With our projects united in their focus on outer membrane vesicles (OMVs), Northwestern University and UNSW came together for what felt like a very natural collaboration. Skype calls and emails enabled our two teams to overcome the distance barrier, and share critical information and advice that aided the progression of our two projects.
Northwestern University began their project with the use of a ready-prepared TolA knock-out strain of E. coli that also overexpressed TolR. This strain was given to them by a laboratory contact who had already demonstrated demonstrated OMV overproduction in this strain. For our team, this information validated our choice to include a TolA knock-out strain, and TolR overexpression strain in our library of E. coli for comparison of OMV production alongside our other strains. By including these two strains in our library and comparing their OMV production capabilities against other strains, our results can help validate the results of Northwestern University.
Importantly, throughout the iGEM competition both of our teams openly shared protocols for OMV purification. These protocols were essential for the success of both our teams, and by actively sharing feedback, troubleshooting and advice on the operation of these experiments, our results were largely furthered. For example, our team suggested alternatives to ultracentrifugation, since access was an issue for Northwestern University, and we both shared outcomes achieved with the use of different filter sizes. Furthermore, both of our teams were also testing periplasmic localisation of fluorescent proteins, and thus this was another area of protocol and results sharing.
This was a far-reaching collaboration, that reached the core of both of our projects, and we thank Northwestern University for their extensive communication with our team throughout the competition.
Please visit the experiment page to see details of our protocols, including the ones developed with the Northwestern University iGEM team.
Unlike our collaboration with Northwestern University, our partnership with the University of Sydney iGEM team was not limited by geographical distance. Thus, we made the most of our ability to share wet-lab materials and perform experiments in order to validate each other’s expression systems.
To help us, USYD heard our woes about being unable to express our parts, and stepped up by cloning BBa_2022000 and BBa_2022010 into their pET expression vector. Interestingly, upon using a Western Blot to detect expression, we found that our although their cells expressed our RFP-fusion part, they weren't turning red, which was the first hint that there was an issue with our part...
To help USYD, we performed a Western Blot on induced cells (supposedly) expressing EtnR1 and EtnR2, the two proteins that underpin their project. We determined that they were in fact expressing, and were delighted to let them know that!
Outside of the lab, we attended a STEM presentation together with the USYD iGEM team, where we opened the eyes of school students to the possibilities of having a career in the STEM field.
We also attended an evening event hosted by the University of Sydney, for Australian iGEM teams to practice their presentations before the Giant Jamboree.
The Paris-Bettencourt iGEM team reached out to us early on in the iGEM season, due to our team’s proximity to a wine-growing region. Paris Betterncourt’s project focused on wine-fermenting bacteria, and thus required biological samples from wineries (such as soil, branches, leaves and grapes).
Our team made the road trip to a wine valley in upper NSW, and collected some samples for Paris Bettencourt. We posted these to Paris Bettencourt, who graciously received them. We hope this small contribution on our part helped their project as much as it could!