Difference between revisions of "Part:BBa K3225014"
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<partinfo>BBa_K3225014 short</partinfo> | <partinfo>BBa_K3225014 short</partinfo> | ||
| − | This device is designed to detect N-(3-oxohexanoyl)-L-homoserine lactone (OHHL | + | This device is designed to detect N-(3-oxohexanoyl)-L-homoserine lactone (OHHL https://2019.igem.org/wiki/images/0/0c/T--iBowu-China--DesignOHHL.png , |
quorum sensing signal molecular of E.carotovora). It consists of LuxR(quorum sensing transcriptional factor, BBa_C0062) drove by a constitutive promoter J23101 and GFP reporter drove by the binding promoter of LuxR(pLux, BBa_R0062). | quorum sensing signal molecular of E.carotovora). It consists of LuxR(quorum sensing transcriptional factor, BBa_C0062) drove by a constitutive promoter J23101 and GFP reporter drove by the binding promoter of LuxR(pLux, BBa_R0062). | ||
Revision as of 17:02, 21 October 2019
J23101-LuxR-pLux-GFP
This device 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 GFP reporter drove by the binding promoter of LuxR(pLux, BBa_R0062).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1037
Results
1.Results in vivo
From figure1, we found that the GFP intensity increased after OHHL incubation, even 1nM OHHL, which means LuxR could bind to OHHL efficiently. It showed that LuxR - pLux could be a highly sensitive detector of OHHL.
Figure 1.The fluorescence intensity over the time with different concentration of OHHL treatment. The inducer OHHL was added to the culture after 60 min incubation.
2.Results in vitro(cell-free system).
After the cellular sensing characterization, we tested the binding efficiency of LuxR to OHHL in cell-free systems(Fig. 3). After the overnight cell-free incubation, it showed that the circuit of LuxR-GFP was responsible to AHL with the detection limit of 1 nM.
Figure 2. The cell-free fluorescence output induced by a series of AHL concentrations. FLU was measured after overnight incubation