S.E.colization : Green light, Go ! Aix-Marseille University 2014 Team Advisors : Aurélie Dobric Collaborators : Dr. E. Bouveret (LISM, Marseille) Ressources : Dr. J. Dueber (Berkeley University, USA) and Dr. B. Aimeric AGAOUA Lambert ANTONI Clara BOUYX Alexia SATOUF Romain VEILLARD Vincent CASTEL Students James STURGIS Gaël CHAMBONNIER Laetitia HOUOT Sandra MICHEL-SOUZY Field (Laboratoire de Génétique et Biophysique des Plantes, Marseille) Supervisors Serine receptors (Tsr) desactivated Autophosphorylation of CheA Increased level of serine Overexpression of RelA Serine production stopped Division of bacteria Autophosphorylation of CheA inhibited Overexpression of Mesh1 Controlling cell Division : Rewiring Intracellular signaling : We control cell division using [ppGpp] modulated by RelA and Mesh1. To validate our part BBa_K1349001 (RelA coding sequence), we used a mutant unable to synthetize RelA, and growing very slowly. When we complement this strain with BBa_K1349001, bacteria normal growth is restored. Our RelA part is functionnal. To modulate expression of genes, we used phosphorylation of the regulator CusR. CusR binds to CusR box sequences in reponse to metallic stress (Cu, Ag). To validate our part BBA_K1349002 (CusR box sequence), we grew cells in the presence of Ag. As expected, 2 or 3 CusR boxes on a plasmid sequester the endogenous CusR, preventing the normal detoxication of the cells. Figure 1 : The presence of RelA gene restore a normal growth in MG1655∆relA strain Figure 2 : The presence of cusR boxes on a pLac vector affects cell growth in the presence of a sub-lethal dose of AgNO3. Modeling : Beyond the bench : « Synthetic » + « Biology »…how does it sound for the public? The Messenger has to be a trusted source (ie a scientist!) You want a balanced and credible message. Avoid sensational!!! Our goal : identify some key rules to communicate efficiently on Synthetic Biology (SB) to a broad audience. Our resources: bibliography + our own experiences “on field” (presentation to local media, sponsors, and during Be prepared to have tricky questions about safety, ethic the French Science Festival “Fête de la Science”) and responsibilities for this emerging technology. A computer simulation of the designed system shows that CusR, and ppGpp can oscillate as the system grows and serine accumulates. We implemented our simulation in Matlab, using a set of differential equations that we integrated numerically using a Runge-Kutta 4 algorithm. Figure 3 : Modeling of our biological model
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