Part:BBa_K1627005
Device to demethylate theobromine
This part contains genes that allow for the demethylation of theobromine. The part contains a synthetic operon consisting of ndmB, ndmC, and ndmD from Pseudomonas putida CBB5 and gst9 from Janthinobacterium sp. Marseille under the control of a constitutive promoter, J23110.
This part can be used in a ∆guaB strain of E. coli to measure the amount of theobromine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of theobromine, a growth curve can be made that correlates culture growth with concentration of theobromine. Then if the strain is grown in a minimal media with an unknown concentration of theobromine, the growth can be compared to the growth curve to calculate the initial concentration of theobromine in the media.
See the [http://2015.igem.org/Team:Austin_UTexas/Project/Caffeine 2015 Austin UTexas wiki].
Usage and Biology
This device was grown in the ∆guaB strain in minimal media cultures with different concentrations of theobromine. As the concentration of theobromine increases (up to the point of saturation, after which the strain won't grow further), the culture density increases linearly. By comparing growth from unknown concentrations of theobromine to the standard curve, the concentration of theobromine can be measured.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 644
Illegal NheI site found at 839 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1595
Illegal BglII site found at 2091
Illegal BglII site found at 2562
Illegal XhoI site found at 3703 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 4401
Illegal AgeI site found at 2230 - 1000COMPATIBLE WITH RFC[1000]
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