Fudan University Zhongshan Hospital, Shanghai, P. R. China.
Methods Mol Biol. 2021;2372:27-33. doi: 10.1007/978-1-0716-1697-0_4.
Long non-coding RNAs (lncRNAs) are a new class of regulatory genes that play critical roles in various processes ranging from normal development to human diseases. Recent studies have shown that protein complexes are required for the functions of lncRNAs. The identification of these proteins which are associated with lncRNAs is critical for the understanding of molecular mechanisms of lncRNAs in gene regulation and their functions. In this chapter, we describe a method to isolate proteins associated with lncRNAs. This procedure involves fusion protein Maltose-Binding Protein (MBP) fused to MS2-binding protein to pull down the proteins associated with lncRNA and the identification of these proteins by mass spectrometry.
长非编码 RNA(lncRNAs)是一类新的调控基因,在从正常发育到人类疾病等各种过程中发挥着关键作用。最近的研究表明,蛋白质复合物是 lncRNA 功能所必需的。鉴定与 lncRNA 相关的这些蛋白质对于理解 lncRNA 在基因调控及其功能中的分子机制至关重要。在本章中,我们描述了一种分离与 lncRNA 相关的蛋白质的方法。该程序涉及融合蛋白麦芽糖结合蛋白(MBP)与 MS2 结合蛋白融合,以拉下与 lncRNA 相关的蛋白质,并通过质谱鉴定这些蛋白质。