Part:BBa_K953006

Heme Oxygenase with T7 + Deinococcus radiodurans Bacteriophytochrome (E. coli Codon Optimised)

This composite couples Synechocystis Heme Oxygenase with T7 (Part BBa_K953000) and Deinococcus radiodurans bacteriophytochrome (Part BBa_K953001), 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.

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) 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 314
  Illegal BglII site found at 1713
  Illegal BamHI site found at 1518
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1062
  Illegal NgoMIV site found at 1131
  Illegal NgoMIV site found at 2052
  Illegal AgeI site found at 1318
  Illegal AgeI site found at 1363
  Illegal AgeI site found at 2424
  
• 1000
  COMPATIBLE WITH RFC[1000]