Renilla luciferase, a monomeric 36kDa protein, catalyzes coelenterate-luciferin (coelenterazine) oxidation to produce light (3; Figure 1). Post-translational modification is not required for its activity, and the enzyme may function as a genetic reporter immediately following translation.
What is the purpose of luciferase?
The power of luciferase has been harnessed by scientists to devise reactions whose light output is used to monitor biological processes including gene expression, biomolecular binding, and cell viability.17 Nov 2020
How can you use a luciferase assay to determine gene activity?
A luciferase assay. To determine if a protein is able to activate (or suppress) transcription of a gene of interest, a scientist uses recombinant DNA technology to produce a construct in which the gene's promoter is placed adjacent to a luciferase reporter gene.
What is TK Renilla vector?
The pRL Vectors are wild-type Renilla luciferase (Rluc) control reporter vectors. The HSV-thymidine kinase promoter (pRL-TK) is relatively weak and may be particularly useful in providing neutral constitutive expression of the Renilla luciferase control reporter.
How does luciferase activity work?
The luciferase assay is useful to study whether a protein of interest regulates a particular gene at the transcription level. When this protein activates transcription, the cell will produce luciferase enzyme. After the addition of a lysis buffer and a substrate, a luminometer quantifies the luciferase activity.
How does the gene luciferase work how does it produce light?
Fireflies produce a chemical reaction inside their bodies that allows them to light up. When oxygen combines with calcium, adenosine triphosphate (ATP) and the chemical luciferin in the presence of luciferase, a bioluminescent enzyme, light is produced.5 Sept 2005
What is a luciferase gene?
Luciferases are enzymes that produce light when they oxidize their substrate. The gene for the most common luciferase comes from the firefly, but luciferases from other animals such as the sea pansy Renilla reniformis, the copepod Gaussia princeps, and the ostracod Cypridina noctiluca are also used as reporters.
What is the purpose of a dual luciferase assay?
The Dual Luciferase (Firefly-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly.The Dual Luciferase (FireflyLuciferase (FireflyThe chemical reaction catalyzed by firefly luciferase takes place in two steps: luciferin + ATP → luciferyl adenylate + PP. luciferyl adenylate + O2 → oxyluciferin + AMP + light.https://en.wikipedia.org › wiki › Firefly_luciferaseFirefly luciferase - Wikipedia-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly.
What is firefly luciferase assay?
The Firefly Luciferase HTS Assay (SCT150) is designed with a proprietary mixture of substances that modify the enzymatic reaction to produce a long-lasting signal (steady-glow) by preventing the formation of adenyl-oxyluciferin at the enzyme surface.
How does the gene luciferase work?
A commonly used reporter gene is the luciferase gene from the firefly Photinus pyralis. This gene encodes a 61-kDa enzyme that oxidizes D-luciferin in the presence of ATP, oxygen, and Mg(++), yielding a fluorescent product that can be quantified by measuring the released light.
What kind of protein is luciferase?
Fluorescent Proteins 101: Luciferase. Luciferases are a class of enzymes capable of catalyzing chemical reactions in living organisms resulting in the emission of photons.24 Jun 2014
What is a luciferase enzyme?
Luciferases are enzymes that use a substrate called luciferin, along with oxygen and ATP, in an energetic process that produces light—like the yellow glow of fireflies.17 Nov 2020
Is luciferase a protein?
Luciferases are proteins with enzymatic activity that, in the presence of ATP, oxygen, and the appropriate substrate (typically luciferin), catalyze the oxidation of the substrate in a reaction that results in the emission of a photon.