Team:Paris Bettencourt/Experiments/YFP TetR diffusion experiments

From 2011.igem.org

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<h1>Testing nanotubes with the YFP concentrator system</h1>
<h1>Testing nanotubes with the YFP concentrator system</h1>
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<h2>Design overview</h2>
 
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<center><img src="https://static.igem.org/mediawiki/2011/9/93/YFP_concentration_principle.jpg"></center>
 
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<center><p>YFP:TetR/TetO array system</p></center>
 
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<p>More information on the design <a href="https://2011.igem.org/Team:Paris_Bettencourt/GFPLac_diffusion ">here</a>.</p>
 
<h2>Summary</h2>
<h2>Summary</h2>
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<h2>Design overview</h2>
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<center><img src="https://static.igem.org/mediawiki/2011/9/93/YFP_concentration_principle.jpg"></center>
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<center><p>YFP:TetR/TetO array system</p></center>
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<p>More information on the design <a href="https://2011.igem.org/Team:Paris_Bettencourt/GFPLac_diffusion ">here</a>.</p>
<h2>Emitter & receiver constructs in <i>B.subtilis</i> (receiver in plasmid)</h2>
<h2>Emitter & receiver constructs in <i>B.subtilis</i> (receiver in plasmid)</h2>

Revision as of 22:41, 28 October 2011

Team IGEM Paris 2011

Testing nanotubes with the YFP concentrator system

Summary

Results for the YFP concentrator:

  • We successfully BioBricked both the YFP:TetR (BBa_K606025) and the TetO Array (BBa_K606026) constructs and sent them to the registry
  • We characterized the YFP:TetR fusion protein both in E.coli and B.subtilis
  • We characterized the TetO array both in E.coli and B.subtilis
  • We've done E.coli to B.subtilis diffusion experiments (with negative results)


Design overview

YFP:TetR/TetO array system

More information on the design here.

Emitter & receiver constructs in B.subtilis (receiver in plasmid)

Emitter construct in E.coli - Receiver construct in B.subtilis (plasmid)

We success to integrate the TetO Array in B. subtilis with the multihost episomal pHM3 plasmid.
Experiments of E. coli (pFX234 plasmid, D. Lane strains) to B. subtilis (TetO Array, pHM3) to follow an eventual fluorescence diffusion by nanotube has been tried.

At the beginning of the movie
TetRTetO0 YFPTrans01.jpg
TetRTetO0 YFPFluo01.jpg.jpg
At the end of the movie
TetRTetO0 YFPTrans02.jpg
TetRTetO0 YFPFluo02.jpg

Movie : Experiment of mixed YFP:tetR (E. coli) & TetO Array (B. subtilis)

We cannot see spot apparitions specific of YFP concentration on TetO Array. This experiment leads us to think that there is no diffusion of the YFP:tetR fusion protein from E. coli to B. subtilis by nanotube.

Conclusions