Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Methods. 2020 Jun 1;178:83-88. doi: 10.1016/j.ymeth.2019.09.001. Epub 2019 Sep 5.
mRNA binding proteins (RBPs) play a major role in post-transcriptional control of gene expression. To understand the complex regulatory processes regulating a specific mRNA during its life-time, a comprehensive view of the bound RBPs is essential. Here, we describe a method for transcript-specific isolation of endogenous ribonucleoprotein complexes (RNPs) from Drosophila egg-chambers. The method, which is based on in-solution hybridization of short biotinylated antisense DNA oligonucleotide probes to multiple segments of a transcript of interest allows unbiased identification of associated proteins by quantitative proteomics.
mRNA 结合蛋白（RBPs）在后转录基因表达调控中发挥着重要作用。为了了解在特定 mRNA 的整个生命周期中调节其的复杂调控过程，全面了解结合的 RBPs 是至关重要的。在这里，我们描述了一种从果蝇卵室中分离内源性核糖核蛋白复合物（RNP）的方法，该方法基于短的生物素化反义 DNA 寡核苷酸探针与感兴趣的转录本的多个片段在溶液中的杂交，通过定量蛋白质组学可无偏地鉴定相关蛋白。
