Difference between revisions of "Part:BBa K1680009"
 
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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).
 
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).
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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. 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.  
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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.
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[[File:nanoluciferase.jpeg|300px|thumb|left|Figure 1 : Nanoluciferase reaction with its substrate called furimazine. The enzyme creates an oxidative process. This picture was taken on the Promega’s site.
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source : PDF https://promega.com>promega uk>teddy-riss-02
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]]
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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.
 
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.
 
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.
 
We codon-optimized the part for yeast. The NanoLuc is smaller than other luciferases while yielding equal or stronger luminescence readings.
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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 the reporter modules [https://parts.igem.org/Part:BBa_K5477030| BBa_K5477030] and [https://parts.igem.org/Part:BBa_K5477031| BBa_K5477031] to build the following devices [https://parts.igem.org/Part:BBa_K5477041| BBa_K5477041], [https://parts.igem.org/Part:BBa_K5477042| BBa_K5477042], [https://parts.igem.org/Part:BBa_K5477043| BBa_K5477043], [https://parts.igem.org/Part:BBa_K5477044| BBa_K5477044], [https://parts.igem.org/Part:BBa_K5477045| BBa_K5477045] and [https://parts.igem.org/Part:BBa_K5477046| BBa_K5477046].
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 21:16, 1 October 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 the reporter modules BBa_K5477030 and BBa_K5477031 to build the following devices BBa_K5477041, BBa_K5477042, BBa_K5477043, BBa_K5477044, BBa_K5477045 and BBa_K5477046.


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]