Team:Chalmers-Gothenburg/Parts

Interlab - iGEM Chalmers-Gothenburg 2018

BioBricks

Kozak-Aga2-Linker-GFP

In order to express GFP at the surface of our yeast cells, the yeast surface anchoring protein Alpha-Agglutin 2 (Aga2) was fused to the 3’-end of GFP. Aga2 binds with a double disulfide bond to Alpha-Agglutin 1 (Aga1) in the cell wall of the yeast. To increase chances of expression a Kozak sequence was added at the 5’-end of the Aga2 and to allow for better protein function of the individual proteins, we also added a flexible linker (GGGS)x3 in between the proteins. For more information on this BioBrick, please visit the prts registery here or our results page here

Figure 1: Plasmid design with Aga2, GFP, the spacer, and the Kozak sequence. The plasmid of origin is pSB1C3.

Secretion Factor α pro + MYR1 Myrosinase

This part was developed by our team to be used as a way to kill colon cancer cells. Myrosinase belongs to a family of enzymes involved in the plant defense. It can catalyze the hydrolysis of glucosinolates. The final product that derives from this hydrolysis has been proved to inhibit carcinogenesis and tumorigenesis in certain circumstances. The Myrosinase itself is coupled to a secretion peptide since it needs to be present in the media for it to interact with the glucosinolates. A Kozak sequence was also introduced at the begining of the protein to enhance expression. Please check out our entry in the parts registry here or our results page here for more information.

Figure 2: Plasmid design with Alphapro and Myrosinase(Myr1), the spacer and the Kozak sequence. The plasmid of origin is from pSB1C3.