Team:Paris Bettencourt/Experiments/tRNA diffusion

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Team IGEM Paris 2011

Results

We managed to characterize the tRNA biobrick by taking pictures under fluorescence lamp and microscopy of the strains described in the following.

GFP amber

One of our strains contains only the plasmid with the GFP amber under the control of the constitutive promotor Pveg, without the amber suppressor tRNA. In this situation cells do not glow. This obervation constitutes a reliable negative control.

GFP amber with tRNA amber

The second strain contains the full construct and the production of tRNA is tuned by PHyperspank which is activated by IPTG. In order to make a negative control we plated them without inducing with IPTG. In this situation cells glows with a lower intensity than with the previous construct. We can also notice that some of the cells strongly leak.

GFP amber with tRNA amber induced by IPTG

The last result is given by testing the full construct with IPTG induced cells. Each cell glows with a very weak variability and the luminance is significant.

Microscopy

GFP amber / E-coli at 37°C
E-Coli at 37°C (trans image)
E-Coli at 37°C (gfp image)
E-Coli at 37°C (trans image)
E-Coli at 37°C (gfp image)
GFP amber+tRNA amber-IPTG/ E-coli at 37°C
E-Coli at 37°C (trans image)
E-Coli at 37°C (gfp image)
E-Coli at 37°C (trans image)
E-Coli at 37°C (gfp image)
GFP amber + tRNA amber + IPTG/ E-coli at 37°C
E-Coli at 37°C (trans image)
E-Coli at 37°C (gfp image)
E-Coli at 37°C (trans image)
E-Coli at 37°C (gfp image)


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