|Entry||Database: EMDB / ID: 2132|
|Title||Cryo-electron tomography averaged map of microtubule doublet 2-8 in the distal/central region of Chlamydomonas flagella|
|Keywords||axoneme / dynein / Chlamydomonas / microtubule doublet / flagella|
|Sample||Outer doublets 2-8 of Chlamydomonas flagella in the distal/central region|
|Source||Chlamydomonas reinhardtii / plant / green algae|
|Map data||Reconstruction of the average of outer doublets 2-8 from Chlamydomonas flagella in the distal/central region.|
|Method||subtomogram averaging, at 36.8 Å resolution|
|Authors||Bui KH / Yagi T / Yamamoto R / Kamiya R / Ishikawa T|
|Citation||J. Cell Biol., 2012, 198, 913-925|
|Date||Deposition: Jun 11, 2012 / Header (metadata) release: Jun 13, 2012 / Map release: Sep 12, 2012 / Last update: Sep 26, 2012|
Downloads & links
|File||emd_2132.map.gz (map file in CCP4 format, 31251 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 7.25 Å|
CCP4 map header:
-Entire Outer doublets 2-8 of Chlamydomonas flagella in the distal/centra...
|Entire||Name: Outer doublets 2-8 of Chlamydomonas flagella in the distal/central region|
Details: Flagella were purified from Chlamydomonas / Number of components: 1
-Component #1: cellular-component, flagellum
|Cellular-component||Name: flagellum / a.k.a: axoneme, cilia / Recombinant expression: No|
|Source||Species: Chlamydomonas reinhardtii / plant / green algae / Strain: c137 (mt+)|
|Source (natural)||Organelle: flagella / Location in cell: distal part of the flagellum|
|Sample solution||Specimen conc.: 2 mg/ml|
Buffer solution: 30 mM Hepes, pH 7.4, 5 mM MgSO4, 1 mM DTT, 0.5 mM EDTA, 25 mM KCl and 0.5% (wt/vol) polyethylene glycol (MW 20,000)
|Support film||300 mesh Quantifoil Holey Carbon copper grid R2/1|
|Vitrification||Instrument: FEI VITROBOT MARK II / Cryogen name: ETHANE / Temperature: 110 K / Humidity: 90 % / Method: Offset -3, Blot 3s, Drain time 0s|
-Electron microscopy imaging
Model: Tecnai F20 / Image courtesy: FEI Company
|Imaging||Microscope: FEI TECNAI F20 / Date: Nov 26, 2010|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Electron dose: 60 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Magnification: 27500 X (nominal), 19303 X (calibrated) / Cs: 2 mm / Imaging mode: BRIGHT FIELD / Defocus: 4000 - 6000 nm / Energy filter: Gatan Tridiem / Energy window: 0-20 eV|
|Specimen Holder||Holder: Gatan 626 / Model: GATAN LIQUID NITROGEN / Tilt Angle: -60 - 60 deg. / Temperature: 98 K ( 93 - 118 K)|
|Camera||Detector: GATAN ULTRASCAN 1000 (2k x 2k)|
|Image acquisition||Number of digital images: 3770 / Bit depth: 16|
|Processing||Method: subtomogram averaging|
Details: R-weighted backprojection using IMOD with fiducial markers. Average number of tilts used in the 3D reconstructions: 61. Average tomographic tilt angle increment: 2.
|3D reconstruction||Algorithm: subtomogram averaging / Software: IMOD, BSOFT, SPIDER, TOM, package, Matlab / Resolution: 36.8 Å / Resolution method: FSC 0.5|
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