Team:Paris Bettencourt/Experiments/YFP TetR diffusion experiments

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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
Mix of E. coli (YFP:tetR) and B. subtilis (TetO Array) - Trans image
Mix of E. coli (YFP:tetR) and B. subtilis (TetO Array) - Fluo image
At the end of the movie
Mix of E. coli (YFP:tetR) and B. subtilis (TetO Array) - Trans image
Mix of E. coli (YFP:tetR) and B. subtilis (TetO Array) - Fluo image

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