To measure our split-ribozyme we used GFP as a reporter. We assembled a plasmid with an inducible GFP construct as control to compare different levels of translation. The part BBa_K2406020 is a T7 promoter combined with the LacO promoter to make it IPTG inducible. Downstream of the promoter is the strong binding site BBa_B0034 with the standard GFP BBa_E0040 and the BBa_B0015 Terminator. We first measured the construct without inducing it with IPTG and found that it had a high fluorescence. So we could not use it for different levels of GFP as a comparison to our split ribozyme results. The promoter is too leaky. We contributed these measurements to the corresponding part pages in the registry. In figure 1 you can see a plate containing different bacterial colonies with different constructs. Very clearly you can see our non-induced GFP controls fluoresce in the UV light.
To help other teams use the split ribozymes we developed a software. The input for the software is simply your target sequence and the sequence of a reporter of your choice. You will then get the complete sequence for ordering with the biobrick prefix and suffix for easy assembly into your backbones.
Outside of the lab we wanted to contribute to the iGEM community in a special way. iGEM as a community is all about working together and networking. How could that be better achieved than in a meetup? The last two years suffered under harsh covid security measures and no meetups were possible. So it is even more important to start the vibrant culture of iGEM meetups again! We worked together with iGEM Toulouse and Münster to create a How-to-Meetup guide that will help teams in the next years organize their own meetups to help iGEM become the great interconnected community that it was before covid.
