Composite
pBAD-GFP

Part:BBa_K845000:Experience

Designed by: Grégory Hansen   Group: iGEM12_Grenoble   (2012-09-26)
Revision as of 15:25, 29 September 2012 by Greghansen (Talk | contribs) (→‎Applications of BBa_K845000)

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Applications of BBa_K845000

In order to characterize the paraBAD promoter we used a transcriptional fusion of the promoter and gfpmut2 from Alon on pUA66 in BW25113 ΔcyaA. We followed the fluorescent expression of the GFP versus different concentrations of arabinose AND cAMP over time. We made this experiment in different growth media (the different media did not affect bacterial growth):

M9 complement with 0.03% of glucose and 0.03% of acetate (1)

M9 complement with 0.1% of glycerol (2)

M9 complement with 0.1% of acetate (3)


See the protocol: http://2012.igem.org/Team:Grenoble/Biology/Protocols/AND_test

Robustness test of the AND gate:

Characterization glucose.png

Characterization acetate.png

Characterization glycerol.png

As you can see on this figure even after few hours there is no fluorescent expression if one activator is missing.


With those different experiments we demonstrated that paraBAD works as expected. It will be activated only if there is arabinose and cAMP in the medium. It appears that both cAMP concentration and time response required for a half expression of GFP depends on the medium.

For more information: http://2012.igem.org/Team:Grenoble/Biology/AND_gate

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