Arabidopsis 2010 Project: Development of Arabidopsis Proteome Chip

拟南芥的全基因组序列揭示了大量的新基因，但DNA序列本身对其具体功能提供的线索很少。目前，人们对基因的功能和表达进行了大量的研究。虽然这些研究很重要，但还不足以预测细胞中蛋白质的结构、功能和活性。蛋白质组学是一种对蛋白质进行全局分析的方法，它正在成为一个重要的研究领域。在这个试点项目中，将生成一个小的拟南芥开放阅读框表达克隆集合。利用这些收集，将开发最佳的蛋白表达系统来生产和纯化拟南芥蛋白。这些蛋白质将被打印以产生先导蛋白质芯片，然后将被筛选用于各种生化分析，包括蛋白质-蛋白质相互作用分析，蛋白质- dna /RNA相互作用，蛋白质-磷脂相互作用以及激酶和其他酶的底物鉴定。更广泛的影响：大规模生物学领域的快速发展为理解生物网络的整体功能提供了机会。由于基因的生化功能是通过其编码的蛋白质来体现的，因此近年来蛋白质组学的新工具的开发受到了很大的重视。因此，该项目最终将为科学界提供巨大的宝贵资源，用于旨在高通量研究拟南芥蛋白质功能的各种应用。蛋白质组芯片的优势在于，一套全面的单个蛋白质可以直接在体外筛选各种各样的活性。此外，一旦蛋白质准备好，蛋白质组筛选比其他方法要快得多，也便宜得多。该项目产生的试剂和信息将定期提供给科学界（http://bioinfo.mbb.yale.edu/genome/plant），预计将大大加强对拟南芥蛋白质功能的分析。

The complete genome sequence of Arabidopsis has revealed a large number of novel genes, however DNA sequence alone offers few clues as to their specific functions. Currently, much effort is being devoted toward studying gene function and expression. Although important, these studies are not sufficient to predict the structure, function, and activity of proteins in the cell. Proteomics, the global analysis of proteins, is emerging as an important area of research. In this pilot project, a small collection of expression clones of Arabidopsis open reading frames will be generated. Using this collection, optimal protein expression systems will be developed to produce and purify Arabidopsis proteins. The proteins will be printed to generate pilot protein chips, which will then be screened for various biochemical assays, including protein-protein interaction assays, protein-DNA/RNA interactions, protein-phospholipids interactions and the identification of substrates for kinases and other enzymes.Broader Impacts: The rapid progress in the field of large-scale biology has provided the opportunity to understand the function of biological networks as a whole. Since the biochemical function of a gene is manifested through its encoded protein, recently much emphasis has been devoted to the development of new tools for proteomics. Therefore, this project will ultimately provide an enormous valuable resource to the scientific community for a variety of applications aimed at the high-throughput study of protein function in Arabidopsis. The advantage of proteome chips is that a comprehensive set of individual proteins can be directly screened in vitro for a wide variety of activities. Furthermore, once the proteins are prepared, proteome screening is significantly faster and cheaper than other methods. The reagents and information generated from this project will be available to the scientific community (http://bioinfo.mbb.yale.edu/genome/plant) on a regular basis and are expected to significantly enhance the analysis of protein function in Arabidopsis.