Part:BBa_K1627001
Device to demethylate 3-methylxanthine
This part contains genes that allow for the demethylation of 3-methylxanthine. The part contains a synthetic operon consisting of ndmB, 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 3-methylxanthine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of 3-methylxanthine, a growth curve can be made that correlates culture growth (OD600) with concentration of 3-methylxanthine. Then if the strain is grown in a minimal media with an unknown concentration of 3-methylxanthine, the growth can be compared to the growth curve to calculate the initial concentration of 3-methylxanthine 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 3-methylxanthine. As the concentration of 3-methylxanthine 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 3-methylxanthine to the standard curve, the concentration of 3-methylxanthine 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 1207
Illegal BglII site found at 1678
Illegal XhoI site found at 2819 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 3517
Illegal AgeI site found at 1346 - 1000COMPATIBLE WITH RFC[1000]
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