Part:BBa_K606026:Experience
Characterisation of BBa_K606026 - TetO Array
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).
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.
| 350px|thumb|center|tetR:YFP-tetO / full construct inducted with 0,2% arabinose on E. Coli . |
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.
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 that the receiver (tetO array) actually links tightly to tetR-YFP which is the emitted protein.
More information on : iGEM Paris Bettencourt 2011 wiki [http://2011.igem.org/Team:Paris_Bettencourt/Experiments/YFP_TetR_diffusion]
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