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甾醇产生分枝杆菌的细胞内和细胞外蛋白质组的二维分析。

Bidimensional Analyses of the Intra- and Extracellular Proteomes of Steroid Producer Mycobacteria.

机构信息

Área de Bioquímica y Biología Molecular, Departamento de Biología Molecular, Facultad de Veterinaria, Universidad de León, León, Spain.

Instituto de Investigación de la Viña y el Vino, Escuela de Ingeniería Agraria, Universidad de León, León, Spain.

出版信息

Methods Mol Biol. 2023;2704:115-141. doi: 10.1007/978-1-0716-3385-4_7.

Abstract

The importance of the pathogenic mycobacteria has mainly focused the omic analyses on different aspects of their clinical significance. However, those industrially relevant mycobacteria have received less attention, even though the steroid market sales in 2021 were estimated in $56.45 billion.The extracellular proteome, due to its relevance in the sterol processing and uptake, and the intracellular proteome, because of its role in steroids bioconversion, are the core of the present chapter. Both, monodimensional gels, as preparatory analysis, and bidimensional gels as proteome analysis are described. As a proof of concept, the protein extraction methods for both sub-proteomes of Mycobacterium are described. Thus, procedures and relevant key points of these proteome analyses are fully detailed.

摘要

致病分枝杆菌的重要性主要集中在它们临床意义的不同方面的组学分析上。然而,那些与工业相关的分枝杆菌受到的关注较少,尽管 2021 年类固醇市场销售额估计为 564.5 亿美元。细胞外蛋白质组由于其在甾醇加工和摄取中的相关性,以及细胞内蛋白质组由于其在类固醇生物转化中的作用,是本章的核心。本文描述了一维凝胶作为预备分析和二维凝胶作为蛋白质组分析。作为概念验证,还描述了分枝杆菌这两个亚蛋白质组的蛋白质提取方法。因此,详细说明了这两种蛋白质组分析的程序和相关要点。

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