DOI: 10.22002/794j7-rcq33
title: JV3559_inPolara

description: Legionella thyA ΔdotG.This is a single mutant that lacks DotG.
description: <strong>Tilt Series Date:</strong> 2016-06-02</p>
<strong>Data Taken By:</strong> Debnath Ghosal</p>
<strong>Species / Specimen:</strong> Legionella pneumophila</p>
<strong>Strain:</strong> Philadelphia-1_Lp02</p>
<strong>Tilt Series Settings:</strong>  Single Axis, tilt range: (-60.0°, 60.0°), step: 1.5°, constant angular increment, dosage: 100.0 eV/Å²,  defocus: -6.0 μm,  magnification: 22500x. </p>
<strong>Microscope:</strong> Caltech Polara</p>
<strong>Acquisition Software:</strong> UCSFTomo</p>
<strong>Upload Method:</strong> pipeline</p>
<strong>Processing Software Used:</strong> Raptor</p>
<strong>Collaborators and Roles:</strong> Strain sent to us by Joseph P. Vogel, Washington University School of Medicine. Data collected and processed by Debnath Ghosal at F30, Caltech.</p>
<strong>Purification / Growth Conditions / Treatment:</strong> Cells were grown in AYE liquid culture with following supplements: ferric nitrate, cysteine hydrochloride and thymidine. Scrape cells off a plate, dissolve in the media, adjust the cell density to 0.6 and make a dilution series. Start O/N culture with all of them and freeze cells next day from the one in early stationary phase that is OD ~ 3.0. This time cell density was ~3.4</p>
<strong>Sample Preparation:</strong> 8 grids were frozen for the JV3559 strain at OD~ 3.4, 4 grids at density x and another 4 at density 2x. Vitrobot (MK iv) settings: 100% humidity, 4µl sample, Blot force 6, blot time 3 sec. Wait time 0 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200. Add gold to cells prior to freezing: 1ul of gold to 4 ul of cells, freeze.</p>
  dga2016-06-02-48_slicer16884.jpg: Beautiful double vesicle and seems like including ribosomes!!! dga2016-06-02-48_slicer16885.jpg: Beautiful double layered vesicle. I had another example where it included ribosomes. dga2016-06-02-48_slicer16886.jpg: Protein filament attaching it to the membrane?
description: Files available via S3 at https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-06-02-48/rawdata/</p>dg3559_020620160047.mrc,  Tilt Series (Pixel Size 4.8 nm), 2.3 GB  
        
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        dg3559_020620160047.rec,  Reconstruction (Pixel Size 19.2 nm), 890.9 MB  
        
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        3559.mod,  1.1 kB  
        
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        T4SS_2k.mod,  1.1 kB  
        
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        dg3559_020620160047.ali,  548.8 MB  
        
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        dg3559_020620160047.tlt,  616.0 B  
        
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        dg3559_020620160047_fid.tlt,  616.0 B  
        
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