged plant proteins important for development, and Agrobacterium virulence proteins important for Agrobacterium-mediated transformation will be probed with the aptamer library. This initial work will test whether random aptamers can be used to identify interacting proteins in two different organisms, yeast and plants. This project will provide research tools and resources for the plant science community to determine the functions of plant proteins, and protein-protein interactions in planta. Data will be released sequentially through a project website at Purdue University (accessible via http://bio.purdue.edu/people/faculty/index.php?refID=128) and the aptamer library will be provided to the ABRC for distribution.

PI：Stanton B. Gelvin（Purdue University）CoPI：Vitaly Citovsky（州立大学，纽约，斯托尼布鲁克）适体是随机氨基酸序列的小的合成肽。 当在细胞中表达时，适体可以特异性地结合靶蛋白并抑制功能。 因此，它们可以用于定义蛋白质在各种细胞过程中的功能作用，以及定义这些蛋白质中对蛋白质介导这些作用很重要的特定结构域。 此外，适体序列可用作与靶蛋白天然相互作用的植物蛋白的功能鉴定"标记"。 将开发表达载体和数百万个随机适体的文库，用于瞬时转化实验。 该载体将允许首先在酵母中，随后在植物中表达和筛选适体。 这些构建将结合双分子荧光互补（BiFC）技术来检测适体与蛋白质的相互作用，以及Gateway技术来促进适体在各种载体之间的动员。 作为概念的证明，将用适体文库探测对发育重要的标记的植物蛋白和对农杆菌介导的转化重要的农杆菌毒力蛋白。 这项初步工作将测试随机适体是否可以用于识别两种不同生物体（酵母和植物）中相互作用的蛋白质。该项目将为植物科学界提供研究工具和资源，以确定植物蛋白质的功能以及植物中蛋白质-蛋白质相互作用。数据将通过普渡大学的项目网站（可通过www.example.com访问）按顺序发布refID = 128），并将适体文库提供给ABRC用于分发。