Part:BBa_K3168003
SmallBitNanoLuc
This basic part codes for the SmallBit of NanoLuc, which can be used as a split reporter. In combination with the LargeBit 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 SmallBit to enable the formation of fusion proteins. A strep-tag is also included at the end for protein purification. NanoLuc SmallBit variants with different affinities for LargeBit are available (Dixon, 2015). This basic part has a relatively low affinity (2.5*10-6 M) to prevent high background signals when combined with NanoLuc LargeBit. The affinity can be tuned by incorporating certain mutations (Dixon, 2015).
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 85
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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