DOI: 10.22002/vqs0e-68691
title: E.coli co-overexpression

description: E.coli co overexpression UU1607/pJC3/pPM25
description: <strong>Tilt Series Date:</strong> 2013-07-17</p>
<strong>Data Taken By:</strong> Ariane Briegel</p>
<strong>Species / Specimen:</strong> Escherichia coli</p>
<strong>Strain:</strong> RP 437</p>
<strong>Tilt Series Settings:</strong>  Single Axis, tilt range: (-66.0°, 66.0°), step: 2°, constant angular increment, dosage: 160.0 eV/Å²,  defocus: -3.0 μm,  magnification: 41000x. </p>
<strong>Microscope:</strong> Caltech Polara</p>
<strong>Acquisition Software:</strong> UCSFTomo</p>
<strong>Upload Method:</strong> reupload_downloaded</p>
<strong>Processing Software Used:</strong> SIRT</p>
<strong>Collaborators and Roles:</strong> strain sent from Heather Hodges (Kiessling lab), parent strain and plasmids published from the Parkinson lab</p>
<strong>Purification / Growth Conditions / Treatment:</strong> Grew UU1607/pJC3/pPM25 overnight in 5mL TB + 3.8µl Cm + 5µl CarbDiluted back 1:50 into 200mL TB + 152µl Cm + 200µl Carb + 50µl 1M IPTG + 200µl Na-salicylateGrew ~2 hours to mid-log phaseSpun down 10' at 5,000 x gResuspended in 300µl lysis buffer + 0.1µl PMSF + 0.75 µl 1,10-phenanthrolineAdded 33.3µl (2mg/ml) fresh lysozyme solution (20mg/ml in 10mM Tris-HCl, pH 8.0)Incubated 45' at 37°CAdded 1.67µl 1M MgCl2, 0.2µl 1M CaCl2, 0.33µl DNase IIncubated 30' at 37°CSpun down 4000 g 5 min Resuspended in activity buffer (cold room) without glycerol</p>
description: Files available via S3 at https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/aba2013-07-17-34/rawdata/</p>cooverexpressionatlas4micronvarioussangles0004.mrc,  Tilt Series (Pixel Size 0.28 nm), 1.9 GB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/aba2013-07-17-34/rawdata/cooverexpressionatlas4micronvarioussangles0004.mrc"
        > <i class="download icon"></i> Download </a></p>

        cooverexpressionatlas4micronvarioussangles0004SIRTrot.rec,  Reconstruction (Pixel Size 0.0 nm), 11.4 GB  
        
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        > <i class="download icon"></i> Download </a></p>

        cooverexpressionatlas4micronvarioussanglesminusten0004.rec,  Reconstruction (Pixel Size 0.0 nm), 3.6 GB  
        
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        > <i class="download icon"></i> Download </a></p>

        cooverexpressionatlas4micronvarioussanglesminusten0004SIRTrot.rec,  Reconstruction (Pixel Size 0.0 nm), 14.2 GB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/aba2013-07-17-34/3dimage_46582/cooverexpressionatlas4micronvarioussanglesminusten0004SIRTrot.rec"
        > <i class="download icon"></i> Download </a></p>

        cooverexpressionatlas4micronvarioussanglesminusten0004SIRTrotbin.rec,  Reconstruction (Pixel Size 0.0 nm), 1.8 GB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/aba2013-07-17-34/3dimage_46584/cooverexpressionatlas4micronvarioussanglesminusten0004SIRTrotbin.rec"
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        SIRTrot.rec,  Reconstruction (Pixel Size 0.0 nm), 14.2 GB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/aba2013-07-17-34/3dimage_46585/SIRTrot.rec"
        > <i class="download icon"></i> Download </a></p>

        SIRTrotbinmedian.rec,  Reconstruction (Pixel Size 0.0 nm), 1.8 GB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/aba2013-07-17-34/3dimage_46588/SIRTrotbinmedian.rec"
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