Difference between revisions of "Part:BBa K1680009"
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<partinfo>BBa_K1680009 short</partinfo> | <partinfo>BBa_K1680009 short</partinfo> | ||
− | + | 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. | ||
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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. | ||
+ | source : PDF https://promega.com>promega uk>teddy-riss-02 | ||
+ | ]] | ||
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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. | ||
+ | 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. | ||
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+ | [[Image:PSUC-conc.png|400px]] | ||
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+ | RLU measurement of the NanoLuc under pSUC2 promoter [[part:BBa_K950003]] control, normalized to OD600. | ||
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+ | [[Image:Team Tuebingen mpr 6h.jpg|400px]] | ||
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+ | RLU measurement of the NanoLuc under pFET3 promoter [[part:BBa_K950000]] control, normalized to OD600. | ||
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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 | ||
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− | <span class='h3bb'>Sequence and Features</span> | + | <span class='h3bb'>'''Sequence and Features'''</span> |
<partinfo>BBa_K1680009 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1680009 SequenceAndFeatures</partinfo> | ||
Latest 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.
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.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]