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

From 2014.igem.org

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|+'''Table 3''' Crosstalk matrix for the promoter prhl ([http://parts.igem.org/Part:BBa_I14017:Experience BBa_I14017])
|+'''Table 3''' Crosstalk matrix for the promoter prhl ([http://parts.igem.org/Part:BBa_I14017:Experience BBa_I14017])
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|colspan="4" style='font-size:10pt'|In all the measurements conducted to create this matrix the promoter pLux was the basis and was induced in six different variations shown.  
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|colspan="4" style='font-size:10pt'|In this set of experiment the promoter pRhl was tested for possible crosstalk. In the top row left graph we see the the induction of pRhl by C4-HSL bound to regulator RhlR. The switching behaviour was observed at a concentration of the inducer molecule of 10<sup>3</sup> nM.
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|colspan="4" style='font-size:10pt'|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 3OC6-HSL. The light-blue curve plotted shows modeling data of pLux induced by 3OC6-HSL (Lux-AHL) binding to the corresponding LuxR regulator. This curve from the model and the dark blue data point gained in experiments were plotted as a reference in all the other graphs describing pLux. Crosstalk can be observed for the cases where the 3OC12-HSL (Las-AHL) binds the LuxR regulator.
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|colspan="4" style='font-size:10pt'|In the case of 3OC12-HSL binding the RhlR regulator and subsequently the promoter pRhl negligible crosstalk has been observed.
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Severe crosstalk was detected in the case of 3OC6-HSL binding the RhlR regulator followed by induction of pRhl. The transition occurred at a concentration of the inducer molecule of 10<sup>3</sup> nM but compared to the reference curve a lower (1000 AU) value of fluorescence was observed. The second case of crosstalk with the pRhl was detected with 3OC12-HSL binding to the corresponding LasR regulator followed by inducing the promoter pRhl. There the switching occurred at a concentration 1nM of 3OC12-HSL reaching a fluorescence of 750 AU. This is approximately 0.5 fold the value of the fluorescence shown by the reference curve in green.
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|colspan="4" style='font-size:10pt'|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 and subsequently the promoter pLux, the transition occurs at 1 nM and reaches 0.5 fold the fluorescence as pLux 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.
 
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|colspan="4" style='font-size:10pt'|The promoter pLux was not tested in combination with the RhlR regulator and the C4-HSL. 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.
 
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|style="width:200px"|[[File:ETH_Zurich_2014_qs-table_CornerRhl.png|200px|link=http://parts.igem.org/Part:BBa_I14017:Experience]]
|style="width:200px"|[[File:ETH_Zurich_2014_qs-table_CornerRhl.png|200px|link=http://parts.igem.org/Part:BBa_I14017:Experience]]

Revision as of 20:02, 17 October 2014

Table 3 Crosstalk matrix for the promoter prhl (BBa_I14017)
In this set of experiment the promoter pRhl was tested for possible crosstalk. In the top row left graph we see the the induction of pRhl by C4-HSL bound to regulator RhlR. The switching behaviour was observed at a concentration of the inducer molecule of 103 nM.
In the case of 3OC12-HSL binding the RhlR regulator and subsequently the promoter pRhl negligible crosstalk has been observed.

Severe crosstalk was detected in the case of 3OC6-HSL binding the RhlR regulator followed by induction of pRhl. The transition occurred at a concentration of the inducer molecule of 103 nM but compared to the reference curve a lower (1000 AU) value of fluorescence was observed. The second case of crosstalk with the pRhl was detected with 3OC12-HSL binding to the corresponding LasR regulator followed by inducing the promoter pRhl. There the switching occurred at a concentration 1nM of 3OC12-HSL reaching a fluorescence of 750 AU. This is approximately 0.5 fold the value of the fluorescence shown by the reference curve in green.

ETH Zurich 2014 qs-table CornerRhl.png ETH Zurich 2014 qs-table C4-HSL.png ETH Zurich 2014 qs-table 3OC6-HSL.png ETH Zurich 2014 qs-table 3OC12-HSL.png
ETH Zurich 2014 qs-table RhlR.png ETH Zurich 2014 qs-table PrhlRef.png ETH Zurich 2014 qs-table PrhlRhlRLuxAHL.png ETH Zurich 2014 qs-table PrhlRhlRLasAHL.png
ETH Zurich 2014 qs-table LuxR.png ETH Zurich 2014 qs-table PrhlLuxRRhlAHL.png ETH Zurich 2014 qs-table PrhlLuxRLuxAHL.png ETH Zurich 2014 qs-table PrhlLuxRLasAHL.png
ETH Zurich 2014 qs-table LasR.png ETH Zurich 2014 qs-table PrhlLasRRhlAHL.png ETH Zurich 2014 qs-table PrhlLasRLuxAHL.png ETH Zurich 2014 qs-table PrhlLasRLasAHL.png