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呼吸道合胞病毒蛋白复合物的单颗粒电子显微镜分析

Single-Particle Electron Microscopy Analysis of Respiratory Syncytial Virus Protein Complexes.

作者信息

Cao Dongdong, Gao Yunrong, Chen Zhenhang, Horgan Natalie, Liang Bo

机构信息

Department of Biochemistry, Emory University School of Medicine, Atlanta, GA, USA.

出版信息

Methods Mol Biol. 2025;2948:159-180. doi: 10.1007/978-1-0716-4666-3_11.

Abstract

Cryogenic electron microscopy (cryo-EM) has been an important technique for determining the structures of macromolecules at near-atomic resolution in their native states. In recent years, the structures of some respiratory syncytial virus (RSV) protein complexes, such as the RSV polymerase (L:P complex) and nucleoprotein-RNA complex, have been determined by cryo-EM, providing insights into the mechanisms of viral RNA synthesis and facilitating antiviral development. Here, we describe the step-by-step protocols for the negative-stain and cryo-EM analysis of the RSV protein complexes.

摘要

低温电子显微镜(cryo-EM)一直是在天然状态下以近原子分辨率测定大分子结构的重要技术。近年来,一些呼吸道合胞病毒(RSV)蛋白复合物的结构,如RSV聚合酶(L:P复合物)和核蛋白-RNA复合物,已通过低温电子显微镜确定,这为深入了解病毒RNA合成机制和促进抗病毒药物研发提供了线索。在此,我们描述了对RSV蛋白复合物进行负染和低温电子显微镜分析的详细步骤方案。

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