Difference between revisions of "Part:BBa K3225015"
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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. | 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. | ||
<img style="width:800px;"src="https://static.igem.org/mediawiki/parts/6/64/T--iBowu-China--Character-5015-product-v2.png"> | <img style="width:800px;"src="https://static.igem.org/mediawiki/parts/6/64/T--iBowu-China--Character-5015-product-v2.png"> | ||
| − | <p>Figure 1. Real shot of OHHL test | + | <p>Figure 1. Real shot of OHHL test paper.</p> |
<p>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).</p> | <p>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).</p> | ||
<img style="width:800px;"src="https://static.igem.org/mediawiki/parts/8/83/T--iBowu-China--Character-5015-fig1.jpeg"> | <img style="width:800px;"src="https://static.igem.org/mediawiki/parts/8/83/T--iBowu-China--Character-5015-fig1.jpeg"> | ||
Latest revision as of 21:03, 21 October 2019
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:
- 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]
- 1000COMPATIBLE 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 paper.
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.