Difference between revisions of "Part:BBa I13015"
 
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<h4>Activity compared to synthetic 3OC6 HSL</h4>
 
<h4>Activity compared to synthetic 3OC6 HSL</h4>
  
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Team USP-Brazil (2018) characterized this part by using it in a quorum sensing system along with LuxR and promoter pLux (see [https://parts.igem.org/Part:BBa_K2771030 BBa_K2771030]), in an assay comparing pBad at full activity (2.5mM arabinose) and a quantity known to activate well pLux (10<sup>-4</sup> mM HSL):
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Team USP-Brazil (2018) characterized this part by using it in a quorum sensing system along with LuxR and promoter pLux (see [https://parts.igem.org/Part:BBa_K2771030 BBa_K2771030]), in an assay comparing pBad at full activity (2.5mM arabinose) and a quantity known to activate well pLux (10<sup>-4</sup> mM HSL). Arabinose induction showed an equilibrium about 7x higher than with just synthetic HSL, and a bigger EC50, due to the necessity of transcription of two genes and signal production by LuxI for the reporter to be seeable:
  
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[[File:T--USP-Brazil--Lux_Las_assay.png|500px|thumb|none|alt=experiment 1.|Figure 2: Liquid culture assay with 2.5 mM arabinose or 10<sup>-4</sup> mM 3OC6 HSL induction at 0h. Arabinose induction shows clearly higher concentration of HSL than in the synthetic HSL curve, showing higher curve inclination and higher equilibrium point]]
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[[File:T--USP-Brazil--ara HSL assay.png|500px|thumb|none|alt=experiment 1.|Figure 2: Liquid culture assay with 2.5 mM arabinose or 10<sup>-4</sup> mM 3OC6 HSL induction at 0h. Arabinose induction shows clearly higher concentration of HSL than in the synthetic HSL curve, showing higher curve inclination and higher equilibrium point. Fluorescence measured as a ratio of a YFP reporter with the quorum sensing promoter, normalized by the value of a constitutive CFP reporter, proxy for cell conditions and environmental influence]]

Latest revision as of 17:43, 17 October 2018


3OC6HSL Sender controlled by pBad

The pBad promoter controlling a LuxI sender device. The protein is an untagged LuxI enzyme which produces an AHL (3OC6HSL).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1873
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Activity compared to synthetic 3OC6 HSL

Team USP-Brazil (2018) characterized this part by using it in a quorum sensing system along with LuxR and promoter pLux (see BBa_K2771030), in an assay comparing pBad at full activity (2.5mM arabinose) and a quantity known to activate well pLux (10-4 mM HSL). Arabinose induction showed an equilibrium about 7x higher than with just synthetic HSL, and a bigger EC50, due to the necessity of transcription of two genes and signal production by LuxI for the reporter to be seeable:

experiment 1.
Figure 2: Liquid culture assay with 2.5 mM arabinose or 10-4 mM 3OC6 HSL induction at 0h. Arabinose induction shows clearly higher concentration of HSL than in the synthetic HSL curve, showing higher curve inclination and higher equilibrium point. Fluorescence measured as a ratio of a YFP reporter with the quorum sensing promoter, normalized by the value of a constitutive CFP reporter, proxy for cell conditions and environmental influence