Team:Gothenburg/Parts/Part Submission

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Part Submisson

Since our whole generation counter is based on cell cycle specific signals for activation of genes and degradation of proteins, we thought it would be beneficial to add one of the specific degradation signals for yeast into the registry. S. cerevisiae was established as one of the most researched on and industrially used organisms. For this reason, we planned to submit the degradation-tag we fused to the fluorescence proteins to make sure they are degraded and do not leak into the next cycle.


Figure 1. Picture of the part in SnapGene.

This degradation-tag, in short D-tag, is a short part of the cyclin Clb3. It is the sequence for the amino acids of the N-terminal until and including a destruction box, a nine amino acid recognition site with the sequence R-x-x-L-x-x-x-x-N. This sequence is necessary but not sufficient for a specific ubiqitin-mediated degradation and to be found in most of the B-cyclins of yeast.
Cyclins bind to cell cycle specific kinases and mediate thereby the progress of the different steps of the cell cycle. The six B-cyclins in yeast (Clb1-6) are responsible for the shift into the "M-phase bla". With increasing digit the degradation occours later in the cell cycle. This goes hand in hand with the position of the destruction box in the protein, which distance to the N-terminal increases with the digit as well.
To have a suitable time point of degradation for our counter we decided on using the sequence of Clb3.

Summary

Our part submission is a 165 bp long N-terminal sequence of the S.cerevisiae protein Clb3. It is a cell cycle dependend degradation signal for proteins.

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