Difference between revisions of "Part:BBa K546002:Experience"

Revision as of 18:29, 19 September 2011

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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

UNIQbfe881ec0e9df392-partinfo-00000000-QINU

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 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.

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UNIQbfe881ec0e9df392-partinfo-00000002-QINU

@@ Line 18: / Line 18: @@
 Functionality of this part was tested with a Spectramax M2 spectrophotometer. This apparatus can measure fluorescence in microplates.
-The response to AHL was examined by applying different concentrations of AHL: 0, 250 and 1000 nM AHL in a 200 µL of overnight culture samples susepended in PBS in a 96 wells plate.
+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.