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功能蛋白质组学分析方法。

Methods for functional proteomic analyses.

作者信息

Rampitsch Christof, Bykova Natalia V

机构信息

Agriculture and Agri-food Canada, Cereal Research Centre, Winnipeg, MB R3T 2M9, Canada.

出版信息

Methods Mol Biol. 2009;513:93-110. doi: 10.1007/978-1-59745-427-8_6.

Abstract

The term 'Proteomics' was introduced in 1997 to describe a growing interest in the study of the proteome - the expressed protein set of an organism. As this new discipline evolved, it quickly became obvious that proteomics would be a very complex and ambitious undertaking, perhaps even more so than genomics, which had engendered it. New techniques for both the separation and analysis/identification of proteins were emerging or being refined, and these facilitated the development of this new field. Many proteomics experiments are now routine in some laboratories. In this chapter we describe a typical proteomics experiment, using examples from our laboratory: the separation of complex mixtures of proteins by 2-dimensional electrophoresis and subsequent identification of a protein spot by mass spectrometry with two commonly used instruments: MALDI-QqTOF and ESI-ion trap.

摘要

“蛋白质组学”一词于1997年被提出,用于描述对蛋白质组(生物体中表达的蛋白质集合)研究日益增长的兴趣。随着这一新学科的发展,很快就变得明显的是,蛋白质组学将是一项非常复杂且雄心勃勃的任务,可能甚至比催生它的基因组学更为复杂。用于蛋白质分离以及分析/鉴定的新技术不断涌现或得到改进,这些都推动了这一新领域的发展。现在,许多蛋白质组学实验在一些实验室中已成为常规操作。在本章中,我们将以我们实验室的实例描述一个典型的蛋白质组学实验:通过二维电泳分离复杂的蛋白质混合物,随后使用两种常用仪器——基质辅助激光解吸电离串联四极杆飞行时间质谱仪(MALDI-QqTOF)和电喷雾离子阱质谱仪(ESI-ion trap),通过质谱法鉴定一个蛋白质斑点。

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