Affinity purification of ribosomes to access the translatome
Affinity purification of ribosomes to access the translatome
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DOI：
10.1016/j.ymeth.2009.04.003
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发表时间：
2009-07-01
期刊：
影响因子：
4.8
通讯作者：
Gerber, Andre P.
中科院分区：
文献类型：
--
作者：
Halbeisen, Regula E.;Scherrer, Tanja;Gerber, Andre P.
我们描述了核糖体亲和纯化（RAP），这是一种可以快速纯化来自酿酒酵母的核糖体和相关信息的方法。该方法依赖于核糖体蛋白 Rpl16 的蛋白 A 标记版本的表达，该蛋白用于通过 IgG 偶联的球形微珠从细胞提取物中有效回收内源形成的核糖体和多聚核糖体。这种方法可以应用于分析翻译组的反应，翻译组是指与核糖体相关的所有信息，以及使用 DNA 微阵列的转录组的反应。此外，核糖体蛋白、它们的修饰和/或其他相关蛋白可以通过质谱法进行定位。最后，这种方法在其他生物体中的应用为破译细胞类型特异性基因表达模式提供了有价值的工具。 (C) 2009 Elsevier Inc. 保留所有权利。
We describe ribosome affinity purification (RAP), a method that allows rapid purification of ribosomes and associated messages from the yeast Saccharomyces cerevisiae. The method relies on the expression of protein A tagged versions of the ribosomal protein Rpl16, which is used to efficiently recover endogenously formed ribosomes and polysomes from cellular extracts with IgG-coupled spherical microbeads. This approach can be applied to profile reactions of the translatome, which refers to all messages associated with ribosomes, with those of the transcriptome using DNA microarrays. In addition, ribosomal proteins, their modifications, and/or other associated proteins can be mapped with mass spectrometry. Finally, application of this method in other organisms provides a valuable tool to decipher cell-type specific gene expression patterns. (C) 2009 Elsevier Inc. All rights reserved.