Difference between revisions of "Part:BBa K4591002"
| Line 41: | Line 41: | ||
XylS mutants in response to PA and TPA. (a) XylS-K38R-L224Q, PA; (b) XylS-K38R-L224Q, TPA; (c) XylS-W88C-L224Q, PA; (d) W88C-L224Q, TPA. <sup>[6]</sup></p> | XylS mutants in response to PA and TPA. (a) XylS-K38R-L224Q, PA; (b) XylS-K38R-L224Q, TPA; (c) XylS-W88C-L224Q, PA; (d) W88C-L224Q, TPA. <sup>[6]</sup></p> | ||
</div> | </div> | ||
| − | + | <p>The data shows that the TF XylS<sub>K38R-L224Q</sub> have higher sensitivity of the TPA we choose the XylS<sub>K38R-L224Q</sub> as our new part, Xylsmut.</p> | |
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K4591002 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4591002 SequenceAndFeatures</partinfo> | ||
Revision as of 17:32, 8 October 2023
XylSmut
iGEM23_ZJUT-China's Contribution
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 and 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. Among these new TFs, XylsmutK32R-L224Q and XylsmutW88C-L224Q have showed the capacity of binding and responding to TPA and PA. Such XylS mutants could be used to construct whole-cell biosensors for fluorometric 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)
Limited by the Labotory equipment, some of the data was collected from the literature.
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]
The time-course fluorescence of whole-cell biosensors containing different XylS mutants in response to PA and TPA. (a) XylS-K38R-L224Q, PA; (b) XylS-K38R-L224Q, TPA; (c) XylS-W88C-L224Q, PA; (d) W88C-L224Q, TPA. [6]
The data shows that the TF XylSK38R-L224Q have higher sensitivity of the TPA we choose the XylSK38R-L224Q as our new part, Xylsmut.
Sequence and Features Status: 500 Content-type: text/html
Software error:
Can't call method "errstr" on an undefined value at /websites/parts.igem.org/cgi/lib/BBDB.pm line 35. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Plate.pm line 13. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Plate.pm line 13. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Well.pm line 11. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Well.pm line 11. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Blast.pm line 15. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Blast.pm line 15. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Sample.pm line 24. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Sample.pm line 24. Compilation failed in require at /websites/parts.igem.org/cgi/lib/WrapPart.pm line 10. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/WrapPart.pm line 10. Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 8. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.
For help, please send mail to this site's webmaster, giving this error message and the time and date of the error.