Part:BBa_K1408002:Experience

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Applications of BBa_K1408002

The University of Washington iGEM 2014 used this part to try and produce stable variants of a protein by producing a random mutagenesis library of a protein's sequence and inserting it into the deg2 position of our plasmid as seen here[1]. This plasmid was then transformed into PyE1 yeast which has GFP under a Gal1 promoter. We expected that putting the degron near the ends would lead to a more unstable fusion protein. This was found to be the case as seen in figure 1.

Fig1

We hypothesize that if the protein of interest is unstable the entire fusion protein is degraded and the Gal4-VP16 transcriptional activator will not activate the transcription of GFP. This should show an unstable protein producing less GFP and vice versa. However our results do not show this relationship. BbpD04,3 should be the most stable (thus producing the greatest fluorescence) followed by Bindi and then BBpD04. Further testing is underway to better characterize this system.

User Reviews

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