We are proud to improve roGFP(BBa_K1093013) of Team iGEM13_Warsaw with our part (BBa_K2271024). In order to use the redox sensing in vivo sensor we optimized it for our application in yeast. Carboxyl terminal fusion of the peroxisomal targeting signal 1 (PTS1) resulted in small areas with high fluorescence intensity inside the cell - proven to be peroxisomes. In contrast, cytosolic fluorescence was nearly silenced. The Pex5 receptor recognizes the signaling tag PTS1 and ensures the import into the peroxisome, which is shown in the following figure.

Right: localisation of roGFP2 into the peroxisomes, Left: wild type control

Expression of our level2 plasmids containing both, a sensor and peroxin13 mRuby, a peroxisomal marker, showed co-localization which leaded to yellow spots in merged channels et al., (2009) .

Right: peroxin13-mruby and cytosolic expressed roGFP2, Left: PTS1 tagged roGFP2 co-localized with peroxin13-mruby

We successfully validated roGFP2 and conducted calibration and in vivo measurements.