Difference between revisions of "Part:BBa K1680009"

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<h2> MilkClear iGEM UCopenhagen 2024 </h2>
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Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/
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This part was used in our biosensor, specifically in our reporter module [https://parts.igem.org/Part:BBa_K5477030| BBa_K5477030] and [https://parts.igem.org/Part:BBa_K5477031| BBa_K5477031].
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 20:26, 26 September 2024

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.


MilkClear iGEM UCopenhagen 2024

Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/

This part was used in our biosensor, specifically in our reporter module BBa_K5477030 and BBa_K5477031.


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]