Part:BBa_K1680009

NanoLuc Luciferase

This part contains the protein coding region for the NanoLuc(tm) luciferase, which is originally designed by Promega Corporation (http://bit.ly/1Vgmab3, free for research use). Nanoluciferase is a small and monomeric enzyme (19.1kDa) and is about 150 times brighter than other luciferases. The new substrate called furimazine produces a high intensity luminescence.

Figure 1 : Nanoluciferase reaction with its substrate called furimazine. The enzyme creates an oxidative process. This picture was taken on the Promega’s site. source : PDF https://promega.com>promega uk>teddy-riss-02

After looking at the bibliography, Nanoluciferase gene was chosen because it allowed us to have a stronger signal. Indeed, this protein isn’t secreted but unfused. Furthermore, its intracellular half-life is superior to 6 hours and the Nanoluciferase has a low autoluminescence. This gives us maximum sensitivity. NLuc is used in biomedical research for several applications, including studying protein-protein interactions, monitoring protein stability, BRET-based sensors, molecular imaging or investigating genetic regulation and cell signaling. The emission pic of nanoluciferase is located at 460nm. We codon-optimized the part for yeast. The NanoLuc is smaller than other luciferases while yielding equal or stronger luminescence readings.

RLU measurement of the NanoLuc under pSUC2 promoter part:BBa_K950003 control, normalized to OD600.

RLU measurement of the NanoLuc under pFET3 promoter part:BBa_K950000 control, normalized to OD600.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]