基于荧光素的电泳迁移率变动分析。

Fluorescein-Based Electrophoretic Mobility Shift Assay.

机构信息

Department of Biological Sciences, Mississippi State University, Mississippi State, MS, USA.

出版信息

Methods Mol Biol. 2022;2316:133-140. doi: 10.1007/978-1-0716-1464-8_12.

Abstract

RNA-protein complexes are functionally important in biology. Electrophoretic mobility shift assays (EMSA) have been widely used to study the molecular basis of protein-RNA interactions. Previous methods for EMSA mostly relied on radioactive RNA substrates, raising health and environmental concerns. Here, we describe a method based on fluorescein-labeled RNA for EMSA. In addition, we simplified the protocol to efficiently purify RNA-binding proteins from bacterial expression systems.

摘要

RNA-蛋白质复合物在生物学中具有重要的功能。电泳迁移率变动分析（EMSA）已被广泛用于研究蛋白质-RNA 相互作用的分子基础。之前的 EMSA 方法大多依赖于放射性 RNA 底物，这引起了健康和环境方面的关注。在这里，我们描述了一种基于荧光素标记 RNA 的 EMSA 方法。此外，我们简化了从细菌表达系统中有效纯化 RNA 结合蛋白的方案。