Part:BBa_I13015

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:

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