Difference between revisions of "Part:BBa K1680009"

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.

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 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]

@@ Line 2: / Line 2: @@
 <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). We codon-optimized the part for yeast. The NanoLuc is smaller than other luciferases while yielding equal or stronger luminescence readings.
+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.
+[[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
+]]
+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.
@@ Line 15: / Line 38: @@
+<h2> MilkClear iGEM UCopenhagen 2024 </h2>
+Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/
+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].
 <!-- Add more about the biology of this part here