Université de Lorraine, CNRS, IMoPA, F-54000 Nancy, France.
Methods Mol Biol. 2021;2300:99-106. doi: 10.1007/978-1-0716-1386-3_10.
RiboNucleoProtein particles (RNPs), which are composed of RNAs and proteins, play essential roles in many biological processes. The isolation of these molecular machines is a critical step to better understand their mechanisms of action. In this chapter, we describe the MS2-MBP affinity chromatography used to purify the protein content of the RNPs formed with an RNA of interest in a nuclear extract. Substrate RNAs are furnished with a tag consisting of three stem-loops that provide specific binding sites for the phage MS2 protein. Here, we successfully applied this method to isolate RNPs formed with subfragments of the long noncoding RNA ANRIL (Antisense Noncoding RNA in the INK4 Locus).
核糖核蛋白颗粒（RNPs）由 RNA 和蛋白质组成，在许多生物过程中发挥着重要作用。这些分子机器的分离是更好地了解其作用机制的关键步骤。在本章中，我们描述了 MS2-MBP 亲和层析法，用于纯化用感兴趣的 RNA 在核提取物中形成的 RNPs 的蛋白质含量。底物 RNA 带有一个标记，该标记由三个茎环组成，为噬菌体 MS2 蛋白提供了特定的结合位点。在这里，我们成功地将该方法应用于分离长非编码 RNA ANRIL（INK4 基因座反义非编码 RNA）的亚片段形成的 RNPs。
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