Difference between revisions of "Part:BBa K131010:Experience"

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===Applications of BBa_K131010===
 
===Applications of BBa_K131010===
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'''Team UT_Dallas 2011'''
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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. They also show that the colicin E2 is causing apoptosis by the decrease in fluorescence when there is more Pcst-RBS-LuxI-terminator, which means there is more AHL to activate the AHL-inducible ColicinE2-GPF.
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<img src="https://static.igem.org/mediawiki/parts/c/ca/No_glucose_pictures.png" width="900">
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<img src="https://static.igem.org/mediawiki/parts/9/90/Without_glucose.png" width="900">
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<img src="https://static.igem.org/mediawiki/parts/6/6c/Glucose_pictures_2.png" width="900">
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<img src="https://static.igem.org/mediawiki/parts/d/d3/With_glucose2.png" width="900">
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===User Reviews===
 
===User Reviews===

Latest revision as of 22:16, 28 September 2011

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Applications of BBa_K131010

Team UT_Dallas 2011

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. They also show that the colicin E2 is causing apoptosis by the decrease in fluorescence when there is more Pcst-RBS-LuxI-terminator, which means there is more AHL to activate the AHL-inducible ColicinE2-GPF.

===User Reviews=== BBa_K131010 StartReviews BBa_K131010 EndReviews '''Sequence analysis of 2009 distributed BioBrick''' This part has been classified as "LONG PART" by iGEM QC, but we noticed that the 3' end of this part is not correct: it does not contain BBa_K131009.