Coding

Part:BBa_K2507001

Designed by: Xiaoxiao Chen   Group: iGEM17_SHSBNU_China   (2017-10-01)
Revision as of 13:16, 3 October 2023 by ASHIELIW (Talk | contribs)

ThsR

Background

ThsS (BBa_K2507000) and ThsR (BBa_K2507001), both codon-optimized for E. coli, are two basic parts which belong to the two-component system from the marine bacterium Shewanella halifaxensis. ThsS is the membrane-bound sensor kinase (SK) which can sense thiosulfate outside the cell, and ThsR is the DNA-binding response regulator(RR).



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 13

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.

Jackson MR, Melideo SL, Jorns MS (2012) Human sulfide: quinone oxidoreductase catalyzes the first step in hydrogen sulfide metabolism and produces a sulfane sulfur metabolite. Biochemistry 51: 6804 – 6815

Levitt MD, Furne J, Springfield J, Suarez F, DeMaster E (1999) Detoxification of hydrogen sulfide and methanethiol in the cecal mucosa. J Clin Invest 104: 1107 – 1114

Schmidl SR, Sheth RU, Wu A, Tabor JJ (2014) Refactoring and optimization of light-switchable Escherichia coli two-component systems. ACS Synth Biol 3: 820 – 831

Vitvitsky V, Yadav PK, Kurthen A, Banerjee R (2015) Sulfide oxidation by a noncanonical pathway in red blood cells generates thiosulfate and polysulfides. J Biol Chem 290: 8310 – 8320


Bacterial thiosulfate sensors (by IGEM23_WHU-China)

Thiosulfate (S2O32-) is a promising biomarker for gut inflammation. Bacterial sensors based on ThsS/R, two-component systems, can be used to diagnose colonic inflammation (colitis) through flow cytometry analysis of colon and fecal samples by responding to thiosulfate.

In a previous study, the sensitivity and specificity of thiosulfate sensors were measured.

Figure 1:Characterization of the thiosulfate sensor ThsSR. (Kristina N-M Daeffler et al. 2017)

In another literature, the ThsS/R-mediated expression of sfGFP was optimized in response to thiosulfate at transcription to increase the induction ratio and enhance the output intensity.

Figure 2:Fluorescence output of each gene circuit. (Zhen-Ping Zou et al. 2023)

It tested the sensitivity of the biosensor to thiosulfate and the specificity by using a panel of metabolites in the gut. The result showed that the biosensor system on pWT-CS2R4 produced a detectable and especially specific signal for a minimal thiosulfate concentration (as low as 0.016 mM).

Figure 3:Thiosulfate dose response of the optimized strain E-sfGFP and EcN. (Zhen-Ping Zou et al. 2023)

Figure 4:Selectivity of ThsS/R to thiosulfate over other terminal electron acceptors. (Zhen-Ping Zou et al. 2023)

Reference

  1. Daeffler KN, Galley JD, Sheth RU, Ortiz-Velez LC, Bibb CO, Shroyer NF, Britton RA, Tabor JJ. Engineering bacterial thiosulfate and tetrathionate sensors for detecting gut inflammation. Mol Syst Biol. 2017 Apr 3;13(4):923.
  2. Zou ZP, Du Y, Fang TT, Zhou Y, Ye BC. Biomarker-responsive engineered probiotic diagnoses, records, and ameliorates inflammatory bowel disease in mice. Cell Host Microbe. 2023 Feb 8;31(2):199-212.e5.
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