Difference between revisions of "Part:BBa K2507007"
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<partinfo>BBa_K2507007 short</partinfo> | <partinfo>BBa_K2507007 short</partinfo> | ||
| + | ==Usage and Biology== | ||
| + | <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 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. | |
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===Usage and Biology=== | ===Usage and Biology=== | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2507007 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2507007 SequenceAndFeatures</partinfo> | ||
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| + | ==Characterization== | ||
| + | After validating the system in the laboratory strains <i>Escherichia coli</i> Top10 and <i>E. coli</i> Nissle 1917, we confirmed that the system indeed works as a thiosulfate sensor, as intended. By linking <i>thsR</i> with <i>sfgfp</i> (BBa_K2507008), chromoprotein genes (BBa_K2507009, BBa_K2507010, BBa_K2507011) or the violacein producing operon vioABDE (BBa_K2507012), this system can respond to thiosulfate by producing a signal visible to the naked eye, either under normal or UV light, such as sfGFP, chromoproteins (spisPink-pink chromoprotein, gfasPurple-purple chromoprotein, amilCP-blue chromoprotein) or a dark-green small-molecule pigment (protoviolaceinic acid). | ||
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| + | Check the links below to learn about the details of the results: | ||
| + | |||
| + | https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507013 | ||
| + | |||
| + | https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507014 | ||
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| + | https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507015 | ||
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| + | https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507016 | ||
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| + | https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507017 | ||
| + | ==Reference== | ||
| + | <p> | ||
| + | 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. | ||
| + | </p> | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2507007 parameters</partinfo> | <partinfo>BBa_K2507007 parameters</partinfo> | ||
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Latest revision as of 13:47, 1 November 2017
J23109-ttrR
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
After validating the system in the laboratory strains Escherichia coli Top10 and E. coli Nissle 1917, we confirmed that the system indeed works as a thiosulfate sensor, as intended. By linking thsR with sfgfp (BBa_K2507008), chromoprotein genes (BBa_K2507009, BBa_K2507010, BBa_K2507011) or the violacein producing operon vioABDE (BBa_K2507012), this system can respond to thiosulfate by producing a signal visible to the naked eye, either under normal or UV light, such as sfGFP, chromoproteins (spisPink-pink chromoprotein, gfasPurple-purple chromoprotein, amilCP-blue chromoprotein) or a dark-green small-molecule pigment (protoviolaceinic acid).
Check the links below to learn about the details of the results:
https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507013
https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507014
https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507015
https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507016
https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507017
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.