Team:NTNU Trondheim/Parts

Our composite parts comprise a double terminator sequence (BBa_B0014), one of four promotors that we investigated (BBa_K1840000 - BBa_K1840003) and the reporter gene mCherry, codon optimized for expression in Pseudomonas putida (BBa_K1840004).

Figure 1 shows as an example how the devices are composited, with edd as promotor.

Figure 1. Composition of devices with edd as promotor.

The terminator sequence is necessary, since the next gene upstream is encoded on the opposite strand. Thus, the influence of the promotor to other genes than the reporter gene can be prevented.

As stated in the project description, the promotors are regulated by derivatives of glucose. 2-Ketogluconateprevents the repression of the kgu and gad promotors, 2-Keto-3-deoxy-6-phospho-gluconate the repression of the edd and zwf promotors. Whereas 2-Ketogluconate is build up relatively early in the compartmentalized glucose metabolism of Pseudomonas putida, 2-Keto-3-deoxy-6-phospho-gluconate is one of the last intermediate before the Kreb-cycle. However, 2-Keto-3-deoxy-6-phospho-gluconate is certainly produced when glucose is taken up by P. putida. In contrary, 2-Ketogluconate is only produced in one of three possible pathways that glucose can use to pass the inner membrane.