Part:BBa_K3225015

J23101-LuxR-pLux-LacZ

This gene circuit is designed to detect N-(3-oxohexanoyl)-L-homoserine lactone (OHHL ，quorum sensing signal molecular of E.carotovora). It consists of LuxR(quorum sensing transcriptional factor, BBa_C0062) drove by a constitutive promoter J23101 and LacZ(BBa_I732017) as a repoter drove by the binding promoter of LuxR(pLux, BBa_R0062).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Results

High-sensitivity OHHL test paper

The gene circuits coupling with cell-free expression system are lyophilized on paper by freeze-drying technology. The reporter is LacZ which is β-galactosidase which can cleave the yellow substrate to purple. OHHL can be detected after soaking the test paper with water.

Figure 1. Real shot of OHHL test strip

Among the series of AHL concentrations induction, it showed that the detection limit was lower than 1 nM. Moreover, there is a significant visible output with 10 nm AHL treatment (Fig. 2).

Figure 2. The detection limit and detection time of quorum sensing based testing strips.