Part:BBa_K1627002

Device to demethylate 7-methylxanthine

This part contains genes that allow for the demethylation of 7-methylxanthine. The part contains a synthetic operon consisting of ndmC, 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 7-methylxanthine in a solution. By growing the ∆guaB strain with this device in a minimal media with different known concentrations of 7-methylxanthine, a growth curve can be made that correlates culture growth with concentration of 7-methylxanthine. Then if the strain is grown in a minimal media with an unknown concentration of 7-methylxanthine, the growth can be compared to the growth curve to calculate the initial concentration of 7-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 7-methylxanthine. As the concentration of 7-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 7-methylxanthine to the standard curve, the concentration of 7-methylxanthine can be measured.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 498
  Illegal BglII site found at 994
  Illegal BglII site found at 1465
  Illegal XhoI site found at 2606
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3304
  Illegal AgeI site found at 1133
  
• 1000
  COMPATIBLE WITH RFC[1000]