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Team:UT Dallas/killswitch results
From 2011.igem.org
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<h2>PcstA+RBS+LuxI+double terminator</h2> | <h2>PcstA+RBS+LuxI+double terminator</h2> | ||
| - | This part produces LuxI,an enzyme for creating acyl-homoserine lactones from normal cell metabolites, in the absence of glucose. This part is repressed by glucose.<br> | + | This part produces LuxI,an enzyme for creating acyl-homoserine lactones from normal cell metabolites, in the absence of glucose. This part is repressed by glucose.<br><br> |
| - | <b>Experiment</b> | + | <b>Experiment</b><br> |
| - | We grew K569001 (Pcst-RBS-LuxI-terminator) and K131010 (AHL-inducible ColicinE2-GFP) transformed in BL21 E.coli cells in 3mL of LB broth with appropriate antibiotic overnight at 37C and 220rpm. Then we diluted the overnight samples 1:20 and allowed the cells to grow to a predetermined O.D. We combined the two cultures in various ratios and either added or did not add glucose. We allowed these samples to continue growing for 2 hours. After that, we took measurements using a fluorescent microscope. The images shown were the images used to get the data.<br> | + | We grew K569001 (Pcst-RBS-LuxI-terminator) and K131010 (AHL-inducible ColicinE2-GFP) transformed in BL21 E.coli cells in 3mL of LB broth with appropriate antibiotic overnight at 37C and 220rpm. Then we diluted the overnight samples 1:20 and allowed the cells to grow to a predetermined O.D. We combined the two cultures in various ratios and either added or did not add glucose. We allowed these samples to continue growing for 2 hours. After that, we took measurements using a fluorescent microscope. The images shown were the images used to get the data.<br><br> |
| - | All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.<br> | + | All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.<br><br> |
| - | These results show a correlation with glucose. The images without glucose show more fluorescence than the images with glucose<br> | + | These results show a correlation with glucose. The images without glucose show more fluorescence than the images with glucose<br><br> |
<img src="http://partsregistry.org/wiki/images/c/ca/No_glucose_pictures.png" width="720"><br> | <img src="http://partsregistry.org/wiki/images/c/ca/No_glucose_pictures.png" width="720"><br> | ||
Revision as of 22:29, 28 September 2011
PcstA+RBS+LuxI+double terminator
This part produces LuxI,an enzyme for creating acyl-homoserine lactones from normal cell metabolites, in the absence of glucose. This part is repressed by glucose.Experiment
We grew K569001 (Pcst-RBS-LuxI-terminator) and K131010 (AHL-inducible ColicinE2-GFP) transformed in BL21 E.coli cells in 3mL of LB broth with appropriate antibiotic overnight at 37C and 220rpm. Then we diluted the overnight samples 1:20 and allowed the cells to grow to a predetermined O.D. We combined the two cultures in various ratios and either added or did not add glucose. We allowed these samples to continue growing for 2 hours. After that, we took measurements using a fluorescent microscope. The images shown were the images used to get the data.
All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
These results show a correlation with glucose. The images without glucose show more fluorescence than the images with glucose
