Mitchell Sarah F, Parker Roy
Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado Boulder, Jennie Smoly Caruthers Biotech Bldg., 3415 Colorado Avenue, Boulder, CO, 80303, USA.
Methods Mol Biol. 2016;1421:153-63. doi: 10.1007/978-1-4939-3591-8_13.
The maturation, localization, stability, and translation of messenger RNAs (mRNAs) are regulated by a wide variety of mRNA-binding proteins. Identification of the complete set of mRNA-binding proteins is a key step in understanding the regulation of gene expression. Herein, we describe a method for identifying yeast mRNA-binding proteins in a systematic manner using UV crosslinking, purification of polyA(+) mRNAs under denaturing conditions, and mass spectrometry to identify covalently bound proteins.
信使核糖核酸（mRNA）的成熟、定位、稳定性及翻译受多种mRNA结合蛋白调控。鉴定出完整的mRNA结合蛋白集是理解基因表达调控的关键一步。在此，我们描述了一种系统鉴定酵母mRNA结合蛋白的方法，该方法利用紫外线交联、在变性条件下纯化聚腺苷酸（polyA(+)）mRNA以及通过质谱鉴定共价结合蛋白。
