Electrophoretic mobility shift assays (EMSAs) for in vitro detection of protein-nucleic acid interactions

电泳迁移率变动分析 (EMSA) 用于体外检测蛋白质-核酸相互作用

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作者:Farnaz Mansouri-Noori ,Mark A Bayfield

Abstract

Protein-nucleic acid interactions drive some of the most important physiological events in cells. Here, we present a protocol for detecting protein-DNA or protein-RNA interactions in vitro. We describe steps for labeling nucleic acid species and electrophoretic mobility shift assays (EMSAs). This protocol can be used to confirm suspected in vivo interactions using recombinantly expressed/purified proteins of interest and a nucleic acid substrate. It can further be used to investigate mutations that can disrupt interaction or compensatory mutations that restore it. For complete details on the use and execution of this protocol, please refer to Mansouri-Noori et al.1. Keywords: Gene Expression; Molecular Biology; Molecular/Chemical Probes; Protein Biochemistry.

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