Our initial plan was to both create and test microfluidic devices and the tools not only in E.coli, but also to manufacture and test it for yeast. However, as we lacked some necessary components in order to work with yeast we asked the iGEM team of the Technological University of Denmark (DTU) for assistance, which we knew after the Nordic iGEM Conference worked with yeasts. They supplied us with both a strain and required shuttle vectors, which we would have used if we had been able to perform the Yeast tests. DTU later contacted us back regarding help with their shuttle vector between E. coli and Y. lipolitica. They were trying to insert chromoproteins into the shuttle vector, but had so far been unsuccessful. As we had chromoproteins in stock from prior iGEM Uppsala teams, we moved one of the chromoproteins from the psB1A3 backbone into the shuttle vector. To simplify the assemblies we first moved the standard RFP producing insert into the previously empty insert site of the shuttle vector. Visual analysis was done in order to compare overnight production of the chromoprotein and RFP in the shuttle vector to the production of the proteins in the standard psB1A3 backbone. The comparison was done with E. coli cells transformed with the empty shuttle vector as control. These tests showed no observable difference in the production of chromoproteins in the shuttle vector compared to psB1A3.
The Aalto-Helsinki iGEM team helped us by supplying us with protocols for working with yeast.
To try their linker between spiderweb and a protein the Stockholm iGEM team asked for the UnaG protein. As UnaG requires the cofactor bilirubin to fluoresce, which can both be added and washed away, it is a good protein for characterization. We therefore assembled a plasmid with RBS and UnaG under the control of the specifically requested T7 promoter which was sent to the Stockholm team. We also supplied the Stockholm team with bilirubin as well as a guide on how to use it.
Our team also participated in this years Nordic iGEM Conference (NIC) hosted this year by the Stockholm team. NIC is a conference held during the summer to strengthen the bonds between the nordic teams and allow them to meet and exchange ideas.