Polysome Profiling
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Polysome Profiling
Polysome profiling is a technique in molecular biology that is used to study the association of mRNAs with ribosomes. It is important to note that this technique is different from ribosome profiling. Both techniques have been reviewed and both are used in analysis of the translatome, but the data they generate are at very different levels of specificity. When employed by experts, the technique is remarkably reproducible: the 3 profiles in the first image are from 3 different experiments. The procedure The procedure begins by making a cell lysate of the cells of interest. This lysate contains polysomes, monosomes (composed of one ribosome residing on an mRNA), the small (40S in eukaryotes) and large (60S in eukaryotes) ribosomal subunits, "free" mRNA and a host of other soluble cellular components. The procedure continues by making a continuous sucrose gradient of continuously variable density in a centrifuge tube. At the concentrations used (15-45% in the example), sucrose does no ...
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Polysomes
A polyribosome (or polysome or ergosome) is a group of ribosomes bound to an Messenger RNA, mRNA molecule like “beads” on a “thread”. It consists of a complex of an mRNA molecule and two or more ribosomes that act to translation (biology), translate mRNA instructions into polypeptides. Originally coined "ergosomes" in 1963, they were further characterized by Jonathan Warner, Paul M. Knopf, and Alex Rich. Polysomes are formed during the Eukaryotic_transcription#Elongation, elongation phase when ribosomes and elongation factors synthesize the encoded polypeptide. Multiple ribosomes move along the coding region of mRNA, creating a polysome. The ability of multiple ribosomes to function on an mRNA molecule explains the limited abundance of mRNA in the cell. Polyribosome structure differs between prokaryotic polysomes, eukaryotic polysomes, and membrane bound polysomes. Polysome activity can be used to measure the level of gene expression through a technique called polysomal pr ...
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Centrifuge
A centrifuge is a device that uses centrifugal force to separate various components of a fluid. This is achieved by spinning the fluid at high speed within a container, thereby separating fluids of different densities (e.g. cream from milk) or liquids from solids. It works by causing denser substances and particles to move outward in the radial direction. At the same time, objects that are less dense are displaced and moved to the centre. In a laboratory centrifuge that uses sample tubes, the radial acceleration causes denser particles to settle to the bottom of the tube, while low-density substances rise to the top. A centrifuge can be a very effective filter that separates contaminants from the main body of fluid. Industrial scale centrifuges are commonly used in manufacturing and waste processing to sediment suspended solids, or to separate immiscible liquids. An example is the cream separator found in dairies. Very high speed centrifuges and ultracentrifuges able to provi ...
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Peptide
Peptides (, ) are short chains of amino acids linked by peptide bonds. Long chains of amino acids are called proteins. Chains of fewer than twenty amino acids are called oligopeptides, and include dipeptides, tripeptides, and tetrapeptides. A polypeptide is a longer, continuous, unbranched peptide chain. Hence, peptides fall under the broad chemical classes of biological polymers and oligomers, alongside nucleic acids, oligosaccharides, polysaccharides, and others. A polypeptide that contains more than approximately 50 amino acids is known as a protein. Proteins consist of one or more polypeptides arranged in a biologically functional way, often bound to ligands such as coenzymes and cofactors, or to another protein or other macromolecule such as DNA or RNA, or to complex macromolecular assemblies. Amino acids that have been incorporated into peptides are termed residues. A water molecule is released during formation of each amide bond.. All peptides except cyclic pep ...
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Chaperone (protein)
In molecular biology, molecular chaperones are proteins that assist the conformational folding or unfolding of large proteins or macromolecular protein complexes. There are a number of classes of molecular chaperones, all of which function to assist large proteins in proper protein folding during or after synthesis, and after partial denaturation. Chaperones are also involved in the translocation of proteins for proteolysis. The first molecular chaperones discovered were a type of assembly chaperones which assist in the assembly of nucleosomes from folded histones and DNA. One major function of molecular chaperones is to prevent the aggregation of misfolded proteins, thus many chaperone proteins are classified as heat shock proteins, as the tendency for protein aggregation is increased by heat stress. The majority of molecular chaperones do not convey any steric information for protein folding, and instead assist in protein folding by binding to and stabilizing folding interme ...
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Svedberg
A Svedberg unit or svedberg (symbol S, sometimes Sv) is a non- SI metric unit for sedimentation coefficients. The Svedberg unit offers a measure of a particle's size indirectly based on its sedimentation rate under acceleration (i.e. how fast a particle of given size and shape settles to the bottom of a solution). The svedberg is a measure of time, defined as exactly 10−13 seconds (100  fs). For biological macromolecules and cell organelles like ribosomes, the sedimentation rate is typically measured as the rate of travel in a centrifuge tube subjected to high g-force. The svedberg (S) is distinct from the SI unit sievert or the non-SI unit sverdrup, which also use the symbol Sv. Naming The unit is named after the Swedish chemist Theodor Svedberg (1884–1971), winner of the 1926 Nobel Prize in chemistry for his work on disperse systems, colloids and his invention of the ultracentrifuge. Factors The Svedberg coefficient is a nonlinear function. A particle's mass, dens ...
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Force Of Gravity
In physics, gravity () is a fundamental interaction which causes mutual attraction between all things with mass or energy. Gravity is, by far, the weakest of the four fundamental interactions, approximately 1038 times weaker than the strong interaction, 1036 times weaker than the electromagnetic force and 1029 times weaker than the weak interaction. As a result, it has no significant influence at the level of subatomic particles. However, gravity is the most significant interaction between objects at the macroscopic scale, and it determines the motion of planets, stars, galaxies, and even light. On Earth, gravity gives weight to physical objects, and the Moon's gravity is responsible for sublunar tides in the oceans (the corresponding antipodal tide is caused by the inertia of the Earth and Moon orbiting one another). Gravity also has many important biological functions, helping to guide the growth of plants through the process of gravitropism and influencing the circulatio ...
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Acceleration
In mechanics, acceleration is the rate of change of the velocity of an object with respect to time. Accelerations are vector quantities (in that they have magnitude and direction). The orientation of an object's acceleration is given by the orientation of the ''net'' force acting on that object. The magnitude of an object's acceleration, as described by Newton's Second Law, is the combined effect of two causes: * the net balance of all external forces acting onto that object — magnitude is directly proportional to this net resulting force; * that object's mass, depending on the materials out of which it is made — magnitude is inversely proportional to the object's mass. The SI unit for acceleration is metre per second squared (, \mathrm). For example, when a vehicle starts from a standstill (zero velocity, in an inertial frame of reference) and travels in a straight line at increasing speeds, it is accelerating in the direction of travel. If the vehicle turns, an acc ...
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Optical Density
Absorbance is defined as "the logarithm of the ratio of incident to transmitted radiant power through a sample (excluding the effects on cell walls)". Alternatively, for samples which scatter light, absorbance may be defined as "the negative logarithm of one minus absorptance, as measured on a uniform sample". The term is used in many technical areas to quantify the results of an experimental measurement. While the term has its origin in quantifying the absorption of light, it is often entangled with quantification of light which is “lost” to a detector system through other mechanisms. What these uses of the term tend to have in common is that they refer to a logarithm of the ratio of a quantity of light incident on a sample or material to that which is detected after the light has interacted with the sample.   The term absorption refers to the physical process of absorbing light, while absorbance does not always measure only absorption; it may measure attenuation (of trans ...
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Density
Density (volumetric mass density or specific mass) is the substance's mass per unit of volume. The symbol most often used for density is ''ρ'' (the lower case Greek letter rho), although the Latin letter ''D'' can also be used. Mathematically, density is defined as mass divided by volume: : \rho = \frac where ''ρ'' is the density, ''m'' is the mass, and ''V'' is the volume. In some cases (for instance, in the United States oil and gas industry), density is loosely defined as its weight per unit volume, although this is scientifically inaccurate – this quantity is more specifically called specific weight. For a pure substance the density has the same numerical value as its mass concentration. Different materials usually have different densities, and density may be relevant to buoyancy, purity and packaging. Osmium and iridium are the densest known elements at standard conditions for temperature and pressure. To simplify comparisons of density across different s ...
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Sucrose
Sucrose, a disaccharide, is a sugar composed of glucose and fructose subunits. It is produced naturally in plants and is the main constituent of white sugar. It has the molecular formula . For human consumption, sucrose is extracted and refined from either sugarcane or sugar beet. Sugar mills – typically located in tropical regions near where sugarcane is grown – crush the cane and produce raw sugar which is shipped to other factories for refining into pure sucrose. Sugar beet factories are located in temperate climates where the beet is grown, and process the beets directly into refined sugar. The sugar-refining process involves washing the raw sugar crystals before dissolving them into a sugar syrup which is filtered and then passed over carbon to remove any residual colour. The sugar syrup is then concentrated by boiling under a vacuum and crystallized as the final purification process to produce crystals of pure sucrose that are clear, odorless, and sweet. Suga ...
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Molecular Biology
Molecular biology is the branch of biology that seeks to understand the molecular basis of biological activity in and between cells, including biomolecular synthesis, modification, mechanisms, and interactions. The study of chemical and physical structure of biological macromolecules is known as molecular biology. Molecular biology was first described as an approach focused on the underpinnings of biological phenomena - uncovering the structures of biological molecules as well as their interactions, and how these interactions explain observations of classical biology. In 1945 the term molecular biology was used by physicist William Astbury. In 1953 Francis Crick, James Watson, Rosalind Franklin, and colleagues, working at Medical Research Council unit, Cavendish laboratory, Cambridge (now the MRC Laboratory of Molecular Biology), made a double helix model of DNA which changed the entire research scenario. They proposed the DNA structure based on previous research done by Ro ...
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