Difference between revisions of "Part:BBa K2507006"
 
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<partinfo>BBa_K2507012 short</partinfo>
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<partinfo>BBa_K2507006 short</partinfo>
  
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This part consists of the fragment from iGEM 2017 kit4 2I and the plasmid backbone pSB1C3.  In combination with the <i>E. coli</i> this part can produce cjBlue  green chromoprotein.
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==Usage and Biology==
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<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.
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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>
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<partinfo>BBa_K1758300 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K2507006 SequenceAndFeatures</partinfo>
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==Characterization==
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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:
  
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https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507013
  
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<html>
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https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507014
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<h2>Performance</h2>
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<p>We use this part to test and find if specific chromoprotein can fit our requirement to replace the sfGFP as a expressor.</p>
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<<!--
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https://parts.igem.org/wiki/index.php?title=Part:BBa_K2507015
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<figure>
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<img src="https://static.igem.org/mediawiki/2015/8/84/Bielefeld-CeBiTec_arsenic_invitro_uncorrected.png" width="600px">
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<figcaption>Induction of arsenic sensor <i>in vitro</i>. For this experiment, a cell extract that already contained the arsenic repressor was used. Error bars represent the standard deviation of three biological replicates.</figcaption>
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</figure>
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-->>
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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
  
  
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<h2>References</h2>
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==Reference==
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</html>
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<p>
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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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</p>
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K2507012 parameters</partinfo>
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<partinfo>BBa_K2507006 parameters</partinfo>
 
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Latest revision as of 13:47, 1 November 2017


J23109-ttrS

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
    Illegal NheI site found at 1383
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 857
  • 1000
    COMPATIBLE 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.