Department of Microbial Sciences, School of Biosciences and Medicine, Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey GU2 7XH, UK.
STAR Protoc. 2021 Nov 1;2(4):100929. doi: 10.1016/j.xpro.2021.100929. eCollection 2021 Dec 17.
characterization of RNA-protein interactions is the key for understanding RNA regulatory mechanisms. Herein, we describe a protocol for detection of proteins interacting with polyadenylated RNAs in the yeast Proteins are crosslinked to nucleic acids by ultraviolet (UV) irradiation of cells, and poly(A)-containing RNAs with bound proteins are isolated from cell lysates using oligo[dT] beads. RBPs can be detected by immunoblot analysis or with mass spectrometry to define the mRNA-binding proteome (mRBPome) and its changes under stress. For complete details on the use and execution of this protocol, please refer to Matia-González et al. (2021, 2015).
RNA-蛋白质相互作用的表征是理解 RNA 调控机制的关键。在此，我们描述了一种在酵母中检测与多聚腺苷酸化 RNA 相互作用的蛋白质的方案。通过对细胞进行紫外线（UV）照射，将蛋白质交联到核酸上，然后使用 oligo[dT]珠从细胞裂解物中分离出与结合蛋白结合的多聚（A）RNA。通过免疫印迹分析或质谱分析可以检测 RBPs，以定义应激下的 mRNA 结合蛋白质组（mRBPome）及其变化。有关此方案的使用和执行的完整详细信息，请参阅 Matia-González 等人（2021 年，2015 年）。
Zotero 插件
