Measurement

Part:BBa_K546002:Experience

Designed by: M. Hesselman   Group: iGEM11_Wageningen_UR   (2011-09-14)
Revision as of 12:10, 21 September 2011 by Youri (Talk | contribs) (→‎User Reviews)

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Applications of BBa_K546002

This part was used as an 3-oxohexanoyl-homoserine lactone (AHL) responsive element. lux pL yields low constitutive expression which results in luxR production. Complexed with luxR, AHL binds to the luxpR promoter which drives the expression of GFP.

User Reviews

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No review score entered. Youri van Nuland

Functionality of this part was tested with a Spectramax M2 spectrophotometer. This apparatus can measure fluorescence in microplates. 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 this part is the repaired BBa_F2621, the broken version is also included in the measurements.


The fluorescence measurements 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 refers to this part and clearly responds to AHL as the GFP expression rapidly increases over time. F2621-GFP refers to the broken version of F2621 (thus without the constituve luxR because R0063 was missing in this part). This broken version hardly responds to AHL.

AHL response test BBa K546002.png Team Wageningen UR 2011

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