Team:UT Dallas/killswitch results

From 2011.igem.org

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<h2>Results</h2>
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<partinfo>BBa_K569001 short</partinfo>
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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>
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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.  
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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>
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The results show that, as planned, the removal of glucose from the system results in widespread death of both the strains.<br>
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<img src="http://partsregistry.org/wiki/images/c/ca/No_glucose_pictures.png" width="720"><br><br>
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<img src="http://partsregistry.org/wiki/images/9/90/Without_glucose.png" width="720"><br>
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===Usage and Biology===
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<img src="http://partsregistry.org/wiki/images/6/6c/Glucose_pictures_2.png" width="720"><br><br>
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<img src="http://partsregistry.org/wiki/images/d/d3/With_glucose2.png" width="720"><br>
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K569001 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K569001 parameters</partinfo>
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'''Experiment'''
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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.
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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.
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These results show a correlation with glucose. The images without glucose show more fluorescence than the images with glucose.
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<img src="http://partsregistry.org/wiki/images/c/ca/No_glucose_pictures.png" width="900">
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<img src="http://partsregistry.org/wiki/images/9/90/Without_glucose.png" width="750">
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<img src="http://partsregistry.org/wiki/images/6/6c/Glucose_pictures_2.png" width="900">
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<img src="http://partsregistry.org/wiki/images/d/d3/With_glucose2.png" width="750">
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Latest revision as of 01:06, 29 September 2011

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