Part:BBa_K1493002:Experience

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••••• Wageningen UR 2014

A growth experiment was set up, using M9 medium. In this experiment both the WT P. putida and P. putida containing the BBa_K1493000 BioBrick are grown in a 96-wells plate with 0, 15, 30, 45, 60 and 75µg/ml fusaric acid. Furthermore, DH5α E. coli cells (WT and BBa_K741002) were also grown in the same plate, but without fusaric acid. The transformed E. coli has a well characterized promoter with the same GFP gene downstream. By comparing the fluorescence of our fusaric acid induced promoter at different fusaric acid concentrations to this constitutive promoter, a characterization can be performed. Figure 1. The measurement is based on GFP fluorescence in P. putida at increased concentrations of fusaric acid to prove and characterize the activity of the fusaric acid induced promoter, BBa_K1493000. For comparison, the well characterized pLac promoter (BBa_K741002, uninduced by IPTG) was used to quantify the activity of this promoter at different concentrations of fusaric acid. The promoter does not respond to low concentrations up to 170nmol/ml. From 255nmol and up, the activity increases. The maximum measured activity of the promoter is 0.21 RPU at 425nmol/ml.

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