Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria-2

<p><b>Copyright information:</b></p><p>Taken from "Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria"</p><p>http://www.biomedcentral.com/1471-2199/8/80</p><p>BMC Molecular Biology 2007;8():80-80.</p><p>Published online 19 Sep 2007</p><p>PMCID:PMC2039747.</p><p></p> ●; no sites) were grown to mid-log phase in Tris Minimal Succinate medium with 5 μg mlCm and 0.5% xylose. Bacteria were washed and resuspended in an equal volume of medium containing 5 μg mlCm and 1% glucose. Aliquots were placed in a 96 well microtitre plate and incubated at 37°C in a Tecan Genios Pro. Panel A: fluorescence (solid symbols, (Relative Fluorescence Units; RFU) and absorbance (open symbols) were measured at 10 min intervals. Plasmids used were pSB3004 (□, ■; 4 sites), pSB3002 (□, ▲; 2 sites) and pSB3000 (○, ●; no sites). Panel B: luminescence (solid symbols, Relative Light Units; RLU) and absorbance (open symbols) were measured at 10 minute intervals. Plasmids used were pSB3014 (□, ■; 4 sites), pSB3012 (□, ▲; 2 sites) and pSB3010 (○, ●; no sites). Data is presented as % maximal signal to allow direct comparison of repression kinetics despite the fact that light levels from each construct were different

Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria-0

<p><b>Copyright information:</b></p><p>Taken from "Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria"</p><p>http://www.biomedcentral.com/1471-2199/8/80</p><p>BMC Molecular Biology 2007;8():80-80.</p><p>Published online 19 Sep 2007</p><p>PMCID:PMC2039747.</p><p></p>; 4 sites) were grown in LB medium containing 0.5% (w/v) xylose at 37°C. Growth (OD 600 nm; open symbols) and luminescence (Relative Light Units; RLU closed symbols) were monitored over time. Reporter gene data are presented as RLU/ODto account for increasing cell number during the experiment

Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria-3

<p><b>Copyright information:</b></p><p>Taken from "Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria"</p><p>http://www.biomedcentral.com/1471-2199/8/80</p><p>BMC Molecular Biology 2007;8():80-80.</p><p>Published online 19 Sep 2007</p><p>PMCID:PMC2039747.</p><p></p>operon under the control of the Ppromoter (see Table 2). Cells were grown in MWB at 37°C and luminescence (closed symbols, Relative Light Units; RLU)) and growth measurements (open symbols) were taken at intervals. Reporter gene data are presented as RLU/ODto account for increasing cell number during the experiment

Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria-1

<p><b>Copyright information:</b></p><p>Taken from "Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria"</p><p>http://www.biomedcentral.com/1471-2199/8/80</p><p>BMC Molecular Biology 2007;8():80-80.</p><p>Published online 19 Sep 2007</p><p>PMCID:PMC2039747.</p><p></p> ●; no sites) were grown to mid-log phase in Tris Minimal Succinate medium with 5 μg mlCm and then diluted 1/20 into fresh medium supplemented with 0.5% xylose. Triplicate samples were placed in a 96 well microtitre plate and incubated at 37°C in a Tecan Genios Pro. Panel A: fluorescence (solid symbols, Relative Fluorescence Units; RFU) and absorbance (open symbols) were measured at 10 min intervals. Plasmids used were pSB3004 (□, ■; 4 sites), pSB3002 (□, ▲; 2 sites) and pSB3000 (○, ●; no sites). Panel B. luminescence (solid symbols, Relative Light Units; RLU)) and absorbance (open symbols) were measured at 10 min intervals. Plasmids used were pSB3014 (□, ■; 4 sites), pSB3012 (△, ▲; 2 sites) and pSB3010 (○, ●; no sites). Data is presented as % maximal signal to allow direct comparison of expression kinetics despite the fact that light levels from each construct were different