Part:BBa_K953007

Agrobacterium tumefaciens Bacteriophytochrome with T7 + Heme Oxygenase (E. coli Codon Optimised)

This composite couples Agrobacterium tumefaciens bacteriophytochrome with T7 promoter (Part BBa_K953003) and Synechocystis Heme Oxygenase (Part BBa_K953004), each with an independent RBS and has been codon optimised for expression in E. coli. This results in a light-dependent biological switch that, in the presence of red light, will produce a visible colour change. Note that this part differs from Part BBa_K953005 in that the two parts in this composite are switched around.

This mechanism works by heme oxygenase, which will oxidise heme into biliverdin and can then complex with bacteriophytochrome. This complex can then absorb red light (620-750 nm) to excite the bacteriophytochrome and result in a phenotypic change from blue to green. This can be reversed by far-red light (700-800 nm) or will revert from green to blue over time. As E. coli does not produce biliverdin, heme oxygenase must be coupled with a bacteriophytochrome to activate the oxidation of heme to produce biliverdin.

The results and characterisation of our heme oxygenase/bacteriophtyochrome switch can be found here and you can also check out our entire project be clicking here.

Visualisation of digested composite biobricks. The upper band (black box) is the Heme Oxygenase with the bacteriophytochrome and the bottom band (blue box) is the plasmid backbone, providing evidence of successful composite construction.

http://25.media.tumblr.com/tumblr_mazgigGnrs1rg4kjpo1_500.jpg Pellets of cells containing induced heme oxygenase, uninducded heme oxygenase and lacking heme oxygenase

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1559
  Illegal BglII site found at 2550
  Illegal BamHI site found at 1919
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]