Part:BBa_K3168002

LargeBitNanoLuc

This basic part codes for the large bit of NanoLuc, which can be used as a split reporter. In combination with the small bit of NanoLuc and the addition of furimazine (the substrate), bright blue light is emitted (Figure 1). A flexible (GGS)₅ linker is located in front of the large bit to enable the formation of fusion proteins. A strep-tag is also included at the end for protein purification.

Figure 1. Split-NanoLuc principle.

Usage and Biology

Split reporters have been used a lot to detect protein-protein interactions and for screening purposes (Dixon, 2015). NanoLuc is a small, structurally robust and very bright luciferase. Many methods have been developed using the split variant of NanoLuc, such as the HiBiT technology and NanoLuc ternary technology (Ohmuro-Matsuyama, 2019).

References

Dixon, A. S., Schwinn, M. K., Hall, M. P., Zimmerman, K., Otto, P., Lubben, T. H., ... & Wood, M. G. (2015). NanoLuc complementation reporter optimized for accurate measurement of protein interactions in cells. ACS chemical biology, 11(2), 400-408.

Ohmuro-Matsuyama, Y., & Ueda, H. (2019). Protein-Protein Interaction Assays Using Split-NanoLuc. In Bioluminescence. IntechOpen.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 526
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]