Team:ETH Zurich/expresults/qs/tab-plux


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Table 1 Crosstalk matrix for the promoter pLux (BBa_R0062)

In all the measurements conducted to create this matrix the promoter pLux (ref registry same as above) was the basis and was induced in six different variations shown. The dark blue points in the graph top left show the activation of gene expression when pLux is induced by 3OC6-HSL (Lux-AHL) binding to the corresponding LuxR regulator. The observed transition occurs at a concentration of approximately 1 nM of 3OC6-HSL. The light-blue curve plotted shows modeling data of pLux induced by 3OC6-HSL (Lux-AHL) binding to the corresponding LuxR regulator (link to registry). This curve from the model and the dark blue data points obtained from experiments were plotted as a reference in all the other graphs describing pLux(ref registry). Crosstalk can be observed for the cases where the 3OC12-HSL (Las-AHL) binds the LuxR (link to registry)regulator. Additionally for 3OC12-HSL binding to its corresponding regulator LasR and then binding to the pLux as seen in the middle of the top row and center of the matrix. For the case of Las-AHL binding the LasR (ref)and subsequently the promoter pLux(ref), the transition occurs at 1 nM and reaches 0.5 fold the fluorescence as pLux (ref) induced by 3OC6-HSL binding LuxR. In the case of 3OC12-HSL binding LuxR and inducing the promoter pLux, the transition is observed at approximately 100 nM and severe crosstalk is observed, meaning that the ON-OFF-ratio is not significantly different from the reference curve.

Observation of C4-HSL has shown, that there is no significant crosstalk with the LuxR and LasR regulators binding C4-HSL and subsequently to pLux. This is indicated on top right and middle right graphs. However, RhlR induced with its corresponding inducer (C4-HSL) binds to pLux (ref) and activates expression of GFP at about 100 nM.

ETH Zurich 2014 qs-table CornerLux.png ETH Zurich 2014 qs-table 3OC6-HSL.png ETH Zurich 2014 qs-table 3OC12-HSL.png ETH Zurich 2014 qs-table C4-HSL.png
ETH Zurich 2014 qs-table LuxR.png ETH Zurich 2014 qs-table PluxRef.png ETH Zurich 2014 qs-table PluxLuxRLasAHL.png ETH Zurich 2014 qs-table PluxLuxRRhlAHL.png
ETH Zurich 2014 qs-table LasR.png ETH Zurich 2014 qs-table PluxLasRLuxAHL.png ETH Zurich 2014 qs-table PluxLasRLasAHL.png ETH Zurich 2014 qs-table PluxLasRRhlAHL.png
ETH Zurich 2014 qs-table RhlR.png ETH Zurich 2014 qs-table PluxRhlRLuxAHL.png ETH Zurich 2014 qs-table PluxRhlRLasAHL.png ETH Zurich 2014 qs-table PluxRhlRRhlAHL.png