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Title:Cryo-electron tomography of in situ flagellar motor of Borrelia burgdorferi FlgE mutant
Authors:Zhao X, Zhang K, Boquoi T, Hu B, Motaleb MA, Miller KA, James ME, Charon NW, Manson MD, Norris SJ, Li C, Liu J
Sample:In situ flagellar motor from B. burgdorferi FlgE mutant cells
Method:Subtomogram averaging (38 angstroms resolution)
Related entries in frame:EMD-5627, EMD-5628, EMD-5629, EMD-5630, EMD-5631, EMD-5632
Red flagLatest update:2014-03-26
Sample
Sample name: In situ flagellar motor from B. burgdorferi FlgE mutant cells
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1cellular-componentflagellar motorfalseBorrelia burgdorferi
Experiment
Specimen state: Cell
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
mg/mLBSK-II liquid medium supplemented with 6% rabbit serumfreshly glow-discharged holey carbon grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGERBacterial culture was deposited onto freshly glow-discharged, holey carbon grids for 1 min. Grids were blotted with filter paper and then rapidly frozen in liquid ethane, using a homemade gravity-driven plunger apparatus. ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVFLOOD BEAMBRIGHT FIELD2 mm6000 nm8000 nm31000FIELD EMISSION GUN mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
OTHER-65°65° eV K170 K K mrad100 e/Å201-NOV-2012
Processing
Protocol:subvolume averaging
Software:IMOD, Raptor, Protomo
Number of subtomogram averaged:999
Number of class averages:4
Imposed symmetry:C1
Resolution by author:38 Å
Resolution method:FSC 0.5
Processing details:The subvolumes of the flagellar motors were extracted computationally from the tomograms and were further aligned in 3D.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
9831 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails