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Yorodumi- EMDB-20438: Cryo-EM reconstruction of the mature chimeric BinJV/ZIKV-prME vir... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-20438 | |||||||||
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Title | Cryo-EM reconstruction of the mature chimeric BinJV/ZIKV-prME virion in complex with Fab C8 | |||||||||
Map data | Single particle reconstruction of the mature chimeric BinJV/ZIKV-prME virion complexed with Fab C8 | |||||||||
Sample |
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Biological species | Zika virus | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 12.2 Å | |||||||||
Authors | Watterson D | |||||||||
Funding support | Australia, 2 items
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Citation | Journal: Sci Transl Med / Year: 2019 Title: A recombinant platform for flavivirus vaccines and diagnostics using chimeras of a new insect-specific virus. Authors: Jody Hobson-Peters / Jessica J Harrison / Daniel Watterson / Jessamine E Hazlewood / Laura J Vet / Natalee D Newton / David Warrilow / Agathe M G Colmant / Carmel Taylor / Bixing Huang / ...Authors: Jody Hobson-Peters / Jessica J Harrison / Daniel Watterson / Jessamine E Hazlewood / Laura J Vet / Natalee D Newton / David Warrilow / Agathe M G Colmant / Carmel Taylor / Bixing Huang / Thisun B H Piyasena / Weng Kong Chow / Yin Xiang Setoh / Bing Tang / Eri Nakayama / Kexin Yan / Alberto A Amarilla / Sarah Wheatley / Peter R Moore / Mitchell Finger / Nina Kurucz / Naphak Modhiran / Paul R Young / Alexander A Khromykh / Helle Bielefeldt-Ohmann / Andreas Suhrbier / Roy A Hall / Abstract: Flaviviruses such as dengue, yellow fever, Zika, West Nile, and Japanese encephalitis virus present substantial global health burdens. New vaccines are being sought to address safety and ...Flaviviruses such as dengue, yellow fever, Zika, West Nile, and Japanese encephalitis virus present substantial global health burdens. New vaccines are being sought to address safety and manufacturing issues associated with current live attenuated vaccines. Here, we describe a new insect-specific flavivirus, Binjari virus, which was found to be remarkably tolerant for exchange of its structural protein genes (prME) with those of the aforementioned pathogenic vertebrate-infecting flaviviruses (VIFs). Chimeric BinJ/VIF-prME viruses remained replication defective in vertebrate cells but replicated with high efficiency in mosquito cells. Cryo-electron microscopy and monoclonal antibody binding studies illustrated that the chimeric BinJ/VIF-prME virus particles were structurally and immunologically similar to their parental VIFs. Pilot manufacturing in C6/36 cells suggests that high yields can be reached up to 10 cell culture infectious dose/ml or ≈7 mg/liter. BinJ/VIF-prME viruses showed utility in diagnostic (microsphere immunoassays and ELISAs using panels of human and equine sera) and vaccine applications (illustrating protection against Zika virus challenge in murine IFNAR mouse models). BinJ/VIF-prME viruses thus represent a versatile, noninfectious (for vertebrate cells), high-yield technology for generating chimeric flavivirus particles with low biocontainment requirements. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_20438.map.gz | 30.1 MB | EMDB map data format | |
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Header (meta data) | emd-20438-v30.xml emd-20438.xml | 11.3 KB 11.3 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_20438_fsc.xml | 13.7 KB | Display | FSC data file |
Images | emd_20438.png | 284.1 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-20438 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-20438 | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_20438.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Single particle reconstruction of the mature chimeric BinJV/ZIKV-prME virion complexed with Fab C8 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.027 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Zika virus
Entire | Name: Zika virus |
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Components |
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-Supramolecule #1: Zika virus
Supramolecule | Name: Zika virus / type: virus / ID: 1 / Parent: 0 / NCBI-ID: 64320 / Sci species name: Zika virus / Virus type: VIRION / Virus isolate: OTHER / Virus enveloped: Yes / Virus empty: No |
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Host system | Organism: Aedes albopictus (Asian tiger mosquito) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 2 mg/mL |
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Buffer | pH: 8 |
Grid | Support film - Material: CARBON / Support film - topology: LACEY / Details: unspecified |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK II |
Details | Gradient purified BinJV/ZIKV-prME |
-Electron microscopy
Microscope | FEI TECNAI F30 |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 26.7 e/Å2 |
Experimental equipment | Model: Tecnai F30 / Image courtesy: FEI Company |