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Luciferase
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. Both words are derived from the Latin word ''lucifer'', meaning "lightbearer", which in turn is derived from the Latin words for "light" (''lux)'' and "to bring or carry" (''ferre)''.Luciferases are widely used in biotechnology, for bioluminescence imaging microscopy and as reporter genes, for many of the same applications as fluorescent proteins. However, unlike fluorescent proteins, luciferases do not require an external light source, but do require addition of luciferin, the consumable substrate. Examples A variety of organisms regulate their light production using different luciferases in a variety of light-emitting reactions. The majority of studied luciferases have been found in anima ...
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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. Mechanism of reaction The chemical reaction catalyzed by firefly luciferase takes place in two steps: * luciferin + ATP → luciferyl adenylate + PPi * luciferyl adenylate + O2 → oxyluciferin + AMP + light Light is produced because the reaction forms oxyluciferin in an electronically excited state. The reaction releases a photon of light as oxyluciferin goes back to the ground state. Luciferyl adenylate can additionally participate in a side reaction with O2 to form hydrogen peroxide and dehydroluciferyl-AMP. About 20% of the luciferyl adenylate intermediate is oxidized in this pathway. Firefly luciferase generates light from luciferin in a multistep process. First, D-lu ...
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Bioluminescent Bacteria
Bioluminescent bacteria are Bioluminescence, light-producing bacteria that are predominantly present in sea water, marine sediments, the surface of decomposing fish and in the gut of marine animals. While not as common, bacterial bioluminescence is also found in terrestrial and freshwater bacteria. These bacteria may be free living (such as ''Vibrio harveyi'') or in symbiosis with animals such as the Hawaiian Bobtail Squid, Hawaiian Bobtail squid (''Aliivibrio fischeri'') or terrestrial nematodes (''Photorhabdus luminescens''). The host organisms provide these bacteria a safe home and sufficient nutrition. In exchange, the hosts use the light produced by the bacteria for camouflage, prey and/or mate attraction. Bioluminescent bacteria have evolved symbiotic relationships with other organisms in which both participants benefit each other equally. Bacteria also use luminescence reaction for quorum sensing, an ability to regulate gene expression in response to bacterial cell density. ...
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Bioluminescence
Bioluminescence is the emission of light during a chemiluminescence reaction by living organisms. Bioluminescence occurs in multifarious organisms ranging from marine vertebrates and invertebrates, as well as in some Fungus, fungi, microorganisms including some bioluminescent bacteria, Dinoflagellate, dinoflagellates and terrestrial arthropods such as Firefly, fireflies. In some animals, the light is bacteriogenic, produced by symbiosis, symbiotic bacteria such as those from the genus ''Vibrio''; in others, it is autogenic, produced by the animals themselves. In most cases, the principal chemical reaction in bioluminescence involves the reaction of a substrate called luciferin and an enzyme, called luciferase. Because these are generic names, luciferins and luciferases are often distinguished by the species or group, e.g. firefly luciferin or Vargulin, cypridina luciferin. In all characterized cases, the enzyme Catalysis, catalyzes the Redox, oxidation of the luciferin resultin ...
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Bioluminescence Imaging
Bioluminescence imaging (BLI) is a technology developed over the past decades (1990's and onward). that allows for the noninvasive study of ongoing biological processes Recently, bioluminescence tomography (BLT) has become possible and several systems have become commercially available. In 2011, PerkinElmer acquired one of the most popular lines of optical imaging systems with bioluminescence from Caliper Life Sciences. Background Bioluminescence is the process of light emission in living organisms. Bioluminescence imaging utilizes native light emission from one of several organisms which bioluminesce, also known as luciferase enzymes. The three main sources are the North American firefly, the sea pansy (and related marine organisms), and bacteria like '' Photorhabdus luminescens'' and '' Vibrio fischeri''. The DNA encoding the luminescent protein is incorporated into the laboratory animal either via a viral vector or by creating a transgenic animal. Rodent models of cancer ...
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Renilla-luciferin 2-monooxygenase
Renilla-luciferin 2-monooxygenase, ''Renilla'' luciferase, or RLuc, is a bioluminescent enzyme found in '' Renilla reniformis'', belonging to a group of coelenterazine luciferases. Of this group of enzymes, the luciferase from '' Renilla reniformis'' has been the most extensively studied, and due to its bioluminescence requiring only molecular oxygen, has a wide range of applications, with uses as a reporter gene probe in cell culture, in vivo imaging, and various other areas of biological research. Recently, chimeras of RLuc have been developed and demonstrated to be the brightest luminescent proteins to date, and have proved effective in both noninvasive single-cell and whole body imaging. Note that the EC record also includes other unrelated enzymes that catalyze the same reaction. An example is the calcium-dependent photoprotein aequorin: while Rluc is in the AB hydrolase superfamily, aequorin is an EF hand protein. The name does not specifically refer to ''Renilla'', but in ...
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Luciferin
Luciferin () is a generic term for the light-emitting chemical compound, compound found in organisms that generate bioluminescence. Luciferins typically undergo an enzyme-catalyzed reaction with Oxygen, molecular oxygen. The resulting transformation, which usually involves breaking off a molecular fragment, produces an excited state intermediate that emits light upon decaying to its ground state. The term may refer to molecules that are substrates for both luciferases and photoproteins. Types Luciferins are a class of small-molecule substrate (biochemistry), substrates that react with oxygen in the presence of a luciferase (an enzyme) to release energy in the form of light. It is not known just how many types of luciferins there are, but some of the better-studied compounds are listed below. Because of the chemical diversity of luciferins, there is no clear unifying mechanism of action, except that all require molecular oxygen, The variety of luciferins and luciferases, their ...
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Reporter Gene
Reporter genes are molecular tools widely used in molecular biology, genetics, and biotechnology to study gene function, expression patterns, and regulatory mechanisms. These genes encode proteins that produce easily detectable signals, such as fluorescence, luminescence, or enzymatic activity, allowing researchers to monitor cellular processes in real-time. Reporter genes are often fused to regulatory sequences of genes of interest, enabling scientists to analyze promoter activity, transcriptional regulation, and signal transduction pathways. Common reporter gene systems include green fluorescent protein (GFP), β-galactosidase (lacZ), luciferase, and chloramphenicol acetyltransferase (CAT), each offering distinct advantages depending on the experimental application. Their versatility makes reporter genes invaluable in fields such as drug discovery, gene therapy, and synthetic biology. Common Reporter Genes To introduce a reporter gene into an organism, scientists place the ...
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Coelenterazine
Coelenterazine is a luciferin, a molecule that emits light after reaction with oxygen, found in many aquatic organisms across eight phyla. It is the substrate of many luciferases such as '' Renilla reniformis'' luciferase (Rluc), ''Gaussia'' luciferase (Gluc), and photoproteins, including aequorin, and obelin. All these proteins catalyze the oxidation of this substance, a reaction catalogued EC 1.13.12.5. History Coelenterazine was simultaneously isolated and characterized by two groups studying the luminescent organisms sea pansy ('' Renilla reniformis'') and the cnidarian ''Aequorea victoria'', respectively. Both groups independently discovered that the same compound was used in both luminescent systems. The molecule was named after the now-obsolete phylum coelenterata. Likewise, the two main metabolites – coelenteramide and coelenteramine – were named after their respective functional groups. While coelenterazine was first discovered in ''Aequorea victoria'', it wa ...
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Photinus Pyralis
''Photinus pyralis'', also known by the common names the common eastern firefly or big dipper firefly, and sometimes called a "lightning bug", is a species of flying beetle. An organ on its abdomen is responsible for its light production. It is the most common species of firefly in North America, and is typically found east of the Rocky Mountains. ''Photinus'' fireflies are often confused with fireflies of the similar-sounding genus, ''Photuris,'' which are also found in North America. These fireflies are most noticeable around twilight in the spring and summer months.Mast, S.O. 1912. Behaviour of fire-flies (Photinus pyralis) with special references to the problem of orientation. 256-272 The common name, big dipper firefly, is due to the characteristic flight of the males, whose trajectory appears to follow a J-shape, lighting on the upswing. During flight, this J-pattern is used alongside light flashing to attract females, who rest on vegetation and signal back to males if inte ...
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Photoprotein
Photoproteins are a type of enzyme produced by bioluminescent organisms. They add to the function of the luciferins whose usual light-producing reaction is catalyzed by the enzyme luciferase. History The term photoprotein was first used to describe the unusual chemistry of the luminescent system of ''Chaetopterus'' (a marine Polychaete worm). This was meant to distinguish them from other light-producing proteins because these do not exhibit the usual luciferin-luciferase reaction. Reaction kinetics Photoproteins do not display typical enzyme kinetics as seen in luciferases. Instead, when mixed with luciferin, they display luminescence proportional to the amount of the photoprotein. For example, the photoprotein aequorin produces a flash of light when luciferin and calcium are added, rather than the prolonged glow that is seen for luciferases when luciferin is added. In this respect, it may appear that photoproteins are not enzymes, when in fact they do catalyze their biolumines ...
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Photinini
The Photinini are a large tribe of fireflies in the subfamily Lampyrinae. '' Photinus pyralis'' is famous in biotechnology for its luciferase gene. This is sometimes employed as a marker gene; genetically modified organisms which contain it start to glow like the firefly when brought in contact with a luciferin-containing medium. Firefly luciferases differ slightly between taxa, resulting in differently colored light and other properties, and in most cases where "firefly luciferase" is used in some application or study, it is the specific luciferase of '' P. pyralis''. Systematics The group has recently been examined using molecular phylogenetics, using fairly comprehensive sampling. Genera * '' Ankonophallus'' Zaragoza-Caballero & Navarrete-Heredia, 2014 * '' Aorphallus'' Zaragoza-Caballero & Gutierrez-Carranza, 2018 * '' Callopisma'' Motschulsky, 1853 * '' Calotrechelum'' Pic, 1930 * '' Dadophora'' Olivier, 1907 * '' Dilychnia'' Motschulsky, 1853 * ''Ellychnia'' LeConte, 18 ...
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Dinoflagellates
The Dinoflagellates (), also called Dinophytes, are a monophyletic group of single-celled eukaryotes constituting the phylum Dinoflagellata and are usually considered protists. Dinoflagellates are mostly marine plankton, but they are also common in freshwater habitats. Their populations vary with sea surface temperature, salinity, and depth. Many dinoflagellates are photosynthetic, but a large fraction of these are in fact mixotrophic, combining photosynthesis with ingestion of prey ( phagotrophy and myzocytosis). In terms of number of species, dinoflagellates are one of the largest groups of marine eukaryotes, although substantially smaller than diatoms. Some species are endosymbionts of marine animals and play an important part in the biology of coral reefs. Other dinoflagellates are unpigmented predators on other protozoa, and a few forms are parasitic (for example, '' Oodinium'' and '' Pfiesteria''). Some dinoflagellates produce resting stages, called dinoflagellate cysts ...
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