Part:BBa_K3463011:Experience

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Characterization BBa_K3463011

The Figure 1 illustrates the fluorescence intensity over time for 2 growing cultures of E. coli Nissle transformed with the plasmid pUCBB PrhlR-eGFP BBa_K3463011. Cultures were performed in absence or in presence of 100µM of synthetic BHL. In addition, a culture of wild type E. coli Nissle (not transformed) was used as a negative control.

The fluorescence intensity was monitored for 15 hours. Each time point was performed hourly in triplicata. For each curve, the first measurement (T0) was set at 0 and subtracted to following ones. This experiment was repeated 3 times.

Figure 1 eGFP expression according to the BHL and over time. Two conditions with 100µM of BHL and without BHL were performed to evaluate the effect of BHL in E. coli Nissle. With and without BHL, no eGFP expression should be detected because there is no production of RhlR protein.

Averages: Without BHL=100µM BHL=Control -

As it can be seen in figure 1, all cultures have similar levels of eGFP expression over time (p-value=0,44). Indeed, we can see that the fluorescence intensity of E. coli Nissle PrhlR-eGFP increases slightly whatever the presence of BHL in the medium. In addition, the curve of negative control also increases in the same manner over time. Therefore, there is no significant expression of eGFP in E. coli Nissle PrhlR-eGFP, even with a large amount of BHL in the medium.

To conclude, we can significantly ensure that BBa_K3463011 does not allow E. coli Nissle to express downstream genes without RhlR, whatever the presence of BHL.

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