Liu Song, Liu Bing, Zheng Chunfu
School of Pharmacy & Clinical Pharmacy, School of Integrative Pharmacy, Guangdong Pharmaceutical University, Guangzhou, China.
Department of Microbiology, Immunology & Infectious Diseases, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2025;2940:343-355. doi: 10.1007/978-1-0716-4615-1_30.
Many viral proteins bind to DNA and play critical roles in the protection or stabilization of the viral genome during infection, capsid shell synthesis, and entire lifespan of the virus. Moreover, the interaction between a viral protein or host cell protein and its DNA can regulate virus recognition, host infection, and host-virus interactions and affect antiviral immunity. Thus, a method is needed to confirm the interaction between viral proteins and DNA, which would provide a better understanding of the mechanism and function of viral proteins and DNA and the regulation of antiviral immunity. An electrophoretic mobility shift assay (EMSA) with a biotin-labeled probe provides a simple way to determine the interaction between viral proteins and DNA during the virus lifecycle and antiviral immunity. This chapter presents detailed information and experimental procedures for measuring the interaction between target proteins and DNA via EMSA.
许多病毒蛋白会与DNA结合,并在感染、衣壳合成以及病毒的整个生命周期中对病毒基因组的保护或稳定发挥关键作用。此外,病毒蛋白或宿主细胞蛋白与其DNA之间的相互作用可调节病毒识别、宿主感染以及宿主-病毒相互作用,并影响抗病毒免疫。因此,需要一种方法来确认病毒蛋白与DNA之间的相互作用,这将有助于更好地理解病毒蛋白和DNA的机制与功能以及抗病毒免疫的调节。使用生物素标记探针的电泳迁移率变动分析(EMSA)为确定病毒生命周期和抗病毒免疫过程中病毒蛋白与DNA之间的相互作用提供了一种简单方法。本章介绍了通过EMSA测量靶蛋白与DNA之间相互作用的详细信息和实验步骤。