We have created a part that allows a quick engineering of a reporter gene thanks to a Golden Gate site (BBa_K2675081), another part that allows the production of a transcription factor (BBa_K2675040) and a third one that permit modulable expression of a tagged sfGFP thanks to the TF of the second part (BBa_K2675058).
As our team is leading a foundational advance project, having a discussion with public representatives appeared to be complex. To address this problem, we decided to focus on meeting with professionals to find applications to our project, and on educating the general public about synthetic biology: https://2018.igem.org/Team:Evry_Paris-Saclay/Human_Practices
GOLD MEDAL 🥇
Integrated Human Practices:
After performing researches on our own and meeting various professionals, we managed to highlight important points of our project that needed a particular attention or modifications. It helped us in the determination of the exact type of communication system we would work on and the choice of our chassis and comforted us in our engagement in the education of the public: https://2018.igem.org/Team:Evry_Paris-Saclay/Human_Practices#integrated_human_practices
Improve a Previous Part:
By adding the OmpA secretion signal to the hexapeptide SAIRGA, we managed to obtain the secretion of the peptide by E.coli, which didn't occur with the original part (BBa_K2279001), created to be used in B.subtilis. The secretion is a sine qua non condition for a functioning arbitrium system and, therefore, was needed to transfer this system in E.coli: https://2018.igem.org/Team:Evry_Paris-Saclay/Improve
Model Your Project:
We have used structural modelling to analyse in great detail the molecular interactions in the phage arbitrium system and to estimate whether the system would have been portable to E. coli. We have used the knowledge gained to guide the successful re-engineering of the aimR-aimP system into a flexible communication system for E. coli: https://2018.igem.org/Team:Evry_Paris-Saclay/Model
Demonstration of Your Work:
We have successfully transferred the ‘Arbitrium’ communication system from a Bacillus phage to E. coli. While doing so, we have engineered new “composite” promoters that convert the transcriptional activator AimR from Bacillus into a repressor in E. coli. We have also identified a putative intrinsic terminator in the pAimX(full) promoter sequence, and verified its terminator activity in E. coli. Further, we have ruled out a transcription antitermination mechanism at this terminator site. We also have promising data that indicates conditional regulation of AimR repressor activity by the small signalling peptide SAIRGA: https://2018.igem.org/Team:Evry_Paris-Saclay/Demonstrate
