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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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Plasmid
A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria and archaea; however plasmids are sometimes present in and eukaryotic organisms as well. Plasmids often carry useful genes, such as those involved in antibiotic resistance, virulence, secondary metabolism and bioremediation. While chromosomes are large and contain all the essential genetic information for living under normal conditions, plasmids are usually very small and contain additional genes for special circumstances. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. In the laboratory, plasmids may be introduced into a cell via transformation. Synthetic plasmids are available for procurement over the internet by various vendors ...
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Substrate Analog
Substrate analogs (substrate state analogues), are chemical compounds with a chemical structure that resemble the substrate molecule in an enzyme-catalyzed chemical reaction. Substrate analogs can act as competitive inhibitors of an enzymatic reaction. An example is phosphoramidate to the '' Tetrahymena'' group I ribozyme. Other examples of substrate analogs include 5’-adenylyl-imidodiphosphate, a substrate analog of ATP, and 3-acetylpyridine adenine dinucleotide, a substrate analog of NADH. As a competitive inhibitor, substrate analogs occupy the same binding site as its analog, and decrease the intended substrate’s efficiency. The maximum rate (Vmax) remains the same while the intended substrate’s affinity (measured by the Michaelis constant KM) is decreased. This means that less of the intended substrate will bind to the enzyme, resulting in less product being formed. In addition, the substrate analog may also be missing chemical components that allow the enzyme to g ...
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Escherichia Coli
''Escherichia coli'' ( )Wells, J. C. (2000) Longman Pronunciation Dictionary. Harlow ngland Pearson Education Ltd. is a gram-negative, facultative anaerobic, rod-shaped, coliform bacterium of the genus '' Escherichia'' that is commonly found in the lower intestine of warm-blooded organisms. Most ''E. coli'' strains are part of the normal microbiota of the gut, where they constitute about 0.1%, along with other facultative anaerobes. These bacteria are mostly harmless or even beneficial to humans. For example, some strains of ''E. coli'' benefit their hosts by producing vitamin K2 or by preventing the colonization of the intestine by harmful pathogenic bacteria. These mutually beneficial relationships between ''E. coli'' and humans are a type of mutualistic biological relationship—where both the humans and the ''E. coli'' are benefitting each other. ''E. coli'' is expelled into the environment within fecal matter. The bacterium grows massi ...
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Green Fluorescent Protein
The green fluorescent protein (GFP) is a protein that exhibits green fluorescence when exposed to light in the blue to ultraviolet range. The label ''GFP'' traditionally refers to the protein first isolated from the jellyfish ''Aequorea victoria'' and is sometimes called ''avGFP''. However, GFPs have been found in other organisms including corals, sea anemones, zoanithids, copepods and lancelets. The GFP from ''A. victoria'' has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. The fluorescence quantum yield (QY) of GFP is 0.79. The GFP from the sea pansy ('' Renilla reniformis'') has a single major excitation peak at 498 nm. GFP makes for an excellent tool in many forms of biology due to its ability to form an internal chromophore without requiring any accessory cofactors, gene products, or enzymes / substrates other than molecular ox ...
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GUS Reporter System
The GUS reporter system (''GUS'': β-glucuronidase) is a reporter gene system, particularly useful in plant molecular biology and microbiology. Several kinds of GUS reporter gene assay are available, depending on the substrate used. The term GUS staining refers to the most common of these, a Immunohistochemistry, histochemical technique. Purpose The purpose of this technique is to analyze the activity of a gene transcription Promoter (biology), promoter (in terms of gene expression, expression of a so-called reporter gene under the regulatory control of that promoter) either in a quantitative manner, involving some measure of activity, or qualitatively (on versus off) through visualization of its activity in different cells, Tissue (biology), tissues, or organs. The technique utilizes the uidA gene of ''Escherichia coli'', which codes for the enzyme, β-glucuronidase; this enzyme, when incubated with specific colorless or non-fluorescent Substrate (biology), substrates, can convert ...
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DsRed
Red fluorescent protein (RFP) is a protein which acts as a fluorophore, fluorescing red-orange when excited. The original variant occurs naturally in the coral genus ''Discosoma'', and is named DsRed. Several new variants have been developed using directed mutagenesis which fluoresce orange, red, and far-red. Characteristics and Properties Like GFP and other fluorescent proteins, RFP is a barrel-shaped protein made primarily out of Beta sheet, β-sheet motifs; this type of protein fold is commonly known as a Beta barrel, β-barrel. The mass of RFP is approximately 25.9 kDa. Its excitation maximum is 558 nm, and its emission maximum is 583 nm. Applications RFP is frequently used in molecular biology research as a fluorescent marker, for a variety of purposes. DsRed has been shown to be more suitable for optical imaging approaches than EGFP. Issues with fluorescent proteins include the length of time between protein synthesis and expression of fluorescence. DsRed ha ...
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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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Cell (biology)
The cell is the basic structural and functional unit of all life, forms of life. Every cell consists of cytoplasm enclosed within a Cell membrane, membrane; many cells contain organelles, each with a specific function. The term comes from the Latin word meaning 'small room'. Most cells are only visible under a light microscope, microscope. Cells Abiogenesis, emerged on Earth about 4 billion years ago. All cells are capable of Self-replication, replication, protein synthesis, and cell motility, motility. Cells are broadly categorized into two types: eukaryotic cells, which possess a Cell nucleus, nucleus, and prokaryotic, prokaryotic cells, which lack a nucleus but have a nucleoid region. Prokaryotes are single-celled organisms such as bacteria, whereas eukaryotes can be either single-celled, such as amoebae, or multicellular organism, multicellular, such as some algae, plants, animals, and fungi. Eukaryotic cells contain organelles including Mitochondrion, mitochondria, which ...
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Luminescence
Luminescence is a spontaneous emission of radiation from an electronically or vibrationally excited species not in thermal equilibrium with its environment. A luminescent object emits ''cold light'' in contrast to incandescence, where an object only emits light after heating. Generally, the emission of light is due to the movement of electrons between different energy levels within an atom after excitation by external factors. However, the exact mechanism of light emission in vibrationally excited species is unknown. The dials, hands, scales, and signs of aviation and navigational instruments and markings are often coated with luminescent materials in a process known as ''luminising''. Types * Ionoluminescence, a result of bombardment by fast ions * Radioluminescence, a result of bombardment by ionizing radiation * Electroluminescence, a result of an electric current passed through a substance ** Cathodoluminescence, a result of a luminescent material being struck by elect ...
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