Difference between revisions of "Part:BBa K569001"
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Experiment
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'''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.  
 
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.  

Revision as of 22:17, 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.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 794
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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.