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Applications of BBa_I14017

User Reviews

UNIQ35df54c9240041ad-partinfo-00000000-QINU

••••

ETH Zurich 2014

Characterization of two-order crosstalk on the promoter

Background information

System considered

Modeling crosstalk

First-order crosstalk

First Level crosstalk: RhlR binds to different HSL and activates the promoter

Second Level crosstalk: other regulatory proteins, like LuxR, bind to their natural HSL substrate and activates the promoter

Second order crosstalk: Combination of both cross-talk levels

Other regulatory proteins, like LuxR, bind to different HSL and activates the promoter.

Results

**Table 3** Crosstalk matrix for the promoter prhl (BBa_I14017)
In this set of experiments the promoter pRhl was tested for potential crosstalk. In the top left position we observe the induction of pRhl by C4-HSL bound to the regulator RhlR. The switching behaviour was observed at a C4-HSL concentration of 1 μM. In the case of 3OC12-HSL binding the RhlR regulator and subsequently the promoter pRhlinsignificant crosstalk has been observed. Severe crosstalk was observed 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 1 μM but compared to the reference curve a lower value of fluorescence per OD was observed (1000 a.u.). Another case of crosstalk with the pRhl was detected with 3OC12-HSL binding to the corresponding LasR regulator followed by inducing the promoter pRhl. Here switching occurred at a concentration 1 nM of 3OC12-HSL and reached fluorescence per OD of 750 a.u.. This is approximately 0.5 fold the value of the fluorescence per OD shown by the reference curve indicated in green.

Antiquity

This review comes from the old result system and indicates that this part did not work in some test.

UNIQ35df54c9240041ad-partinfo-00000003-QINU

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-In this set of experiments the promoter [https://parts.igem.org/Part:BBa_R0071 pRhl] was tested for potential crosstalk. In the top left position we observe the induction of [https://parts.igem.org/Part:BBa_R0071 pRhl] by C4-HSL bound to the [https://parts.igem.org/Part:BBa_C0171 regulator RhlR]. The switching behaviour was observed at a C4-HSL concentration of 1 μM.
+In this set of experiments the promoter [https://parts.igem.org/Part:BBa_I14017 pRhl] was tested for potential crosstalk. In the top left position we observe the induction of [https://parts.igem.org/Part:BBa_I14017 pRhl] by C4-HSL bound to the [https://parts.igem.org/Part:BBa_C0171 regulator RhlR]. The switching behaviour was observed at a C4-HSL concentration of 1 μM.
-In the case of 3OC12-HSL binding the [https://parts.igem.org/Part:BBa_C0171 RhlR regulator] and subsequently the [https://parts.igem.org/Part:BBa_R0071 promoter pRhl]insignificant crosstalk has been observed.
+In the case of 3OC12-HSL binding the [https://parts.igem.org/Part:BBa_C0171 RhlR regulator] and subsequently the [https://parts.igem.org/Part:BBa_I14017 promoter pRhl]insignificant crosstalk has been observed.
-Severe crosstalk was observed in the case of 3OC6-HSL binding the [https://parts.igem.org/Part:BBa_C0171 RhlR regulator] followed by induction of [https://parts.igem.org/Part:BBa_R0071 pRhl]. The transition occurred at a concentration of the inducer molecule of 1 μM but compared to the reference curve a lower value of fluorescence per OD was observed (1000 a.u.). Another case of crosstalk with the [https://parts.igem.org/Part:BBa_R0071 pRhl] was detected with 3OC12-HSL binding to the corresponding [https://parts.igem.org/Part:BBa_C0179 LasR regulator] followed by inducing the promoter [https://parts.igem.org/Part:BBa_R0071 pRhl]. Here switching occurred at a concentration 1 nM of 3OC12-HSL and reached fluorescence per OD of 750 a.u.. This is approximately 0.5 fold the value of the fluorescence per OD shown by the reference curve indicated in green.
+Severe crosstalk was observed in the case of 3OC6-HSL binding the [https://parts.igem.org/Part:BBa_C0171 RhlR regulator] followed by induction of [https://parts.igem.org/Part:BBa_R0071 pRhl]. The transition occurred at a concentration of the inducer molecule of 1 μM but compared to the reference curve a lower value of fluorescence per OD was observed (1000 a.u.). Another case of crosstalk with the [https://parts.igem.org/Part:BBa_I14017 pRhl] was detected with 3OC12-HSL binding to the corresponding [https://parts.igem.org/Part:BBa_C0179 LasR regulator] followed by inducing the promoter [https://parts.igem.org/Part:BBa_I14017 pRhl]. Here switching occurred at a concentration 1 nM of 3OC12-HSL and reached fluorescence per OD of 750 a.u.. This is approximately 0.5 fold the value of the fluorescence per OD shown by the reference curve indicated in green.
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