Arabidopsis 2010: Development of an Arabidopsis proteome chip

拟南芥 2010：拟南芥蛋白质组芯片的开发

• 批准号: 0519853
• 负责人: Michael Snyder
• 金额: $ 150万
• 依托单位: Yale University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-11-01 至 2008-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0519853&HistoricalAwards=false
• 关键词: Arabidopsis; 2010; Development; proteome; chip

The genome sequence of Arabidopsis suggests that it contains approximately 30,700 protein-coding genes. A major goal of the 2010 program is to understand how each of these genes function during plant growth, development, and responses to biotic and abiotic cues. Global studies to analyze gene and protein function have largely focused on gene expression, gene disruption, protein interactions and protein localization. To elucidate biochemical activities of the proteome at the global scale, this project will continue to develop a protein chip for Arabidopsis. In the pilot project funded by NSF 2010 program, high throughput techniques for cloning and expression of 1000 Arabidopsis ORFs were optimized to produce high quality active proteins for the generation of an Arabidopsis protein chip. In this project, a collection of expression clones for 4,000 predicted Arabidopsis ORFs will be generated for tandem affinity purification (TAP) tag fusions. These proteins will be expressed in the plant-based transient expression system to produce and purify proteins. The proteins will be printed on various printing surfaces to produce protein microarrays, which will then be used to optimize protocols for analysis of protein activities. The reagents generated will be made available to the entire scientific community and the information will be available through our website (http://www.gersteinlab.org/proj/atpchip/).Broader Impacts: This project will provide a valuable resource for a variety of applications aimed at the high-throughput study of protein function in Arabidopsis. The project will generate a suite of protocols specific to plant proteomes and generate a resourceful set of plant expression clones and other reagents that are expected to significantly enhance the analysis of protein function in Arabidopsis. Furthermore, the methods developed and information gained from this study can also be directly applied to the analysis of other agriculturally and horticultural plants. In addition to generating a community resource, the project provides the unique opportunity to elevate the awareness and importance of genomics and proteomics to a wide range of individuals including visiting scholars from around the world, educators from small colleges, public secondary school teachers in and around the New Haven area and under represented students at the undergraduate and graduate level. Specifically, participation in Yale University STARS program will provide opportunities for under represented students to conduct cutting edge research in proteomics lab.

拟南芥的基因组序列表明它包含大约 30,700 个蛋白质编码基因。 2010 年计划的一个主要目标是了解这些基因在植物生长、发育以及对生物和非生物信号的反应过程中如何发挥作用。分析基因和蛋白质功能的全球研究主要集中在基因表达、基因破坏、蛋白质相互作用和蛋白质定位。为了在全球范围内阐明蛋白质组的生化活性，该项目将继续开发拟南芥蛋白质芯片。在 NSF 2010 计划资助的试点项目中，对 1000 个拟南芥 ORF 的克隆和表达的高通量技术进行了优化，以产生用于生成拟南芥蛋白质芯片的高质量活性蛋白质。在该项目中，将生成 4,000 个预测拟南芥 ORF 的表达克隆集合，用于串联亲和纯化 (TAP) 标签融合。这些蛋白质将在基于植物的瞬时表达系统中表达以产生和纯化蛋白质。这些蛋白质将被印刷在各种印刷表面上以产生蛋白质微阵列，然后将其用于优化蛋白质活性分析的方案。生成的试剂将提供给整个科学界，并且信息将通过我们的网站 (http://www.gersteinlab.org/proj/atpchip/) 提供。 更广泛的影响：该项目将为旨在拟南芥蛋白质功能高通量研究的各种应用。该项目将生成一套针对植物蛋白质组的实验方案，并生成一套资源丰富的植物表达克隆和其他试剂，预计将显着增强拟南芥蛋白质功能的分析。此外，本研究开发的方法和获得的信息也可以直接应用于其他农业和园艺植物的分析。除了产生社区资源外，该项目还提供了独特的机会，以提高广大个人对基因组学和蛋白质组学的认识和重要性，包括来自世界各地的访问学者、小型学院的教育工作者、国内外公立中学教师纽黑文地区本科生和研究生的学生人数较少。具体来说，参与耶鲁大学 STARS 计划将为代表性不足的学生提供在蛋白质组学实验室进行前沿研究的机会。