ANALYSIS OF THE YEAST PROTEOME USING PROTEIN CHIPS

描述(逐字摘自申请者摘要)：关于MOST的信息 酿酒酵母基因，无论是新发现的还是以前发现的 辨认出来的，是非常有限的。目前，对基因的研究越来越受到重视。 在核酸水平上的分析，如对mRNA基因的研究 表达或基因组变异。然而，基因的功能通常是在 蛋白质分析水平，总体上没有受到很高的影响 吞吐量技术。我们建议开发高吞吐量的方法和 用于蛋白质功能的大规模生化分析的蛋白质芯片 酵母。将准备一组酵母菌株，将酵母ORF表达为GST 在半乳糖诱导启动子的控制下融合；我们将 随后使用高通量方法进行大规模表达 酵母蛋白。这些蛋白质将被附着在新型微孔芯片上， 正在进行生化活动的开发和分析。具体来说，我们将 使用体外结合和/或酶试验鉴定GTP结合的靶标 蛋白质、磷脂结合蛋白和蛋白磷酸酶。我们还将 利用蛋白质进一步分析蛋白激酶及其体外底物 微芯片技术。 我们预计这个项目将对科学研究产生重大影响 社区。生成的信息将在一个数据库中维护，该数据库 可通过互联网访问。我们的质粒和酵母菌株将会被制造出来 可供所有感兴趣的研究人员使用。最重要的是技术 预计将对酵母和酵母的研究产生巨大影响 其他生物。

DESCRIPTION (Verbatim from Applicant's Abstract): Information about most Saccharomyces cerevisiae genes, whether newly discovered or previously identified, is very limited. Currently, much emphasis has been placed on gene analysis at the level of nucleic acids such as the study of mRNA gene expression or genome variation. However, gene function is often deduced at the level of protein analysis, which in general has not been subjected to high throughput techniques. We propose to develop high throughput methods and protein chips for the large-scale biochemical analysis of protein function in yeast. A set of yeast strains will be prepared that express yeast ORFs as GST fusions under the control of a galactose-inducible promoter; we will subsequently use high throughput methods for the large-scale expression of the yeast proteins. The proteins will be adhered to novel microwell chips that we are developing and analyzed for biochemical activities. Specifically, we will use in vitro binding and/or enzymatic assays to identify targets of GTP-binding proteins, phospholipid binding protein and protein phosphatases. We will also further analyze protein kinases and their in vitro substrates using protein microchip techniques. We expect this project to have a significant impact on the scientific community. The information generated will be maintained in a database that is accessible over the Internet. Our plasmids and yeast strains will be made available to all interested researchers. Most importantly the technology developed is expected to have an enormous impact on the study of yeast and other organisms.