Arabidopsis 2010: Development of an Arabidopsis proteome chip

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线索的反应中是如何发挥作用的。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，本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，优化了用于克隆和表达1000个拟南芥ORF的高通量技术，以产生用于产生拟南芥蛋白芯片的高质量活性蛋白。在这个项目中，4,000个预测的拟南芥ORF的表达克隆的集合将产生串联亲和纯化（TAP）标签融合。这些蛋白将在植物瞬时表达系统中表达，以生产和纯化蛋白。这些蛋白质将被打印在各种打印表面上，以产生蛋白质微阵列，然后将用于优化蛋白质活性分析的方案。产生的试剂将提供给整个科学界，信息将通过我们的网站（http：//www.gersteinlab.org/proj/atpchip/）。更广泛的影响：该项目将提供一个有价值的资源，用于各种应用，旨在高通量研究蛋白质功能的拟南芥。该项目将产生一套特定于植物蛋白质组的方案，并产生一组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，从这项研究中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。除了产生一个社区资源，该项目提供了独特的机会，以提高认识和基因组学和蛋白质组学的重要性，以广泛的个人，包括来自世界各地的访问学者，教育工作者从小型学院，公立中学教师和周围的纽黑文地区和代表性不足的学生在本科和研究生水平。具体来说，参与耶鲁大学的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尖端研究的机会。

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.