Rice proteome analysis as a post-genome project research

为了有效地分析水稻蛋白质组，我们在本研究中发展了各种分析技术。这些技术包括蛋白质凝胶内消化技术、肽图谱技术、双向电泳法分离蛋白质的质谱学技术、十二烷基硫酸钠凝胶电泳法分离蛋白质的枪法分析、磷酸化蛋白质的检测、高相对分子质量蛋白质的电泳法，以及新型蛋白质芯片的蛋白质-蛋白质相互作用分析。利用所开发的技术，我们对水稻种子胚胎蛋白质组进行了全面的分析。在这项分析中，我们鉴定了许多未知功能的蛋白质。为了获得这些蛋白质的功能信息，我们对它们进行了表达谱分析，并鉴定了几个与萌发相关的蛋白质。另一方面，我们分析了一个大型蛋白质复合体26S蛋白酶体，以确定蛋白质翻译后修饰的作用，为蛋白质组分析提供基础信息。在我们的蛋白质组数据库中编辑了单个胚胎蛋白质的双向电泳谱和特征。

To analyze efficiently the rice proteome, we developed various analytical techniques in the present study. These include techniques for protein in-gel digestion, peptide mapping, mass spectrometry for identification of proteins separated by two-dimensional electrophoresis, shot-gun analysis of proteins separated by SDS gel electrophoresis, detection for phosphorylated proteins, electrophoresis for high molecular weight proteins, and protein-protein interaction analysis with a novel protein chip. Using the developed techniques, we analyzed comprehensively the rice seed embryo proteome. In this analysis, we identified many proteins without known functions. To obtain the information of functions of these proteins, we performed expression profiling of them, and we identified several proteins, which expressed in relation to germination. On the other hand, we analyzed a large protein complex, 26S proteasome to determine the roles of protein post-translational modifications to obtain fundamental information for proteome analysis. The two-dimensional electrophoresis pattern and the characteristics of individual embryo protein were compiled in our proteome database.