What gene makes the luciferase enzyme?
What gene makes the luciferase enzyme?
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 is the firefly gene?
Abstract. The firefly luciferase gene (luc) from Photinus pyralis is a new tool for visualizing gene expression in plants. Luciferase (LUC) catalyzes the ATP-dependent oxidative decarboxylation of luciferin, a reaction that produces light.
How the expression of insert DNA can be measured by luciferase activity?
To measure luciferase activity of the cell lysates, you will need a multiwell plate or a tube containing cell lysates and a luminometer. This apparatus detects the photon emission produced from the luciferase reaction and the unit of measurement comes out as relative light units (RLU).
Who made luciferase?
Raphaël Dubois
Luciferase is a generic term for the class of oxidative enzymes that produce bioluminescence, and is usually distinguished from a photoprotein. The name was first used by Raphaël Dubois who invented the words luciferin and luciferase, for the substrate and enzyme, respectively.
How is luciferase produced in a firefly?
When oxygen combines with calcium, adenosine triphosphate (ATP) and the chemical luciferin in the presence of luciferase, a bioluminescent enzyme, light is produced. Unlike a light bulb, which produces a lot of heat in addition to light, a firefly’s light is “cold light” without a lot of energy being lost as heat.
What is the function of firefly luciferase?
Firefly luciferase is the light-emitting enzyme responsible for the bioluminescence of fireflies and click beetles. The enzyme catalyses the oxidation of firefly luciferin, requiring oxygen and ATP. Because of the requirement of ATP, firefly luciferases have been used extensively in biotechnology.
Is luciferase monomeric or dimeric?
Firefly luciferase is extremely hydrophobic; the monomer is apparently active, but the enzyme exists as a dimer in most solutions [6].
How does dual luciferase assay work?
The dual luciferase assay is performed by sequentially measuring the firefly and Renilla luciferase activities of the same sample, with the results expressed as the ratio of firefly to Renilla luciferase activity (Fluc/Rluc).
