Part:BBa_K2771030:Experience

USP-Brazil Characterization

This part has been tested alongside with six quorum sensing promoters, pLux (BBa_K2771021), pLas (BBa_K2771022), pRhl(BBa_K2771023), pCin(BBa_K2771024), pTra (BBa_K2771025) and pRpa (BBa_K2771026). Assay in 2mL LB in 24-well plate, shaking at 180rpm and 37ºC, 2.5 mM arabinose, taking samples every 3hrs. Before measuring each sample, we centrifuged the cells and resuspended the sample in H2O, to eliminate LB autofluorescence. Note that Y axis is in ratiometric activity, because reporter is measured as a ratio of YFP/CFP fluorescence. For excitation-emission wavelenghts, we used 430-480 for CFP and 500-530 for YFP:

Figures 1 and 2: Labels refer as Sender plasmid:Receiver plasmid, combining BBa_K2771030 with different quorum sensing promoters

We also did an assay comparing activity when induced by arabinose and when induced directly by 10^-4mM HSL. Auto-inducing showed slower response, but much higher threshold (levels and speeds slower than in the last graphs due to growing in M9 media instead of LB:

When testing the promoter of tha Las system, however, it only showed activity similar to what's seen with LB with the synthetic HSL induction, what leads us to believe that the culture couldn't reach a concentration of 10^-4mM HSL with the synthase in M9 medium:

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