Part:BBa_K1486018

Arabinose promoter + fLuc[1] + fLuc[2]

This part contains both fragments of the split firefly luciferase, each of them preceded by an Elowitz RBS.

Purpose of the Biobrick

This construct aimed to establish a background noise for luciferase complementation assays, in order to substract it to the complementation constructs' signal.

Experiment: CheY CheZ complementation assay

An experiment of bioluminescence assay to study the interaction between CheY and CheZ chemotaxis associated proteins in E.Coli has been done, with this split Firefly luciferase as a negative control. The experiment didn't work well for the CheY CheZ construct, but we could establish that the signal of the full luciferase was 100 fold higher than the split.

Associated Biobricks

• fLuc[1] ( https://parts.igem.org/Part:BBa_K1486016 )
• fLuc[2] ( https://parts.igem.org/Part:BBa_K1486017 )
• CheY/CheZ + split fLuc ( https://parts.igem.org/Part:BBa_K1486055 )

References

Luker KE1, Smith MC, Luker GD, Gammon ST, Piwnica-Worms H, Piwnica-Worms D., (August 17, 2004), Kinetics of regulated protein-protein interactions revealed with firefly luciferase complementation imaging in cells and living animals

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1205
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1653
  Illegal BamHI site found at 1144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961
  Illegal SapI.rc site found at 1357