Team:TokyoTech/Wet Lab
Experiment Overview
Characterization
Confirmation partsMain experiment
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We modified David Karigâs Turing Pattern system so that it responds to environmental stimulus and changes according to environmental conditions. We conducted the transformation of Escherichia coli and change the spot pattern formed under blue light conditions and low temperature conditions. In the original system, IPTG was used to control the Lac promoter. However, we replaced it with the newly created hybrid promoter to achieve the goal.
In this project, we are using a gene expression system which contains Las. Therefore, we planned to use registered iGEM parts that contain LasR and GFP, RhlR and GFP for positive control. However, there was insufficient data on the gene expression intensity of BBa_K575029 which contains LasR. So in order to compensate for this, we measured the fluorescence intensity under higher LasI concentrations.
We aim to control and change the experimental system according to environmental conditions. We created two hybrid promoters by adding RhlR binding site to pYcg promoter controlled by BluR (BBa_K3259002) and pCold promoter controlled by CspA (BBa_K3259003), respectively, and used them in the project. Furthermore, as a confirmation experiment of the behaviors of these hybrid promoters, we added GFP gene downstream and confirmed their expression intensity.
