Team:ETH Zurich/labblog/20140811

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Cross talk experiments

Date : Friday, 15th August

Crosstalk Experiments

After having constructed the necessary regulator and sensor plasmids we were able to start first cross talk experiments. Therefore we transformed competent cells with different combinations of regulator and sensor plasmids. One example would be the strain siG0024 which contains the regulator plasmid piG0041 producing LuxR under a constitutive promoter and the sensor plasmid piG0051 that encodes for sfGFP. The latter is under control of a pluxR promoter, hence it will be transcribed in the presence of LuxR and Lux. To assess crosstalk and leakiness we conducted multiple experiments: after adding one of the three AHLs to the cultures we analyzed the raise in sfGFP production using a microtiterplate reader. Thus we evaluated for example the sensitivity of the piG0051 construct to different concentrations of 3OC6-HSL (LuxI product), but also its sensitivity towards 3OC12-HSL (LasI product) and C4-HSL (RhlI product). Indeed, we could observe the induction of sfGFP transcription by comparably low levels of 3OC12-HSL. Therefore crosstalk can be observed between the Las and the Lux system. All these informations we will use to design a system tailored to our plans.

ETH Zurich crosstalk luxRR12y.png


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