Difference between revisions of "Part:BBa K4591002"
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<p><i>XylS</i> is an archetype transcriptional activator of AraC/XylS family, mined from the TOL plasmid pWW0 of the bacterium Pseudomonas putida. It is composed of a C-terminal domain (CTD) involved in DNA binding, and an N-terminal domain required for effector binding an`d protein dimerization. [1]</p> | <p><i>XylS</i> is an archetype transcriptional activator of AraC/XylS family, mined from the TOL plasmid pWW0 of the bacterium Pseudomonas putida. It is composed of a C-terminal domain (CTD) involved in DNA binding, and an N-terminal domain required for effector binding an`d protein dimerization. [1]</p> | ||
<p>XylS can bind benzoic acid and various derivatives, | <p>XylS can bind benzoic acid and various derivatives, | ||
| − | but it cannot recognize PA and TPA.[2-5] So the Jiawei Li and Mario Roque Huanca Nina successfully make the directed evolution of XylS to generate new TFs that are capable of binding and responding to TPA and PA . Such XylS mutants could be used to construct whole-cell biosensors for fluorimetric detection of PA and TPA. </p> | + | but it cannot recognize PA and TPA.[2-5] So the Jiawei Li and Mario Roque Huanca Nina successfully make the directed evolution of XylS to generate new TFs that are capable of binding and responding to TPA and PA. Such XylS mutants could be used to construct whole-cell biosensors for fluorimetric detection of PA and TPA. </p> |
| − | + | <p>When the TPA was detected by the Xylsmut, the P<sub>m</sub> promotor would be actived and transcript the downstream fragments.[2-5,6]</p> | |
| + | <p>Depends on this, previews researchers have construct a simple XylSmut-based fluorometric biosensors which can detect the TPA.It contains a Pm promotor, a constitutive promotor, a fluorometric protein such as the GFP, and a terminator. [7]</p> | ||
<div style="display: flex;justify-content:center;align-items:center;flex-direction: column;"> | <div style="display: flex;justify-content:center;align-items:center;flex-direction: column;"> | ||
<div> | <div> | ||
<img src="https://static.igem.wiki/teams/4591/wiki/parts/tpa.svg" width="900" height="auto"> | <img src="https://static.igem.wiki/teams/4591/wiki/parts/tpa.svg" width="900" height="auto"> | ||
</div> | </div> | ||
| − | <p style="text-align: center;">Fig 1. | + | <p style="text-align: center;">Fig 1. A kind of simple XylSmut-based fluorometric biosensors which can detect the TPA</p> |
</div> | </div> | ||
| − | + | ||
<p>In this year, we have designed a system of PET detection, attachment and degradation.In order to detect the TPA, the degradation products of the PET. The Xylsmut was chosen as the "sensor" of the TPA concentration, and the deGFP was also constructed in our circuit as the indicator of TPA.</p> | <p>In this year, we have designed a system of PET detection, attachment and degradation.In order to detect the TPA, the degradation products of the PET. The Xylsmut was chosen as the "sensor" of the TPA concentration, and the deGFP was also constructed in our circuit as the indicator of TPA.</p> | ||
</html> | </html> | ||
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https://static.igem.wiki/teams/4591/wiki/parts/2023xylsmut-pig2.jpg | https://static.igem.wiki/teams/4591/wiki/parts/2023xylsmut-pig2.jpg | ||
</div> | </div> | ||
| − | <p style="text-align: center;"> | + | <p style="text-align: center;"><sup>[6]</sup></p> |
</div> | </div> | ||
Revision as of 09:21, 8 October 2023
XylSmut
iGEM23_ZJUT-China's Contribution
Improvement
The xylsmut is upgraded form an existing part Xyls(BBa_K108029)from the Team iGEM_Tsinghua.It was
Usage and Biology
XylS is an archetype transcriptional activator of AraC/XylS family, mined from the TOL plasmid pWW0 of the bacterium Pseudomonas putida. It is composed of a C-terminal domain (CTD) involved in DNA binding, and an N-terminal domain required for effector binding an`d protein dimerization. [1]
XylS can bind benzoic acid and various derivatives, but it cannot recognize PA and TPA.[2-5] So the Jiawei Li and Mario Roque Huanca Nina successfully make the directed evolution of XylS to generate new TFs that are capable of binding and responding to TPA and PA. Such XylS mutants could be used to construct whole-cell biosensors for fluorimetric detection of PA and TPA.
When the TPA was detected by the Xylsmut, the Pm promotor would be actived and transcript the downstream fragments.[2-5,6]
Depends on this, previews researchers have construct a simple XylSmut-based fluorometric biosensors which can detect the TPA.It contains a Pm promotor, a constitutive promotor, a fluorometric protein such as the GFP, and a terminator. [7]
Fig 1. A kind of simple XylSmut-based fluorometric biosensors which can detect the TPA
In this year, we have designed a system of PET detection, attachment and degradation.In order to detect the TPA, the degradation products of the PET. The Xylsmut was chosen as the "sensor" of the TPA concentration, and the deGFP was also constructed in our circuit as the indicator of TPA.
Characterization
(The following experimental data are from the literature and modeling, not the actual data made by the experiment)
Fig 2. The time-course fluorescence changes of pUC57-XylS-sfGFP in response to different inducers.
a) 2-hydroxybenzoic acid, b) 3-chlorobenzoic acid, c) PA, and d) TPA.[6]
[6]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]