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Clone (genetics)
Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word ''cloning'' refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine. In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest, then treated with enzymes in the test tube to generate smaller DNA fragments. Subsequently, these fragments are then combined with vector DNA to generate recombinant DNA molecules. The recombinant DNA is then ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Steps Of Molecular Cloning
Step(s) or STEP may refer to: Common meanings * Steps, making a staircase * Walking * Dance move * Military step, or march ** Marching Arts Films and television * ''Steps'' (TV series), Hong Kong * ''Step'' (film), US, 2017 Literature * ''Steps'' (novel), by Jerzy Kosinski * Systematic Training for Effective Parenting, a book series Music * Step (music), pitch change * Steps (pop group), UK * ''Step'' (Kara album), 2011, South Korea ** "Step" (Kara song) * ''Step'' (Meg album), 2007, Japan * "Step" (Vampire Weekend song) * "Step" (ClariS song) Organizations * STEP (company), Belgium * Society of Trust and Estate Practitioners, international professional body for advisers who specialise in inheritance and succession planning * Board on Science, Technology, and Economic Policy of the U.S. National Academies * Solving the E-waste Problem, a UN organization Science, technology, and mathematics * Step (software), a physics simulator in KDE * Step function, in mathema ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Blue White Screen
Blue is one of the three primary colours in the RYB colour model (traditional colour theory), as well as in the RGB (additive) colour model. It lies between violet and cyan on the spectrum of visible light. The term ''blue'' generally describes colours perceived by humans observing light with a dominant wavelength that's between approximately 450 and 495 nanometres. Most blues contain a slight mixture of other colours; azure contains some green, while ultramarine contains some violet. The clear daytime sky and the deep sea appear blue because of an optical effect known as Rayleigh scattering. An optical effect called the Tyndall effect explains blue eyes. Distant objects appear more blue because of another optical effect called aerial perspective. Blue has been an important colour in art and decoration since ancient times. The semi-precious stone lapis lazuli was used in ancient Egypt for jewellery and ornament and later, in the Renaissance, to make the pigment ultr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
EcoRI
EcoRI (pronounced "eco R one") is a restriction endonuclease enzyme isolated from species '' E. coli.'' It is a restriction enzyme that cleaves DNA double helices into fragments at specific sites, and is also a part of the restriction modification system. The ''Eco'' part of the enzyme's name originates from the species from which it was isolated"E" denotes generic name, "Escherichia", and "co" denotes species name, "coli"while the R represents the particular strain, in this case RY13, and the I denotes that it was the first enzyme isolated from this strain. In molecular biology it is used as a restriction enzyme. EcoRI creates 4 nucleotide sticky ends with 5' end overhangs of AATT. The nucleic acid recognition sequence where the enzyme cuts is G↓AATTC, which has a palindromic complementary sequence of CTTAA↓G. Other restriction enzymes, depending on their cut sites, can also leave 3' overhangs or blunt ends with no overhangs. History EcoRI is an example of type II restrict ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
DNA End
DNA ends refer to the properties of the ends of linear DNA molecules, which in molecular biology are described as "sticky" or "blunt" based on the shape of the complementary strands at the terminus. In sticky ends, one strand is longer than the other (typically by at least a few nucleotides), such that the longer strand has bases which are left unpaired. In blunt ends, both strands are of equal length – i.e. they end at the same base position, leaving no unpaired bases on either strand. The concept is used in molecular biology, in cloning, or when subcloning insert DNA into vector DNA. Such ends may be generated by restriction enzymes that break the molecule's phosphodiester backbone at specific locations, which themselves belong to a larger class of enzymes called exonucleases and endonucleases. A restriction enzyme that cuts the backbones of both strands at non-adjacent locations leaves a staggered cut, generating two overlapping sticky ends, while an enzyme that makes a str ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
BamHI2
BamHI (pronounced "Bam H one") (from '' Bacillus amyloliquefaciens'') is a type II restriction endonuclease, having the capacity for recognizing short sequences (6 bp) of DNA and specifically cleaving them at a target site. This exhibit focuses on the structure-function relations of BamHI as described by Newman, et al. (1995). BamHI binds at the recognition sequence 5'-GGATCC-3', and cleaves these sequences just after the 5'-guanine on each strand. This cleavage results in sticky ends which are 4 bp long. In its unbound form, BamHI displays a central b sheet, which resides in between α-helices. BamHI undergoes a series of unconventional conformational changes upon DNA recognition. This allows the DNA to maintain its normal B-DNA conformation without distorting to facilitate enzyme binding. BamHI is a symmetric dimer. DNA is bound in a large cleft that is formed between dimers; the enzyme binds in a "crossover" manner. Each BamHI subunit makes the majority of its backbone c ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Subcloning
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, 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 sticky ends, which will facilitate ligation later on. A phosphatase, commonly calf-intestinal alkaline phosphatase (CIAP), is also added to prevent self-ligation of the destination vector. The digested destination vector is isolated/purified. The insert and the destination vector ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Shuttle Vector
A shuttle vector is a vector (usually a plasmid) constructed so that it can propagate in two different host species. Therefore, DNA inserted into a shuttle vector can be tested or manipulated in two different cell types. The main advantage of these vectors is they can be manipulated in ''E. coli'', then used in a system which is more difficult or slower to use (e.g. yeast). Shuttle vectors include plasmids that can propagate in eukaryotes and prokaryotes (e.g. both ''Saccharomyces cerevisiae'' and ''Escherichia coli'') or in different species of bacteria (e.g. both ''E. coli'' and ''Rhodococcus erythropolis''). There are also adenovirus shuttle vectors, which can propagate in ''E. coli'' and mammals. Shuttle vectors are frequently used to quickly make multiple copies of the gene in ''E. coli'' (amplification). They can also be used for ''in vitro'' experiments and modifications (e.g. mutagenesis, PCR). One of the most common types of shuttle vectors is the yeast shuttle vector. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Expression Vector
An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector (molecular biology), vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein Genetic code, encoded by the gene. Expression vectors are the basic tools in biotechnology for the protein production, production of proteins. The Vector (molecular biology), vector is engineered to contain regulatory sequences that act as Enhancer (genetics), enhancer and Promoter (biology), promoter regions and lead to efficient transcription of the gene carried on the expression vector. The goal of a well-designed expression vector is the efficient production of protein, and this may be achieved by the production of significant amount of stable messenger RNA, which can then be Translation (biology), translated into protein. The expression of a protein may be tightly ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Yeast Artificial Chromosome
Yeast artificial chromosomes (YACs) are genetically engineered chromosomes derived from the DNA of the yeast, ''Saccharomyces cerevisiae' which is then ligated into a bacterial plasmid. By inserting large fragments of DNA, from 100–1000 kb, the inserted sequences can be cloned and physically mapped using a process called chromosome walking. This is the process that was initially used for the Human Genome Project, however due to stability issues, YACs were abandoned for the use of bacterial artificial chromosomebr> The bakers' yeast ''S. cerevisiae'' is one of the most important experimental organisms for studying eukaryotic molecular genetics. Beginning with the initial research of the Rankin et al., Strul et al., and Hsaio et al., the inherently fragile chromosome was stabilized by discovering the necessary autonomously replicating sequence (ARS); a refined YAC utilizing this data was described in 1983 by Murray et al. The primary components of a YAC are the ARS, centr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Bacterial Artificial Chromosome
A bacterial artificial chromosome (BAC) is a DNA construct, based on a functional fertility plasmid (or F-plasmid), used for transforming and cloning in bacteria, usually '' E. coli''. F-plasmids play a crucial role because they contain partition genes that promote the even distribution of plasmids after bacterial cell division. The bacterial artificial chromosome's usual insert size is 150–350 kbp. A similar cloning vector called a PAC has also been produced from the DNA of P1 bacteriophage. BACs were often used to sequence the genomes of organisms in genome projects, for example the Human Genome Project, though they have been replaced by more modern technologies. In BAC sequencing, short piece of the organism's DNA is amplified as an insert in BACs, and then sequenced. Finally, the sequenced parts are rearranged '' in silico'', resulting in the genomic sequence of the organism. BACs were replaced with faster and less laborious sequencing methods like whole genome shot ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Plasmid Vector
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 using ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
