Using cryoEM and cryoET to visualize membrane penetration of a non-enveloped virus

利用冷冻电镜和冷冻电镜断层扫描技术观察无包膜病毒的膜穿透过程

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作者:Xian Xia ,Z Hong Zhou

Abstract

Key to cell entry by non-enveloped viruses is virus-cell interactions at the cell or endosomal membrane. Here, we detail our protocols to capture such interactions between non-enveloped virus bluetongue virus (BTV) and vesicular membrane by cryogenic electron microscopy (cryoEM) and tomography (cryoET). Key steps include virus isolation, liposome preparation, virus-liposome incubation and vitrification, cryoEM and cryoET imaging, data processing for 3D reconstruction, and subtomogram averaging. The protocols can be generally applicable to studies of cell entry by other non-enveloped viruses. For complete details on the use and execution of this protocol, please refer to Xia et al. (2021). Keywords: Cryo-EM; Microbiology; Microscopy; Structural biology.

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