Part:BBa_K1973027
miniTn7BBGm-nahR-Psal-glpF
nahR (Pseudomonas putida, see BBa_K1031610) is expressed under the constitutive Pr promoter. It encodes a regulatory protein that activates the Psal promoter (see BBa_J61051) in the presence of salicylate. glpF (BBa_K1973026) is expressed from the Psal promoter in the presence of this molecule. The miniTn7 tool allows the stable integration of this module in the chromosome of a wide range of bacteria (see https://parts.igem.org/Genome_Integration).
By measuring bacterial growth in a medium with glycerol as the only carbon source, we can explain the glycerol consumption of the different strains with and without this construct. Growth of bacteria expressing glpF (with the BBa_K1973027 part inserted in the genome) was compared to a control, bacteria with an “empty†mini-Tn7, a Tn7 with no added information. It was expected that bacteria expressing glpF in presence of an activator would reach higher growth rates than the control. We can state that modified bacteria reach a higher absorbance than control in a minimal media with glycerol 5 mM.
We then repeated the experiment adding octanoic acid 1 mM as a growth precursor. In this case, we observed lag phase was substantially reduced, and bacteria reached exponential phase when growing in glycerol during the 23 hours the experiment lasts. These results show that bacteria expressing glpF reach a higher absorbance than control in media with glycerol. we conclude that the expression of the gene encoding the glycerol transporter of the inner membrane is enough to increase consumption of this molecule, as modeling studies predicted.
Figure 1. Graphs showing bacterial growth in media with glycerol as a sole carbon source. Genetically modified bacteria reach a higher absorbance than wild type in media with salycilate.
For further information and details about the experiments, check our wiki page http://2016.igem.org/Team:UPO-Sevilla/Experiments .
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