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Conclusions

In conclusion, the State 2 of our bistable i by IPTG doesn't only express the cI repressor, also it expresses the Sspb adaptor protein. This protein binds the DAS+4 tags and enhances specific degradation by the ClpXP protease. Therefore, when the State 2 is active, State 1 promoter is being repressed and the State 1 reporter is being degraded. Thus, State 2 repressor altogether

With this proteolysis system we are, as said, degrading State 1 reporter and State 2 repressor when this second State is the dominant. This will allow us both to minimize the residual GFP signal and to enhance PLac expression, as it won’t be repressed. With this mechanism, we intend to shorten switching time between states and to obtain the clearest signal difference.