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一种基于十六烷基三甲基溴化铵的方法,用于制备用于蛋白质组学分析的葡萄酒腐败微生物总蛋白提取物。

A CTAB based method for the preparation of total protein extract of wine spoilage microrganisms for proteomic analysis.

作者信息

Polati Rita, Zapparoli Giacomo, Giudici Paolo, Bossi Alessandra

机构信息

Dipartimento Scientifico e Tecnologico, Università di Verona, 37134 Verona, Italy.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Apr 1;877(10):887-91. doi: 10.1016/j.jchromb.2009.02.022. Epub 2009 Feb 11.

Abstract

Mapping the proteome of microrganisms by 2D-electrophoresis is often a hard task, because many contaminants, e.g. polysaccharides of the cell wall and nucleic acid, can obstruct the pores of the IEF gel resulting in streaks and smears. A protocol based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) and its salt-dependent solubility was developed. The cellulose-producing strain Gluconoacetobacter hansenii AAB0248 was resolved on 7cm Minigels in over 500 protein spots (a hundred more than with protocols reported in literature). The method was further employed for mapping the proteome of some acid adapted, wine spoilage microrganisms e.g. acetic acid bacteria and a yeast.

摘要

通过二维电泳对微生物蛋白质组进行图谱分析往往是一项艰巨的任务,因为许多污染物,如细胞壁多糖和核酸,会堵塞等电聚焦凝胶的孔隙,导致条带和拖尾现象。基于使用阳离子去污剂十六烷基三甲基溴化铵(CTAB)及其盐依赖性溶解度,开发了一种方法。产纤维素菌株汉逊醋杆菌AAB0248在7厘米的微型凝胶上分离出500多个蛋白质点(比文献报道的方法多一百多个)。该方法进一步用于绘制一些适应酸性环境的葡萄酒腐败微生物(如醋酸菌和一种酵母)的蛋白质组图谱。

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