Part:BBa_K1973009
miniTn7BB-Gm-nahR-Psal-nasF-lapG
nahR (Pseudomonas putida, see BBa_K1031610) is expressed under the constitutive Pr promoter. It encodes a regulatory protein that activates the Psal promoter in the presence of salicylate. lapG (see BBa_K1973000) is expressed from the Psal promoter (see BBa_J61051) in the presence of this molecule. nasF (see BBa_K1973001) is an attenuator of transcription elongation (terminator) from Klebsiella pneumoniae that decreases the basal expression from the Psal promoter. 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).
Regarding the biofilm growth, KT2442 lapG- (mutant with no LapG activity) produces 2-3 times more biofilm than the wild type and does not disperse the biofilm due to the fact that KT2442 lapG- is a biofilm superproducer. The complementation of lapG in the lapG mutant under the nahR-Psal induced expression system causes a delay in biofilm formation, a reduction of the maximum amount of biofilm and the biofilm dispersion. The expression of lapG in the wild type does not cause a big change in the behavior of the biofilm development, it only produces a little reduction of the amount of biofilm.
Therefore, we can conclude that the expression of lapG under the nahR-Psal system allows the mutant bacterium to disperse biofilm again, when the inductor (salicylate) is added to the media.
Figure 1. Serial dilution-based growth curves of the expression system nahR-Psal-lapG (20 hours). The image represents the growth curves of the strains wild type in LB (A), wild type in LB+salicylate 2 mM (B), lapG- in LB (C) and lapG- in LB+salicylate 2mM (D). We assay the strains wild type Tn7 Ø and lapG- Tn7 Ø concurrently to compare the induction effect of the expression system. Each blot represent the average of 24 data.
For further information, check our wiki page http://2016.igem.org/Team:UPO-Sevilla/Experiments .
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