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Latest revision as of 11:20, 18 September 2015
Collaborations
Aix-Marseille
For our first collaboration we aided the iGEM Team from Aix-Marseille with their project on chewing-gum.
The aim of the project was to collect answers to a questionnaire from people on the street. The questions concerned the use of chewing gum, as well as the public understanding of synthetic biology and genetic manipulation.
We asked over 30 people to fill in this questionnaire, earning us a Gold Medal for the collaboration.
Aachen
At the meetup in Germany we met the team from Aachen, who are working on "Upcycling methanol into a universal carbon source". For this project they designed synthetic pathways involving multiple different enzymes.As our project is based on the focusing of enzymatic reactions onto one scaffold, our flexosome, we had found our first application. The Aachen Team supplied us with their BioBricks for Methanoldehydrogenase II from Bacillus methanolicus (BBa_K1585210, MDH), 3-hexulose-6-phosphate synthase from Methylococcus capsulatus (BBa_K1585211, HPS)and 6-phospho-3-hexuloisomerase from Methylococcus flagellatus (Ba_K1585212, PHI). For further information refer to the Wiki from Aachen
We then modeled their enzymes into our system via Clone Manager and created the Primers that would allow us to insert them into our expression vector pBAD. Following constructs were planned:
Vector: pBAD His A (Arabinose induced expression Vektor with an N-Terminal His-Tag)
Enzyme/Dockerin Fusions: MDH & CTHE, HPS & ACEL, PHI & BCEL
Dockerins from ACEL: , A. cellulolyticus , BCEL: Bacteroides cellulosolvens, CTHE: Clostridium Thermocellum
By the end of the competition, we were able to fuse HPS to the Dockerin ACEL and PHI to the Dockerin BCEL and prove their correct insertion by sequencing. The fusion of MDH to the Dockerin CTHE was thus far not successful.