Difference between revisions of "Part:BBa K2507015"
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==Characterization== | ==Characterization== | ||
We first validated that this system can function as a tetrathionate sensor and reporter in the laboratory strains <i>Escherichia coli</i> Top10 and <i>E. coli<.i> Nissle 1917. | We first validated that this system can function as a tetrathionate sensor and reporter in the laboratory strains <i>Escherichia coli</i> Top10 and <i>E. coli<.i> Nissle 1917. | ||
| − | [[File: SHSBNU 17 | + | [[File: SHSBNU 17 40a12.jpg|600px|center|Figure1]] |
Figure 1. Schematic diagram of ligand-induced signaling through TtrS/R and plasmid-borne implementation of the sensor components. TtrS/R was tested with BBa_K2507006 integrated into the pSB4K5 backbone and BBa_K2507015 into the pSB1C3 backbone. We submitted all parts to the iGEM registry in pSB1C3. | Figure 1. Schematic diagram of ligand-induced signaling through TtrS/R and plasmid-borne implementation of the sensor components. TtrS/R was tested with BBa_K2507006 integrated into the pSB4K5 backbone and BBa_K2507015 into the pSB1C3 backbone. We submitted all parts to the iGEM registry in pSB1C3. | ||
Revision as of 13:27, 1 November 2017
J23109-ttrR-PttrB185-BBa_K1033932
Usage and Biology
E. coli-codon-optimized TtrS(BBa_K2507002) and TtrR (BBa_K2507003) are two basic parts which are derived from the two-component system of the marine bacterium Shewanella baltica. TtrS is the membrane-bound sensor kinase (SK) which can sense tetrathionate outside the cell, and TtrR is the DNA-binding response regulator (RR). PttrB185-269 (BBa_K2507019) is a minimal TtrR-activated promoter which is activated when TtrR is phosphorylated by TtrS after TtrS senses tetrathionate.
Winter et al. have shown that reactive oxygen species (ROS) produced by the host during inflammation convert thiosulfate into tetrathionate, which this pathogen consumes to establish a beachhead for infection (Winter et al, 2010). Thus, tetrathionate may correlate with pro-inflammatory conditions and can therefore be used as a sensor for intestinal inflammation.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 11
Illegal NheI site found at 34 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 318
- 1000COMPATIBLE WITH RFC[1000]
Characterization
We first validated that this system can function as a tetrathionate sensor and reporter in the laboratory strains Escherichia coli Top10 and E. coli<.i> Nissle 1917.
Figure 1. Schematic diagram of ligand-induced signaling through TtrS/R and plasmid-borne implementation of the sensor components. TtrS/R was tested with BBa_K2507006 integrated into the pSB4K5 backbone and BBa_K2507015 into the pSB1C3 backbone. We submitted all parts to the iGEM registry in pSB1C3.
Reference
Daeffler, K. N., Galley, J. D., Sheth, R. U., Ortiz‐Velez, L. C., Bibb, C. O., & Shroyer, N. F., et al. (2017). Engineering bacterial thiosulfate and tetrathionate sensors for detecting gut inflammation. Molecular Systems Biology, 13(4), 923.