Difference between revisions of "Part:BBa J45995"
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===Usage and Biology===
 
===Usage and Biology===
 
See <partinfo>BBa_J45992</partinfo> for details on the ''osmY'' stationary phase promoter.
 
See <partinfo>BBa_J45992</partinfo> for details on the ''osmY'' stationary phase promoter.
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====William and Mary iGEM 2022====
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To test the effectiveness of our improved parts, our team grew the original MIT 2006 construct, our improved sfGFP construct, and our improved RFP construct in <i>E. coli</i> NEB5α in a plate reader. They were grown at 37°C using continuous shaking. For green fluorescence, we used an excitation value of 485 and an emission value of 528.  For red fluorescence, we used an excitation value of 584 and an emission value of 610. The values for green fluorescence are reported below. For information about red fluorescence, see parts page BBa_K4174001.
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https://static.igem.wiki/teams/4174/wiki/normalized-green-fluorescence.png
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As seen in the graph above, both the sfGFP and MIT GFP constructs enter stationary phase right before 14 hours, but our improved sfGFP circuit is much more fluorescent. The other constructs are our RFP construct and untransformed <i>E. coli</i> cells, both of which serve as negative controls for green fluorescence.
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https://static.igem.wiki/teams/4174/wiki/improveapart-smaller.png
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As seen in the image above, qualitative results reveal that our improved constructs are more fluorescent than the original construct. Here, sfGFP is on the far right, and is visibly more brightly green that the original GFP construct.
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Revision as of 18:48, 8 October 2022

Stationary phase dependent GFP generator

BBa_J45995 is a composite part consisting of an Escherichia coli osmY stationary phase promoter (BBa_J45992) and a GFP generator (BBa_E0840). Thus, BBa_J45995 produces fluorescence in stationary phase cultures.

Usage and Biology

See BBa_J45992 for details on the osmY stationary phase promoter.

William and Mary iGEM 2022

To test the effectiveness of our improved parts, our team grew the original MIT 2006 construct, our improved sfGFP construct, and our improved RFP construct in E. coli NEB5α in a plate reader. They were grown at 37°C using continuous shaking. For green fluorescence, we used an excitation value of 485 and an emission value of 528. For red fluorescence, we used an excitation value of 584 and an emission value of 610. The values for green fluorescence are reported below. For information about red fluorescence, see parts page BBa_K4174001.

normalized-green-fluorescence.png

As seen in the graph above, both the sfGFP and MIT GFP constructs enter stationary phase right before 14 hours, but our improved sfGFP circuit is much more fluorescent. The other constructs are our RFP construct and untransformed E. coli cells, both of which serve as negative controls for green fluorescence.


improveapart-smaller.png

As seen in the image above, qualitative results reveal that our improved constructs are more fluorescent than the original construct. Here, sfGFP is on the far right, and is visibly more brightly green that the original GFP construct.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 872