GAL4 UAS System
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GAL4 UAS System
The Gal4 transcription factor is a positive regulator of gene expression of galactose-induced genes. This protein represents a large fungal family of transcription factors, Gal4 family, which includes over 50 members in the yeast ''Saccharomyces cerevisiae'' e.g. Oaf1, Pip2, Pdr1, Pdr3, Leu3. Gal4 recognizes genes with UAS, an upstream activating sequence, and activates them. In yeast cells, the principal targets are GAL1 (galactokinase), GAL10 ( UDP-glucose 4-epimerase), and GAL7 (galactose-1-phosphate uridylyltransferase), three enzymes required for galactose metabolism. This binding has also proven useful in constructing the GAL4/UAS system, a technique for controlling expression in insects. In yeast, Gal4 is by default repressed by Gal80, and activated in the presence of galactose as Gal3 binds away Gal80. Domains Two executive domains, DNA binding and activation domains, provide key function of the Gal4 protein conforming to most of the transcription factors. DNA bindi ...
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Saccharomyces Cerevisiae
''Saccharomyces cerevisiae'' () (brewer's yeast or baker's yeast) is a species of yeast (single-celled fungus microorganisms). The species has been instrumental in winemaking, baking, and brewing since ancient times. It is believed to have been originally isolated from the skin of grapes. It is one of the most intensively studied eukaryotic model organisms in molecular biology, molecular and cell biology, much like ''Escherichia coli'' as the model bacteria, bacterium. It is the microorganism behind the most common type of fermentation (biochemistry), fermentation. ''S. cerevisiae'' cells are round to ovoid, 5–10 micrometre, μm in diameter. It reproduces by budding. Many proteins important in human biology were first discovered by studying their Homology (biology), homologs in yeast; these proteins include cell cycle proteins, signaling proteins, and protein-processing enzymes. ''S. cerevisiae'' is currently the only yeast cell known to have Berkeley body, Berkeley bo ...
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Two-hybrid Screening
Two-hybrid screening (originally known as yeast two-hybrid system or Y2H) is a molecular biology technique used to discover protein–protein interactions (PPIs) and protein–DNA interactions by testing for physical interactions (such as binding) between two proteins or a single protein and a DNA molecule, respectively. The premise behind the test is the activation of downstream reporter gene(s) by the binding of a transcription factor onto an upstream activating sequence (UAS). For two-hybrid screening, the transcription factor is split into two separate fragments, called the DNA-binding domain (DBD or often also abbreviated as BD) and activating domain (AD). The BD is the domain responsible for binding to the UAS and the AD is the domain responsible for the activation of transcription. The Y2H is thus a protein-fragment complementation assay. History Pioneered by Stanley Fields and Ok-Kyu Song in 1989, the technique was originally designed to detect protein–protein ...
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Leavening Agents
In cooking, a leavening agent () or raising agent, also called a leaven () or leavener, is any one of a number of substances used in doughs and batters that cause a foaming action (gas bubbles) that lightens and softens the mixture. An alternative or supplement to leavening agents is mechanical action by which air is incorporated (i.e. kneading). Leavening agents can be biological or synthetic chemical compounds. The gas produced is often carbon dioxide, or occasionally hydrogen. When a dough or batter is mixed, the starch in the flour and the water in the dough form a matrix (often supported further by proteins like gluten or polysaccharides, such as pentosans or xanthan gum). The starch then gelatinizes and sets, leaving gas bubbles that remain. Biological leavening agents * ''Saccharomyces cerevisiae'' producing carbon dioxide found in: ** baker's yeast ** Beer barm (unpasteurised—live yeast) ** ginger beer ** kefir ** sourdough starter * ''Clostridium perfringens'' ...
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Yeasts Used In Brewing
Yeasts are eukaryotic, single-celled microorganisms classified as members of the fungus kingdom. The first yeast originated hundreds of millions of years ago, and at least 1,500 species are currently recognized. They are estimated to constitute 1% of all described fungal species. Yeasts are unicellular organisms that evolved from multicellular ancestors, with some species having the ability to develop multicellular characteristics by forming strings of connected budding cells known as pseudohyphae or false hyphae. Yeast sizes vary greatly, depending on species and environment, typically measuring 3–4  µm in diameter, although some yeasts can grow to 40 µm in size. Most yeasts reproduce asexually by mitosis, and many do so by the asymmetric division process known as budding. With their single-celled growth habit, yeasts can be contrasted with molds, which grow hyphae. Fungal species that can take both forms (depending on temperature or other conditions) are called ...
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Osmophiles
An osmophile is a microorganism adapted to environments with high osmotic pressures, such as high sugar concentrations. Osmophiles are similar to halophiles (salt-loving organisms) in that a critical aspect of both types of environment is their low water activity, aW. High sugar concentrations represent a growth-limiting factor for many microorganisms, yet osmophiles protect themselves against this high osmotic pressure by the synthesis of osmoprotectants such as alcohols and amino acids. Many osmophilic microorganisms are yeast Yeasts are eukaryotic, single-celled microorganisms classified as members of the fungus kingdom. The first yeast originated hundreds of millions of years ago, and at least 1,500 species are currently recognized. They are estimated to constitut ...s; some bacteria are also osmophilic. Osmophilic yeasts are important because they cause spoilage in the sugar and sweet goods industry, with products such as fruit juices, fruit juice concentrates, liquid sug ...
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Probiotics
Probiotics are live microorganisms promoted with claims that they provide health benefits when consumed, generally by improving or restoring the gut microbiota. Probiotics are considered generally safe to consume, but may cause bacteria-host interactions and unwanted side effects in rare cases. There is some evidence that probiotics are beneficial for some conditions, but there is little evidence for many of the health benefits claimed for them. The first discovered probiotic was a certain strain of bacillus in Bulgarian yoghurt, called ''Lactobacillus bulgaricus''. The discovery was made in 1905 by Bulgarian physician and microbiologist Stamen Grigorov. The modern-day theory is generally attributed to Russian Nobel laureate Élie Metchnikoff, who postulated around 1907 that yoghurt-consuming Bulgarian peasants lived longer. A growing probiotics market has led to the need for stricter requirements for scientific substantiation of putative benefits conferred by microorganisms ...
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Digestive System
The human digestive system consists of the gastrointestinal tract plus the accessory organs of digestion (the tongue, salivary glands, pancreas, liver, and gallbladder). Digestion involves the breakdown of food into smaller and smaller components, until they can be absorbed and assimilated into the body. The process of digestion has three stages: the cephalic phase, the gastric phase, and the intestinal phase. The first stage, the cephalic phase of digestion, begins with secretions from gastric glands in response to the sight and smell of food. This stage includes the mechanical breakdown of food by chewing, and the chemical breakdown by digestive enzymes, that takes place in the mouth. Saliva contains the digestive enzymes amylase, and lingual lipase, secreted by the salivary and serous glands on the tongue. Chewing, in which the food is mixed with saliva, begins the mechanical process of digestion. This produces a bolus which is swallowed down the esophagus to enter the st ...
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Fungal Models
A fungus ( : fungi or funguses) is any member of the group of eukaryotic organisms that includes microorganisms such as yeasts and molds, as well as the more familiar mushrooms. These organisms are classified as a kingdom, separately from the other eukaryotic kingdoms, which by one traditional classification include Plantae, Animalia, Protozoa, and Chromista. A characteristic that places fungi in a different kingdom from plants, bacteria, and some protists is chitin in their cell walls. Fungi, like animals, are heterotrophs; they acquire their food by absorbing dissolved molecules, typically by secreting digestive enzymes into their environment. Fungi do not photosynthesize. Growth is their means of mobility, except for spores (a few of which are flagellated), which may travel through the air or water. Fungi are the principal decomposers in ecological systems. These and other differences place fungi in a single group of related organisms, named the ''Eumycota'' (''true f ...
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Sugar Substitutes
A sugar substitute is a food additive that provides a sweetness like that of sugar while containing significantly less food energy than sugar-based sweeteners, making it a zero-calorie () or low-calorie sweetener. Artificial sweeteners may be derived through manufacturing of plant extracts or processed by chemical synthesis. Sugar substitute products are commercially available in various forms, such as small pills, powders, and packets. In North America, common sugar substitutes include aspartame, monk fruit extract, saccharin, sucralose, and stevia; cyclamate is also used outside the United States. These sweeteners are a fundamental ingredient in diet drinks to sweeten them without adding calories. Additionally, sugar alcohols such as erythritol, xylitol, and sorbitol are derived from sugars. Approved artificial sweeteners do not cause cancer. Reviews and dietetic professionals have concluded that moderate use of non-nutritive sweeteners as a safe replacement for sugars can ...
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Nutrition
Nutrition is the biochemical and physiological process by which an organism uses food to support its life. It provides organisms with nutrients, which can be metabolized to create energy and chemical structures. Failure to obtain sufficient nutrients causes malnutrition. Nutritional science is the study of nutrition, though it typically emphasizes human nutrition. The type of organism determines what nutrients it needs and how it obtains them. Organisms obtain nutrients by consuming organic matter, consuming inorganic matter, absorbing light, or some combination of these. Some can produce nutrients internally by consuming basic elements, while some must consume other organisms to obtain preexisting nutrients. All forms of life require carbon, energy, and water as well as various other molecules. Animals require complex nutrients such as carbohydrates, lipids, and proteins, obtaining them by consuming other organisms. Humans have developed agriculture and cooking to replace for ...
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Transcription Coregulators
In molecular biology and genetics, transcription coregulators are proteins that interact with transcription factors to either activate or repress the transcription of specific genes. Transcription coregulators that activate gene transcription are referred to as coactivators while those that repress are known as corepressors. The mechanism of action of transcription coregulators is to modify chromatin structure and thereby make the associated DNA more or less accessible to transcription. In humans several dozen to several hundred coregulators are known, depending on the level of confidence with which the characterisation of a protein as a coregulator can be made. One class of transcription coregulators modifies chromatin structure through covalent modification of histones. A second ATP dependent class modifies the conformation of chromatin. Histone acetyltransferases Nuclear DNA is normally tightly wrapped around histones rendering the DNA inaccessible to the general transcr ...
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Lac Operon
The ''lactose'' operon (''lac'' operon) is an operon required for the transport and metabolism of lactose in ''E. coli'' and many other enteric bacteria. Although glucose is the preferred carbon source for most bacteria, the ''lac'' operon allows for the effective digestion of lactose when glucose is not available through the activity of beta-galactosidase. Gene regulation of the ''lac'' operon was the first genetic regulatory mechanism to be understood clearly, so it has become a foremost example of prokaryotic gene regulation. It is often discussed in introductory molecular and cellular biology classes for this reason. This lactose metabolism system was used by François Jacob and Jacques Monod to determine how a biological cell knows which enzyme to synthesize. Their work on the ''lac'' operon won them the Nobel Prize in Physiology in 1965. Bacterial operons are polycistronic transcripts that are able to produce multiple proteins from one mRNA transcript. In this case, when l ...
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