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Calf-intestinal Alkaline Phosphatase
Calf-intestinal alkaline phosphatase (CIAP/CIP) is a type of alkaline phosphatase that catalyzes the removal of phosphate groups from the 5' end of DNA strands and phosphomonoesters from RNA. This enzyme is frequently used in DNA sub-cloning, as DNA fragments that lack the 5' phosphate groups cannot ligate. This prevents recircularization of the linearized DNA vector Vector most often refers to: * Euclidean vector, a quantity with a magnitude and a direction * Disease vector, an agent that carries and transmits an infectious pathogen into another living organism Vector may also refer to: Mathematics a ... and improves the yield of the vector containing the appropriate insert. Applications Calf-intestinal alkaline phosphatase can serve as an effective tool for removing uranium from groundwater and soil that can pose major health risks. Furthermore, the toxicity of lipopolysaccharide (LPS) was mitigated by calf-intestinal alkaline phosphatase in mice and piglets, wh ...
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Alkaline Phosphatase
The enzyme alkaline phosphatase (ALP, alkaline phenyl phosphatase, also abbreviated PhoA) is a phosphatase with the physiological role of dephosphorylating compounds. The enzyme is found across a multitude of organisms, prokaryotes and eukaryotes alike, with the same general function, but in different structural forms suitable to the environment they function in. Alkaline phosphatase is found in the periplasmic space of '' E. coli'' bacteria. This enzyme is heat stable and has its maximum activity at high pH. In humans, it is found in many forms depending on its origin within the body – it plays an integral role in metabolism within the liver and development within the skeleton. Due to its widespread prevalence in these areas, its concentration in the bloodstream is used by diagnosticians as a biomarker in helping determine diagnoses such as hepatitis or osteomalacia. The level of alkaline phosphatase in the blood is checked through the ALP test, which is often par ...
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Phosphate Group
Phosphates are the naturally occurring form of the element phosphorus. In chemistry, a phosphate is an anion, salt, functional group or ester derived from a phosphoric acid. It most commonly means orthophosphate, a derivative of orthophosphoric acid, phosphoric acid . The phosphate or orthophosphate ion is derived from phosphoric acid by the removal of three protons . Removal of one proton gives the dihydrogen phosphate ion while removal of two protons gives the hydrogen phosphate ion . These names are also used for salts of those anions, such as ammonium dihydrogen phosphate and trisodium phosphate. File:3-phosphoric-acid-3D-balls.png, Phosphoricacid File:2-dihydrogenphosphate-3D-balls.png, Dihydrogenphosphate File:1-hydrogenphosphate-3D-balls.png, Hydrogenphosphate File:0-phosphate-3D-balls.png, Phosphate or orthophosphate In organic chemistry, phosphate or orthophosphate is an organophosphate, an ester of orthophosphoric acid of the form where one or mo ...
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Phosphomonoesters
Phosphomonoesters (or phosphoric esters) are chemical compounds containing one ester bond and a phosphate group. In biology, phosphomonoesters are needed as the building blocks for the synthesis of Phospholipid cellular membranes, especially those found on neurons. Enzymes which cleave these bonds are known as phosphomonoesterases, or phosphatases. See also * Phosphoric acid * Phospholipid Phospholipids are a class of lipids whose molecule has a hydrophilic "head" containing a phosphate group and two hydrophobic "tails" derived from fatty acids, joined by an alcohol residue (usually a glycerol molecule). Marine phospholipids typ ... References Organophosphates {{ester-stub ...
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Sub-cloning
In molecular biology, subcloning is a technique used to move a particular DNA sequence from a ''parent vector'' to a ''destination vector''. Subcloning is not to be confused with molecular cloning Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their DNA replication, replication within Host (biology), host organisms. The use of the word ''cloning'' re ..., a related technique. Procedure Restriction enzymes are used to excise the gene of interest (the ''insert'') from the parent. The insert is purified in order to isolate it from other DNA molecules. A common purification method is gel isolation. The number of copies of the gene is then amplified using polymerase chain reaction (PCR). Simultaneously, the same restriction enzymes are used to digest (cut) the destination. The idea behind using the same restriction enzymes is to create Complementary DNA, complementary sticky ends, whi ...
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Ligation (molecular Biology)
Ligation is the joining of two nucleotides, or two nucleic acid fragments, into a single polymeric chain through the action of an enzyme known as a ligase. The reaction involves the formation of a phosphodiester bond between the 3'-hydroxyl terminus of one nucleotide and the 5'-phosphoryl terminus of another nucleotide, which results in the two nucleotides being linked consecutively on a single strand. Ligation works in fundamentally the same way for both DNA and RNA. A cofactor is generally involved in the reaction, usually Adenosine triphosphate, ATP or Nicotinamide riboside, NAD+. Eukaryotic ligases belong to the ATP type, while the NAD+ type are found in bacteria (e.g. Escherichia coli, ''E. coli''). Ligation occurs naturally as part of numerous cellular processes, including DNA replication, transcription, splicing, and recombination, and is also an essential laboratory procedure in molecular cloning, whereby DNA fragments are joined to create recombinant DNA molecules (such a ...
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Vector (molecular Biology)
In molecular cloning, a vector is any particle (e.g., plasmids, cosmids, Lambda phages) used as a vehicle to artificially carry a foreign nucleic acid sequence, nucleic sequence – usually DNA – into another Cell (biology), cell, where it can DNA replication, be replicated and/or Gene expression, expressed. A vector containing foreign DNA is termed recombinant DNA. The four major types of vectors are plasmids, viral vectors, cosmids, and Bacterial artificial chromosome, artificial chromosomes. Of these, the most commonly used vectors are plasmids. Common to all engineered vectors are an origin of replication, a multiple cloning site, multicloning site, and a selectable marker. The vector itself generally carries a DNA sequence that consists of an Insert (molecular biology), insert (in this case the transgene) and a larger sequence that serves as the "backbone" of the vector. The purpose of a vector which transfers genetic information to another cell is typically to isolate, mu ...
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Enzymes
An enzyme () is a protein that acts as a biological catalyst by accelerating chemical reactions. The molecules upon which enzymes may act are called substrates, and the enzyme converts the substrates into different molecules known as products. Almost all metabolic processes in the cell need enzyme catalysis in order to occur at rates fast enough to sustain life. Metabolic pathways depend upon enzymes to catalyze individual steps. The study of enzymes is called ''enzymology'' and the field of pseudoenzyme analysis recognizes that during evolution, some enzymes have lost the ability to carry out biological catalysis, which is often reflected in their amino acid sequences and unusual 'pseudocatalytic' properties. Enzymes are known to catalyze more than 5,000 biochemical reaction types. Other biocatalysts include catalytic RNA molecules, also called ribozymes. They are sometimes described as a ''type'' of enzyme rather than being ''like'' an enzyme, but even in the d ...
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