Composite

Part:BBa_K769022:Experience

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

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Applications of BBa_K769022

Evaluation

Transformed E coli. TOP10 was precultured into LB medium and incubated at 37℃. After incubating for 12 hour, precultured solution was diluted with LB medium to the point that OD660 became 0.6, then applied into 96well microtiter plate. After adding arabinose(0.25% f.c.), the plate was shaken at 20 ℃ to express the lux proteins. The spectrum of the light and intensity was measured by plate reader once in an hour.

Result

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.

Lux+GFP.jpg



We observed the maximal absorption of Lux+GFP at 512 nm (Figure.b). We thought that the shift of emitting light wavelength was caused by BRET. After 1 hour culture, peak could not be observed at 512 nm, and we performed the culture in dark condition. So it is possible to occur that GFP absorbes the lux luminescence and emits fluorescence.

User Reviews

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