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We cultivate the recevier E.coli until the OD up to 0.4 and add salicylateinto 16 EP tube containing aforementioned E.coli respectively. Different concentration of salicylate solution will be added, that is 10-7 M, 10-6 M, 5×10-6 M, 10-5 M, 2.5×10-5 M, 5×10-5 M, 7.5×10-5 M, 10-4 M, 2.5×10-4 M, 5×10-4 M, 7.5×10-4 M, 10-3 M. Cultivate the cell for 3 hours at 37℃. Then centrifuged cells and resuspended in phosphate buffer solution (PBS). The GFP fluorescence of each culture was obtained using FCM(flow cytometry). The result is shown in figure 1.
 
We cultivate the recevier E.coli until the OD up to 0.4 and add salicylateinto 16 EP tube containing aforementioned E.coli respectively. Different concentration of salicylate solution will be added, that is 10-7 M, 10-6 M, 5×10-6 M, 10-5 M, 2.5×10-5 M, 5×10-5 M, 7.5×10-5 M, 10-4 M, 2.5×10-4 M, 5×10-4 M, 7.5×10-4 M, 10-3 M. Cultivate the cell for 3 hours at 37℃. Then centrifuged cells and resuspended in phosphate buffer solution (PBS). The GFP fluorescence of each culture was obtained using FCM(flow cytometry). The result is shown in figure 1.
  
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''Figure 3. Salicylate receiver system cell distribution of GFP fluorescence intensity.''
 
''Figure 3. Salicylate receiver system cell distribution of GFP fluorescence intensity.''
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Latest revision as of 20:03, 18 October 2011

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User Reviews

UNIQ95a6197b36896d7c-partinfo-00000000-QINU UNIQ95a6197b36896d7c-partinfo-00000001-QINU

No review score entered. Peking_S iGEM 2011

We cultivate the recevier E.coli until the OD up to 0.4 and add salicylateinto 16 EP tube containing aforementioned E.coli respectively. Different concentration of salicylate solution will be added, that is 10-7 M, 10-6 M, 5×10-6 M, 10-5 M, 2.5×10-5 M, 5×10-5 M, 7.5×10-5 M, 10-4 M, 2.5×10-4 M, 5×10-4 M, 7.5×10-4 M, 10-3 M. Cultivate the cell for 3 hours at 37℃. Then centrifuged cells and resuspended in phosphate buffer solution (PBS). The GFP fluorescence of each culture was obtained using FCM(flow cytometry). The result is shown in figure 1.

LM7.png

Figure 1. Dose response curve. Horizontal axis represents GFP expression measured by flow cytometry and the vertical axis represen different salicylate concentration. Data was fitted with Hill function.

salicylate system

We cultivate the recevier E.coli until its OD up to 0.4. Afterward, add salicylate into the liquid medium and salicylate ultimate concentration is 10-4 M, at last the gfp expression level of different length will be measured by FCM(flow cytometry), delta time is 15 min or 30 min between each sample.

LM10.png

Figure 2. Time response curve of salicylate recevier. Horizontal axis represent time while the vertical axis represent average fluorescence intensity measured by flow cytometry.

Coordinatiblity

LM12b.png

Figure 3. Salicylate receiver system cell distribution of GFP fluorescence intensity.

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