Difference between revisions of "Team:Bielefeld-CeBiTec/Results/toolbox"

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We designed the concept of a light influenced lycopene production as basis for the establishment of a photo switchable enzyme activity. We introduced an amber-codon in the gene for crtI in the lycopene pathway and thus stopped the lycopene production. We then showed that the lycopene production can be partially retained, by cotransforming with an aaRS and cultivation without the matching ncAA.
 
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We designed an aminoacyl-tRNA-syntethase (aaRS) for the incorporation of 2-nitrophenylalanine (2-NPA) based on an experiment from Liu et al. (2009) and cloned it into both the pSB1C3 to send it to the head quarter and in pSB3K5 for further characterization. We also designed a fusion protein of GFP and streptavidin with a linker containing an amber codon and showed that only the GFP unit is expressed under normal conditions.The whole fusion protein is produced if cotransformed with the 2-NPA-RS.
 
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Revision as of 10:38, 4 October 2017

Toolbox Overview

Analysing

Photoswichting

We designed the concept of a light influenced lycopene production as basis for the establishment of a photo switchable enzyme activity. We introduced an amber-codon in the gene for crtI in the lycopene pathway and thus stopped the lycopene production. We then showed that the lycopene production can be partially retained, by cotransforming with an aaRS and cultivation without the matching ncAA.

Labeling

Photolysis

We designed an aminoacyl-tRNA-syntethase (aaRS) for the incorporation of 2-nitrophenylalanine (2-NPA) based on an experiment from Liu et al. (2009) and cloned it into both the pSB1C3 to send it to the head quarter and in pSB3K5 for further characterization. We also designed a fusion protein of GFP and streptavidin with a linker containing an amber codon and showed that only the GFP unit is expressed under normal conditions.The whole fusion protein is produced if cotransformed with the 2-NPA-RS.