Composite

Part:BBa_K769022

Designed by: Tomomi YOKOYAMA   Group: iGEM12_Tokyo-NoKoGen   (2012-09-19)

lux operon(K769011) + GFP(I13522)

lux operon(K769011) + GFP(I13522)
Safety.png


Why we use GFP ?

The wavelength of bacterial luciferase from Lux operon of Vibrio fischeri is close to the wavelength that excites the GFP.
We considered that BRET might occur between the bacterial luciferase and the GFP, so we designed the construct of Lux operon with GFP.

BRET

Bacterial luciferase from Lux operon emits light by catalyzing oxidization of luciferin. Oxidized luciferin, oluciferine, has high energy. When the oxiluciferin transits from high energy state to low energy state, light is emitted. On the other hand, fluorescence substance, such as GFP, absorbs the light and becomes high energy state. And when the fluorescence substance transits from high energy state to low energy state, light is emitted.

800px-Bret3.png

If the luciferase exists near the fluorescence substance, Bioluminescence Resonance Energy Transfer (BRET) occurs. BRET is a phenomenon that can be observed when a fluorescence substance is excited by obtaining energy from the bioluminescence. This only happens when the wavelength of bioluminescence is the same as the wavelength that excites the fluorescence substance.

Luxphoto.jpg
The flask on the very left is the strain that is expressing both lux operon and GFP. As you compare the strains, the color shifted a little bit towards green.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal NheI site found at 1680
    Illegal NheI site found at 5955
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 6417
    Illegal BamHI site found at 1144
    Illegal XhoI site found at 3767
    Illegal XhoI site found at 4275
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1965
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1859
    Illegal BsaI.rc site found at 2541
    Illegal BsaI.rc site found at 9263
    Illegal SapI site found at 961


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