Cao Dongdong, Gao Yunrong, Chen Zhenhang, Horgan Natalie, Liang Bo
Department of Biochemistry, Emory University School of Medicine, Atlanta, GA, USA.
Methods Mol Biol. 2025;2948:159-180. doi: 10.1007/978-1-0716-4666-3_11.
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.
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。