Difference between revisions of "Part:BBa K2507013"
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==Characterization== | ==Characterization== | ||
After validate this system in laboratory <i>Escherichia coli</i> Top10 and <i>E.coli</i> Nissle 1917, this system can function as a tetrathionate sensor and reporter. | After validate this system in laboratory <i>Escherichia coli</i> Top10 and <i>E.coli</i> Nissle 1917, this system can function as a tetrathionate sensor and reporter. | ||
| − | [[File: SHSBNU 17 40a14.jpg| | + | [[File: SHSBNU 17 40a14.jpg|200px|thumb|center|alt text]] |
Figure 1. Schematic of ligand-induced signaling through TtrS/R and plasmid design of the sensor components. TtrS/R were tested under the situation BBa_K2507006 was in pSB4K5 backbone and BBa_K2507013 was in pSB1C3 backbone. We submitted the parts all to the iGEM registry in pSB1C3. | Figure 1. Schematic of ligand-induced signaling through TtrS/R and plasmid design of the sensor components. TtrS/R were tested under the situation BBa_K2507006 was in pSB4K5 backbone and BBa_K2507013 was in pSB1C3 backbone. We submitted the parts all to the iGEM registry in pSB1C3. | ||
[[File: SHSBNU 17 40a23.jpg|200px|thumb|left|Figure2]] | [[File: SHSBNU 17 40a23.jpg|200px|thumb|left|Figure2]] | ||
Revision as of 10:41, 1 November 2017
J23109-ttrR-PttrB185-sfGFP
Usage and Biology
E.coli codon optimized TtrS(BBa_K2507002) and TtrR(BBa_K2507003) are two basic parts which belong to the two-component system from marine 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 when TtrR is phosphorylated by TtrS after TtrS sensing tetrathionate. Winter et.al have shown that reactive oxygen species (ROS) produced by the host during inflammation convert thiosulfate to tetrathionate, which this pathogen consumes to establish a startpoint for infection (Winter et al, 2010).Thus, tetrathionate may correlate with pro-inflammation conditions and can be used as gut inflammation sensor.
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
After validate this system in laboratory Escherichia coli Top10 and E.coli Nissle 1917, this system can function as a tetrathionate sensor and reporter.
Figure 1. Schematic of ligand-induced signaling through TtrS/R and plasmid design of the sensor components. TtrS/R were tested under the situation BBa_K2507006 was in pSB4K5 backbone and BBa_K2507013 was in pSB1C3 backbone. We submitted the parts all to the iGEM registry in pSB1C3.
Figure3. We cultivated E.coli Nissle 1917 overnight under aerobic or anaerobic condition. Ths/R-sfGFP seems act better under anaerobic condition. While TtrS/R-sfGFP acted much worse than ThsS/R. So in TtrS/R system, we replaced gfp gene by vioABDE(BBa_...17). It worked better.
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.