Signalling

Part:BBa_K546003:Experience

Designed by: Brendan Ryback, Matthijn Hesselman   Group: iGEM11_Wageningen_UR   (2011-09-14)
Revision as of 19:06, 19 September 2011 by Youri (Talk | contribs) (→‎Applications of BBa_K546003)

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Applications of BBa_K546003

This part serves as a weak constitutive producer of luxR. luxR can form a complex with 3OC6HSL (AHL) inducing the pR promoter.

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Functionality of this part was tested with a Spectramax M2 spectrophotometer. This apparatus can measure fluorescence in microplates. The part was fused to GFP for quantitative measurements. The response to AHL was examined in duplo by applying different concentrations of AHL: 0, 250 and 1000 nM AHL in a 200 µL of overnight culture samples resuspended in PBS in a 96 wells plate. As a negative control for this experiment, part BBa_K546001 was used. This part should also respond to AHL, but without yielding a fluorescent signal.


The fluorescence measurement are depicted in the diagram underneath. The detection limit of the apparatus was 13000 RFU for this experiment, as is visible in the diagram. R0063-F2621-GFP clearly responds to AHL as the GFP expression rapidly increases over time. The F2621*-GFP part (thus without the constituve luxR) hardly responds to AHL.

AHL response test BBa K546002.png

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