Part:BBa_K1486021

Arabinose promoter + rLuc[1] + rLuc[2]

This part contains both fragments of the split renilla 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 Renilla 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

• rLuc[1] ( https://parts.igem.org/Part:BBa_K1486019 )
• rLuc[2] ( https://parts.igem.org/Part:BBa_K1486020 )
• CheY/CheZ + split rLuc ( https://parts.igem.org/Part:BBa_K1486054 )

References

S.K. Hatzios, S. Ringgaard, B. M. Davis, M. K. Waldor (2012, August 15). Studies of Dynamic Protein-Protein Interactions in Bacteria Using Renilla Luciferase Complementation Are Undermined by Nonspecific Enzyme Inhibition Plos One.

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 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