Bacterial Biosensors

General Objective: Develop a fast and specific biosensor allowing spatiotemporal analysis of stimuli

To achieve the objective mentioned above, we put forward the following hypothesis:



To validate this hypothesis we set ourselves three intermediate objectives:

First intermediate objective: Explore if and how CpxR dimerizes in live E.Coli by analyzing the various orientations of CpxR - IFP1.4 fusions.

Second intermediate objective: Analyse the temporal dynamics of the functional CpxR - IFP1.4 fusion protein by change of media.

Third intermediate objective: Analyse the spatial dynamics of the functional CpxR - IFP1.4 fusion protein by microscopy.

To validate these three objectives, we designed three experiments:

First Experiment: Build constructs with all the possible orientations between CpxR and IFP1.4 and monitor activation upon 50mM KCl stress activation

Second Experiment: Induce signal activation by stressing C-termina CpxR - split IFP1.4 constructs with 50 mM KCl, and subsequently replacing the media with a 0 mM KCl solution

Third Experiment: Analyse the spatial dynamics of the functional CpxR - IFP1.4 fusion protein by microscopy.