Community Outreach

Collaborations

MIT

This summer, we were mentored by the MIT iGEM team in making and using electrocompetent cells, making chemically competent cells, Golden Gate Assembly, and organization of the team. As a longstanding, experienced team, MIT has more experience in designing experiments and getting their projects from start to finish. Initially, we were struggling to get competent cells that would transform with our parts. After speaking with MIT at the NEGEM meetup, they shared a useful protocol that, once used, yielded extremely competent cells that we used for the entirety of our project. In August, we were invited back to MIT, where we learned about chemically competent cells, Golden Gate Assembly, and general running of the team. We used the protocol and advice on chemically competent cells for our lambda red recombineering procedure, in which we had to transform cells multiple times with different plasmids to knockout two genes. This proved extremely important in our project as, without the knockouts, our MtrCAB system would have competed with other naturally-occurring pathways in the E. coli, decreasing its efficiency and effectiveness in the project. Additionally, MIT explained their work using the Universal Acceptor vector backbone with specifically-designed insert segments for easy assembly using Golden Gate. Although it was too late to use this for this year’s project as our parts were already ordered and partially assembled, we will be able to use this information, along with the toolkit they are designing this year, for future projects.

University of Toronto

We were also mentored by the UToronto team, who provided advice on how to begin with a modeling effort for our team. Brayden and Fin gave direction on what items to start with. The first suggestion was a confirmation that our plan to study the kinetics of each of the enzymes was a reasonable place to start. This study would require both an estimation and/or literature search for each of the k values for each of the enzymes involved. Next, we discussed whether the loading/diffusion of phenazine shuttles was a limiting factor for the transfer of electrons between electrode and our engineered system. This aspect of our system could potentially be modeled using a random walk model of the electrons being transferred. Both these items could be used for our future work in modeling the MES system.