Difference between revisions of "Part:BBa K2507013"

Revision as of 11:57, 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 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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 11
Illegal NheI site found at 34
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 318
1000
COMPATIBLE 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.

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.

Figure2

Figure3

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.

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 ==Usage and Biology==
-<i>E.coli</i> codon optimized TtrS(BBa_K2507002) and TtrR(BBa_K2507003) are two basic parts which belong to the two-component system from marine <i>Shewanella baltica</i>. 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.
+<i>E. coli</i>-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 <i>Shewanella baltica.</i> 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 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.
+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.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===