Difference between revisions of "Part:BBa K1616023"
| (2 intermediate revisions by the same user not shown) | |||
| Line 2: | Line 2: | ||
<partinfo>BBa_K1616023 short</partinfo> | <partinfo>BBa_K1616023 short</partinfo> | ||
<br><br> | <br><br> | ||
| + | <h3> <font style="color:#b22222">NanoLuc</font> </h3> | ||
NanoLuc (Nluc) is a novel engineered luciferase coupled with a novel coelenterazine analogue: Furimazine. When both are coupled, we obtain a much brighter luminescence than Fluc or Rluc and it is known to work exceptionally well as a reporter. | NanoLuc (Nluc) is a novel engineered luciferase coupled with a novel coelenterazine analogue: Furimazine. When both are coupled, we obtain a much brighter luminescence than Fluc or Rluc and it is known to work exceptionally well as a reporter. | ||
NanoLuc is a 19.1 kDa enzyme which is an ATP independent luciferase providing a glow-type luminescence. | NanoLuc is a 19.1 kDa enzyme which is an ATP independent luciferase providing a glow-type luminescence. | ||
| − | Thermal stable enzyme | + | <b>Thermal stable enzyme</b><br> |
| − | NanoLuc retains activity following 30 min incubation at 55 °C. | + | NanoLuc retains activity following 30 min incubation at 55 °C. <br> |
| − | Active over broad pH range | + | <b>Active over broad pH range</b><br> |
| − | NanoLuc is fully active between pH 7-9 and retains significant activity at pH 5-7 | + | NanoLuc is fully active between pH 7-9 and retains significant activity at pH 5-7 <br> |
| − | Monomeric enzyme | + | <b>Monomeric enzyme</b><br> |
NanoLuc facilitates use as transcriptional reporter or fusion partner. | NanoLuc facilitates use as transcriptional reporter or fusion partner. | ||
| − | + | <br> | |
<center> | <center> | ||
[[Image:Nanoluc reaction copie.png|600px|]] | [[Image:Nanoluc reaction copie.png|600px|]] | ||
</center> | </center> | ||
| + | <h3> <font style="color:#b22222">A fast detection system involving chemical processes </font> </h3> | ||
| + | One other method of fast detection is bioluminescence. This trait found in several marine bacteria emits light by the mean of biochemical mechanisms involving chemical processes. It is important to make the distinction between fluorescence and bioluminescence.In fluorescence, a high energy photon excites a molecule to an excited state which makes the latter lose energy. This loss leads to the release of a visible photon and the drop to the original groundstate of the molecule. Bioluminescence involves an oxygen oxidation of an organic molecule. A luciferase catalyzes the oxidation of luciferin to excited state oxyluciferin which release a visible photon (Shreeram Akilesh, Bioluminescence: Nature’s Bright Idea,2000). | ||
| + | <br><br> | ||
| + | <h3> <font style="color:#b22222"> Luciferase: an essential enzyme for bioluminescence </font> </h3> | ||
| + | Firefly (Fluc, 61 kDa) and Renilla (Rluc, 36 kDa) luciferases have been used for a huge amount of applications such as for the study of molecular and cellular processes. Given this fact, researchers tried to investigate alternative luciferases from marine organims and achieve a more optimal enzyme structure which could explore other variants of luminogenic substrate. Furthermore, as most of the luciferases come from marine organisms, the substrate the most used is coelenterazine. | ||
| − | + | ||
| − | <br> | + | <br><br> |
===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K1616023 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1616023 SequenceAndFeatures</partinfo> | ||
| Line 23: | Line 29: | ||
===Design Notes=== | ===Design Notes=== | ||
| − | + | All BioBrick parts used for assembling the composite part are compatible with the RFC10 Biobrick standard. | |
===Source=== | ===Source=== | ||
Latest revision as of 01:27, 23 September 2015
T7 promoter - HisTag- NanoLuc - T7 terminator
NanoLuc
NanoLuc (Nluc) is a novel engineered luciferase coupled with a novel coelenterazine analogue: Furimazine. When both are coupled, we obtain a much brighter luminescence than Fluc or Rluc and it is known to work exceptionally well as a reporter. NanoLuc is a 19.1 kDa enzyme which is an ATP independent luciferase providing a glow-type luminescence.
Thermal stable enzyme
NanoLuc retains activity following 30 min incubation at 55 °C.
Active over broad pH range
NanoLuc is fully active between pH 7-9 and retains significant activity at pH 5-7
Monomeric enzyme
NanoLuc facilitates use as transcriptional reporter or fusion partner.
A fast detection system involving chemical processes
One other method of fast detection is bioluminescence. This trait found in several marine bacteria emits light by the mean of biochemical mechanisms involving chemical processes. It is important to make the distinction between fluorescence and bioluminescence.In fluorescence, a high energy photon excites a molecule to an excited state which makes the latter lose energy. This loss leads to the release of a visible photon and the drop to the original groundstate of the molecule. Bioluminescence involves an oxygen oxidation of an organic molecule. A luciferase catalyzes the oxidation of luciferin to excited state oxyluciferin which release a visible photon (Shreeram Akilesh, Bioluminescence: Nature’s Bright Idea,2000).
Luciferase: an essential enzyme for bioluminescence
Firefly (Fluc, 61 kDa) and Renilla (Rluc, 36 kDa) luciferases have been used for a huge amount of applications such as for the study of molecular and cellular processes. Given this fact, researchers tried to investigate alternative luciferases from marine organims and achieve a more optimal enzyme structure which could explore other variants of luminogenic substrate. Furthermore, as most of the luciferases come from marine organisms, the substrate the most used is coelenterazine.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 117
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 20
Design Notes
All BioBrick parts used for assembling the composite part are compatible with the RFC10 Biobrick standard.
Source
NanoLuc is a 19.1 kDa enzyme which is an ATP independent luciferase providing a glow-type luminescence.
References
Kyle Hooper, Ph.D, Applications of a smaller, brighter, more versatile luciferase:NanoLuc™ Luciferase Technology, 2012
Shreeram Akilesh, Bioluminescence: Nature’s Bright Idea,2000.