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Title:Cryo-EM structure of HBV T=4 pgRNA-filled Cp183 capsid
Authors:Wang JC-Y, Dhasan RS, Zlotnick A
Sample:HBV T=4 pgRNA-filled Cp183 capsid
Method:Single particle reconstruction (8.0 angstroms resolution)
Red flagLatest update:2013-07-24
Sample
Sample name: HBV T=4 pgRNA-filled Cp183 capsid
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1virusHepatitis B virusHepatitis B virusReassembled the purified Cp183 dimers with in vitro transcribed HBV pgRNA at a molar ratio of protein dimer to RNA polymer = 120:1
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.5 mg/mL150 mM NaCl, 50 mM HEPES, 2 mM DTTQuantifoil R 2/2 holey carbon 200 mesh copper grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100%93 KFEI VITROBOT MARK IIIBlot for 4 seconds before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 3200FS300 kVFLOOD BEAMBRIGHT FIELD1.1 mm270 nm3800 nm40000FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k) mmobjective lens astigmatism was corrected at 80,000 times magnification
JEOL 3200FS300 kVFLOOD BEAMBRIGHT FIELD1.1 mm270 nm3800 nm40000FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k) mmobjective lens astigmatism was corrected at 80,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side entry liquid nitrogen-cooled cryo specimen holderGATAN LIQUID NITROGEN°°Omega filter eV97 K K K mrad14 e/Å227-APR-2010
Side entry liquid nitrogen-cooled cryo specimen holderGATAN LIQUID NITROGEN°°Omega filter eV97 K K K mrad14 e/Å204-MAY-2010
Processing
Software:Auto3dem
CTF correction:Each particle phase-flipping
Number of particles:7201
Imposed symmetry:I
Resolution by author:8.0 Å
Resolution method:FSC 0.5
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
193 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails