Klose J
Institut für Humangenetik, Freie Universität Berlin, Federal Republic of Germany.
Electrophoresis. 1989 Feb;10(2):140-52. doi: 10.1002/elps.1150100208.
High-resolution two-dimensional electrophoresis (2-DE) has reached a technological level that allows us to resolve most of the numerous unknown protein species of a mammalian organism if appropriate strategies are used. We will discuss the problems of classification and characterization of proteins and propose a systematic approach to the analysis of the total protein complex. Both a comprehensive as well as a pragmatic approach towards systematic analysis have been considered. A "complex protein database" is suggested and considered with regard to various uses. A systematic analysis of the mouse proteins has been started and some of the preliminary results are summarized here. In particular, genetic properties of the proteins were investigated and are presented in order to demonstrate the significance of a systematic analysis of proteins for research and practical application (e.g. mutagenicity testing). A concept is presented for sequencing the coding DNA of mouse and man, starting with a systematic analysis of mouse proteins and then using two recently developed methods - microsequencing of proteins from spots of 2-DE protein patterns, and utilization of the relatively short N-terminal sequences obtained - to produce the corresponding cDNA's of these proteins.
高分辨率二维电泳（2-DE）已经达到了这样一个技术水平：如果采用适当的策略，我们就能解析哺乳动物机体中大多数数量众多的未知蛋白质种类。我们将讨论蛋白质的分类和表征问题，并提出一种分析总蛋白复合物的系统方法。我们考虑了对系统分析采用全面和务实的方法。提出并考虑了一个“复杂蛋白质数据库”的各种用途。已经开始对小鼠蛋白质进行系统分析，这里总结了一些初步结果。特别是，对蛋白质的遗传特性进行了研究并呈现出来，以证明蛋白质系统分析对于研究和实际应用（如致突变性测试）的重要性。本文提出了一个对小鼠和人类编码DNA进行测序的概念，首先对小鼠蛋白质进行系统分析，然后使用两种最近开发的方法——对2-DE蛋白质图谱斑点中的蛋白质进行微量测序，以及利用获得的相对较短的N端序列——来产生这些蛋白质的相应cDNA。
