Coding

Part:BBa_K1680009

Designed by: Katharina Sporbeck   Group: iGEM15_Tuebingen   (2015-09-17)
Revision as of 13:24, 8 October 2019 by Rollom (Talk | contribs)

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.






PSUC-conc.png

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

Team Tuebingen mpr 6h.jpg

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]


[edit]
Categories
//cds
//cds/reporter
//chassis/eukaryote/yeast
//function/reporter/light
Parameters
None