Song Guang, Neiswinger Johnathan, Zhu Heng
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287.
Cold Spring Harb Protoc. 2016 Oct 3;2016(10):2016/10/pdb.prot087973. doi: 10.1101/pdb.prot087973.
RNA-binding proteins (RBPs), along with target RNA, play a vital role in the regulation of cellular processing and are especially important in gene transcription and posttranscriptional regulation. Here, we present a high-throughput method for rapid identification of RBPs for a given RNA using protein microarray technology. This protocol includes preparation of Cy5-labeled RNA probes, probe denaturing and refolding, and an RNA-binding assay as performed on a yeast protein microarray.
RNA结合蛋白（RBPs）与靶RNA一起，在细胞加工过程的调控中起着至关重要的作用，在基因转录和转录后调控中尤为重要。在这里，我们介绍一种使用蛋白质微阵列技术快速鉴定给定RNA的RBPs的高通量方法。该方案包括Cy5标记的RNA探针的制备、探针变性和复性，以及在酵母蛋白质微阵列上进行的RNA结合测定。
