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- W1966738863 abstract "Cryo-electron tomography (cryoET) has become a powerful tool for direct visualization of 3D structures of native biological specimens at molecular resolution, but its application is limited to thin specimens (<300 nm). Recently, vitreous sectioning and cryoFIB milling technologies were developed to physically reduce the specimen thickness; however, cryoET analysis of membrane protein complexes within native cell membranes remains a great challenge. Here, we use phage ΦX174 lysis gene E to rapidly produce native, intact, bacterial cell membranes for high resolution cryoET. We characterized E gene-induced cell lysis using FIB/SEM and cryoEM and showed that the bacteria cytoplasm was largely depleted through spot lesion, producing ghosts with the cell membranes intact. We further demonstrated the utility of E-gene-induced lysis for cryoET using the bacterial chemotaxis receptor signaling complex array. The described method should have a broad application for structural and functional studies of native, intact cell membranes and membrane protein complexes." @default.
- W1966738863 created "2016-06-24" @default.
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- W1966738863 date "2014-12-01" @default.
- W1966738863 modified "2023-10-18" @default.
- W1966738863 title "Controlled Bacterial Lysis for Electron Tomography of Native Cell Membranes" @default.
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- W1966738863 doi "https://doi.org/10.1016/j.str.2014.09.017" @default.
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