Difference between revisions of "Part:BBa K606027:Experience"

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The pictures of TetO show no YFP activity, which is normal because there is no YFP sequence in these plasmids.<br>
 
The pictures of TetO show no YFP activity, which is normal because there is no YFP sequence in these plasmids.<br>
 
The TetR-YFP construct which constitutes the transmitter part, occasionally shows gross aggregated YFP. This is not what we expected at first, but that does not prevent us to characterize the full construct.<br>
 
The TetR-YFP construct which constitutes the transmitter part, occasionally shows gross aggregated YFP. This is not what we expected at first, but that does not prevent us to characterize the full construct.<br>
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After observing the full construct's pictures, we can obviously distinguish glowing dots in some cells. They reflect the behavior we expected. Indeed, appearance of dots shows (red arrow) that the receiver (TetO array) actually links tightly to TetR-YFP which is the emitted protein. Not all the dots are highlighted with red arrows but all are fluorescence loci.<br><br>
 
  
  
 
More information on : iGEM Paris Bettencourt 2011 wiki [http://2011.igem.org/Team:Paris_Bettencourt/Experiments/YFP_TetR_diffusion]
 
More information on : iGEM Paris Bettencourt 2011 wiki [http://2011.igem.org/Team:Paris_Bettencourt/Experiments/YFP_TetR_diffusion]

Revision as of 20:03, 22 October 2011

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Please enter how you used this part and how it worked out.

Applications of BBa_K606027

User Reviews

Parts construction:
Origins of TetO Array are from pDAG479 of D. Lane (Toulouse 2 University).

Cloning plan of TetO array construction



We characterize it with arabinose-induced YFP:TetR Wild Type from pFX234 plasmid of F-X Barre and D. Lane (Kinetics of plasmid segregation, Molecular Microbiology, 2004).

Microscopy of double transformated pFX234 / Biobricked TetO Array E. Coli:
In order to do this characterization, we took pictures of different plasmids containing only TetO array (K606026); TetR + YFP (pFX234); TetO + TetR + YFP (K606026 and pFX234). In each case we made a control by non inducing the promoter with arabinose. Cells were grown at 37°C and induced at least 45min.

tetO array : 37°C
tetO / TetO array induced without arabinose on E. Coli .
tetO / TetO array induced without arabinose on E. Coli .
tetO / TetO array induced with 0,2% arabinose on E. Coli .
tetO / TetO array induced with 0,2% arabinose on E. Coli .
TetR:YFP : 37°C
tetR:YFP / TetR-YFP induced without arabinose on E. Coli .
tetR:YFP / TetR-YFP induced without arabinose on E. Coli .
tetR:YFP / TetR-YFP induced with 0,2% arabinose on E. Coli .
tetR:YFP / TetR-YFP induced with 0,2% arabinose on E. Coli .

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The pictures of TetO show no YFP activity, which is normal because there is no YFP sequence in these plasmids.
The TetR-YFP construct which constitutes the transmitter part, occasionally shows gross aggregated YFP. This is not what we expected at first, but that does not prevent us to characterize the full construct.


More information on : iGEM Paris Bettencourt 2011 wiki [http://2011.igem.org/Team:Paris_Bettencourt/Experiments/YFP_TetR_diffusion]