Gameplay overview

We had problems with the leakiness of our system and the missing promoter library in Lyon, but solve those for Boston.
After finding out the analytical solution (see here of the solution) needed for the second promoter detecting two mines around, we decided to do partial sensitivity recovery of the first P_LuxR variant (G1) (see here for the library). This worked very well and we isolated promoters with different EC₅₀. Then thanks to the model prediction we could easily select one of those 7 promoters which fits well in our system (single mutant 1 , SM1).
Meanwhile the leakiness could be reduced by introducing a positive feedback loop (see here the optimization of the circuit) and our two problems were finally solved.

The GFP-RFP gameplay

First of all we set-up a simple version of the game using GFP and RFP as reporters for 1 and 2 mines respectively, using the newly isolated and predicted P_LuxR (SM1) as promoter in the P_Lac-LuxR-P_LuxR [http://parts.igem.org/Part:BBa_R0062 BBa_J09855] construct with RFP as reporter and the native [http://parts.igem.org/Part:BBa_R0062 BBa_R0062] in the P_Lac-LuxR-P_LuxR [http://parts.igem.org/Part:BBa_J09855 BBa_J09855] construct with the GFP as reporter. Unfortenalty the experiment on the day of the wiki freeze failed and the data will be provided at the presentation hopefully.

The hydrolase gameplay

As mentioned above we only managed to solve our major problems between Lyon and Boston. Due to time issues we were not able to finish the cloning for the wiki freeze but we will hopefully finish it this week and therefor show it at the presentation with the according video.

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