E.coli
... Gel soaked in base to denature duplexes pH readjusted to neutral Sheet of absorbent material placed atop the gel Salt solution is drawn across the gel, perp to the electrophoretic direction, in various ways to carry the DNA onto the sheet Sheet is dried in an oven to tightly attach the DNA to it Inc ...
... Gel soaked in base to denature duplexes pH readjusted to neutral Sheet of absorbent material placed atop the gel Salt solution is drawn across the gel, perp to the electrophoretic direction, in various ways to carry the DNA onto the sheet Sheet is dried in an oven to tightly attach the DNA to it Inc ...
Chromosome-encoded gene cluster for the
... a multi-component aniline dioxygenase and a LysR-type regulator, respectively, while the others (tadD1C1D2C2EFGIJKL) were expected to encode meta-cleavage pathway enzymes for catechol degradation. In addition, it was found that the gene cluster is surrounded by two IS1071 sequences, indicating that ...
... a multi-component aniline dioxygenase and a LysR-type regulator, respectively, while the others (tadD1C1D2C2EFGIJKL) were expected to encode meta-cleavage pathway enzymes for catechol degradation. In addition, it was found that the gene cluster is surrounded by two IS1071 sequences, indicating that ...
Restriction Enzyme digestion of DNA
... fragments will equal the number of restriction sites. Eukaryotic (linear) DNA • If you have one restriction site for an enzyme, you would have 2 fragments, and if you have 2 restriction sites for an enzyme, you would have 3 fragments. In Eukaryotic DNA, the number of fragments is always going to hav ...
... fragments will equal the number of restriction sites. Eukaryotic (linear) DNA • If you have one restriction site for an enzyme, you would have 2 fragments, and if you have 2 restriction sites for an enzyme, you would have 3 fragments. In Eukaryotic DNA, the number of fragments is always going to hav ...
Genetic tools for manipulating Acinetobacter baumannii genome: an
... as a temperature-sensitive replicon. The vector must also carry a selectable marker, usually an antibiotic resistance gene. Sometimes integration vectors also contain oriT so that the construct can be transferred to other bacteria by conjugation. This is especially helpful when the other transfer me ...
... as a temperature-sensitive replicon. The vector must also carry a selectable marker, usually an antibiotic resistance gene. Sometimes integration vectors also contain oriT so that the construct can be transferred to other bacteria by conjugation. This is especially helpful when the other transfer me ...
A physical map of the genome of Hmmophilus
... Digestion of DNA in agarose blocks. Usually digests were carried out on the DNA contained in one-third of a complete plug. Restriction einzyme buffers were diffused into the agarose blocks as outlined below. Plugs or portions of plugs were washed in Eppendorf tubes with 500 1.11 vlolumesof buffer (u ...
... Digestion of DNA in agarose blocks. Usually digests were carried out on the DNA contained in one-third of a complete plug. Restriction einzyme buffers were diffused into the agarose blocks as outlined below. Plugs or portions of plugs were washed in Eppendorf tubes with 500 1.11 vlolumesof buffer (u ...
hybrid DNA molecules
... Such molecules replicate autonomously with an average copy number of 5-10 covalently closed circles per yeast cell and also replicate as a chromosomally integrated structure. This DNA may be physically isolated in intact form from either yeast or E. coli and used to transform either organism at high ...
... Such molecules replicate autonomously with an average copy number of 5-10 covalently closed circles per yeast cell and also replicate as a chromosomally integrated structure. This DNA may be physically isolated in intact form from either yeast or E. coli and used to transform either organism at high ...
Major Contributing Factor in Increased Antibiotic Resistance
... are not readily treatable with antibiotics. In fact, biofilms are resistant to antibiotic levels 10- to 1,000-fold higher than planktonic, or free-floating, bacteria (2). The properties of biofilms resulting in their increased resistance to antibiotics are not thoroughly understood. The close contac ...
... are not readily treatable with antibiotics. In fact, biofilms are resistant to antibiotic levels 10- to 1,000-fold higher than planktonic, or free-floating, bacteria (2). The properties of biofilms resulting in their increased resistance to antibiotics are not thoroughly understood. The close contac ...
Gel electrophoresis of restriction digest
... seen on both closed circular and linear duplex DNA. The amount of DNA to be loaded is an important factor in gel resolution. Between 5 and 200 ng of a single DNA fragment can be loaded into a well. 5ng approaches the minimal amount of an individual DNA fragment that can be detected by ethidium bromi ...
... seen on both closed circular and linear duplex DNA. The amount of DNA to be loaded is an important factor in gel resolution. Between 5 and 200 ng of a single DNA fragment can be loaded into a well. 5ng approaches the minimal amount of an individual DNA fragment that can be detected by ethidium bromi ...
Optimizing bacterial expression and purifica- Biomedical laboratory science,
... Genetic recombination takes place in nature both in animals and plants, where both parents of an individual are the original sources of the DNA, which recombine during meiosis. This recombinant DNA differs only from the parental DNA in the combination of the alleles it contains, but the sequence of ...
... Genetic recombination takes place in nature both in animals and plants, where both parents of an individual are the original sources of the DNA, which recombine during meiosis. This recombinant DNA differs only from the parental DNA in the combination of the alleles it contains, but the sequence of ...
Biotechnology Explorer - Bio-Rad
... them to glow a brilliant green color under ultraviolet light. In this activity, students will learn about the process of moving genes from one organism to another with the aid of a plasmid. In addition to one large chromosome, bacteria naturally contain one or more small circular pieces of DNA calle ...
... them to glow a brilliant green color under ultraviolet light. In this activity, students will learn about the process of moving genes from one organism to another with the aid of a plasmid. In addition to one large chromosome, bacteria naturally contain one or more small circular pieces of DNA calle ...
Biotechnology Explorer - Bio-Rad
... them to glow a brilliant green color under ultraviolet light. In this activity, students will learn about the process of moving genes from one organism to another with the aid of a plasmid. In addition to one large chromosome, bacteria naturally contain one or more small circular pieces of DNA calle ...
... them to glow a brilliant green color under ultraviolet light. In this activity, students will learn about the process of moving genes from one organism to another with the aid of a plasmid. In addition to one large chromosome, bacteria naturally contain one or more small circular pieces of DNA calle ...
Chapter 6 Pichia pastoris
... 1993). The yeast, Pichia pastoris, was selected and developed as an alternative yeast expression host, since it displays a lower degree of glycosylation of the recombinant protein than that associated with expression in S. cerevisiae (Invitrogen Corporation, 2001). It also contain methanol-regulated ...
... 1993). The yeast, Pichia pastoris, was selected and developed as an alternative yeast expression host, since it displays a lower degree of glycosylation of the recombinant protein than that associated with expression in S. cerevisiae (Invitrogen Corporation, 2001). It also contain methanol-regulated ...
90718 Internal v2 3.6 A2 Generic 2006
... has been found to be better that using yeast as the E coli based clone libraries are much more stable which is important when you are trying to accurately identify the sequence of bases in the inserted fragment. ...
... has been found to be better that using yeast as the E coli based clone libraries are much more stable which is important when you are trying to accurately identify the sequence of bases in the inserted fragment. ...
An assessment of the risks associated with the
... the plant cell where the natural integration properties of the T-DNA insert the entire gene ensemble into the chromosomal DNA.6,9–11 Engineered plant DNA can also be delivered into plant cells by particle bombardment (biolistic transformation), when the DNA is literally shot into plant cells on DNA- ...
... the plant cell where the natural integration properties of the T-DNA insert the entire gene ensemble into the chromosomal DNA.6,9–11 Engineered plant DNA can also be delivered into plant cells by particle bombardment (biolistic transformation), when the DNA is literally shot into plant cells on DNA- ...
περισσότερες πληροφορίες
... century, led to the production of human recombinant insulin and revolutionized the treatment of diabetes: • i) Restriction enzymes: In 1962 Werner Arber, a Swiss biochemist, and his colleagues proved the existence of what he called “molecular scissors”, i.e. proteins capable of cutting DNA. • They s ...
... century, led to the production of human recombinant insulin and revolutionized the treatment of diabetes: • i) Restriction enzymes: In 1962 Werner Arber, a Swiss biochemist, and his colleagues proved the existence of what he called “molecular scissors”, i.e. proteins capable of cutting DNA. • They s ...
Biochemistry - Stryer - Science and Technology
... sequence complete genomes: first, small genomes from viruses; then, larger genomes from bacteria; and, finally, eukaryotic genomes, including the 3-billion-base-pair human genome. Scientists are just beginning to exploit the enormous information content of these genome sequences. Finally, recombinan ...
... sequence complete genomes: first, small genomes from viruses; then, larger genomes from bacteria; and, finally, eukaryotic genomes, including the 3-billion-base-pair human genome. Scientists are just beginning to exploit the enormous information content of these genome sequences. Finally, recombinan ...
Temperature-sensitive control of protein activity by conditionally
... expressed by the ptc-Gal4 driver, a striking loss of both wg and sen at the intersection of the a/p and d/v boundaries (arrows in Fig. 3f–h,j–l) is observed at both 29 °C and 18 °C. Mild temperature sensitivity of ptc-Gal4 decreased the activity of dnN at the lower temperature (compare Fig. 3f with ...
... expressed by the ptc-Gal4 driver, a striking loss of both wg and sen at the intersection of the a/p and d/v boundaries (arrows in Fig. 3f–h,j–l) is observed at both 29 °C and 18 °C. Mild temperature sensitivity of ptc-Gal4 decreased the activity of dnN at the lower temperature (compare Fig. 3f with ...
223/AP08 - EDVOTEK
... GFP and its related fluorescent proteins have become an essential tool in cell and molecular biology. Using DNA cloning strategies, proteins can be “tagged” with fluorescent proteins and then expressed in cells. These tags simplify purification because a GFP-labeled protein can be tracked using UV l ...
... GFP and its related fluorescent proteins have become an essential tool in cell and molecular biology. Using DNA cloning strategies, proteins can be “tagged” with fluorescent proteins and then expressed in cells. These tags simplify purification because a GFP-labeled protein can be tracked using UV l ...
Chpt3_Isolating_analyzing_genes.doc
... set of genes in a genome, requires that they be isolated and then studied intensively. Once a gene is “in hand”, in principal one can determine both its biochemical structures and its function(s) in an organism. One of the goals of biochemistry and molecular genetics is to assign particular function ...
... set of genes in a genome, requires that they be isolated and then studied intensively. Once a gene is “in hand”, in principal one can determine both its biochemical structures and its function(s) in an organism. One of the goals of biochemistry and molecular genetics is to assign particular function ...
artificial yeast chromosomes
... in an adenine-deficient minimal medium environment, which makes the yeast dependent on the artificial chromosome for adenine, ensuring that the artificial chromosome is not lost before the start of the experiment. During the experiment, the yeast are grown on YPD, which has plentiful adenine; the ye ...
... in an adenine-deficient minimal medium environment, which makes the yeast dependent on the artificial chromosome for adenine, ensuring that the artificial chromosome is not lost before the start of the experiment. During the experiment, the yeast are grown on YPD, which has plentiful adenine; the ye ...
Bacteroides mobilizable and conjugative genetic elements
... Since CTnDOT appeared to integrate site selectively, the comparions of the sequences of the ends of CTnDOT with the sequences of the integration sites revealed some sequence similarity. One candidate for recognition sequence is a 10-bp sequence that is immediately adjacent to the site where CTnDOT e ...
... Since CTnDOT appeared to integrate site selectively, the comparions of the sequences of the ends of CTnDOT with the sequences of the integration sites revealed some sequence similarity. One candidate for recognition sequence is a 10-bp sequence that is immediately adjacent to the site where CTnDOT e ...
Identification and removal of colanic acid from plasmid DNA
... solvents such as ethanol (EtOH) and by polyethylene glycol. Because polysaccharides are anionic, they co-purify with DNA by anion-exchange chromatography. The commercially available laboratory scale purification methods, including Qiagen, use anionexchange chromatography for at least one major step ...
... solvents such as ethanol (EtOH) and by polyethylene glycol. Because polysaccharides are anionic, they co-purify with DNA by anion-exchange chromatography. The commercially available laboratory scale purification methods, including Qiagen, use anionexchange chromatography for at least one major step ...
K -Channel Transgenes Reduce K Currents in Paramecium
... unexpected. K⫹ channels normally pass outward currents that repolarize the membrane after depolarization. Therefore, the overexpression of K⫹ channels is expected to terminate rather than to prolong membrane depolarization (excitation). Intact PAK11 ORF in the transgene is required for the effect: I ...
... unexpected. K⫹ channels normally pass outward currents that repolarize the membrane after depolarization. Therefore, the overexpression of K⫹ channels is expected to terminate rather than to prolong membrane depolarization (excitation). Intact PAK11 ORF in the transgene is required for the effect: I ...
Teacher`s guide - National Centre for Biotechnology Education
... family. Restriction enzymes are used to cut the DNA into fragments and these are separated by gel electrophoresis. The DNA is then stained so that it can be seen and the class results are combined and analysed. This mirrors the way in which genetic analysis has traditionally been carried out. The sc ...
... family. Restriction enzymes are used to cut the DNA into fragments and these are separated by gel electrophoresis. The DNA is then stained so that it can be seen and the class results are combined and analysed. This mirrors the way in which genetic analysis has traditionally been carried out. The sc ...
Plasmid
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A plasmid is a small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently. They are most commonly found in bacteria as small, circular, double-stranded DNA molecules; however, plasmids are sometimes present in archaea and eukaryotic organisms. In nature, plasmids often carry genes that may benefit the survival of the organism, for example antibiotic resistance. While the chromosomes are big and contain all the essential information for living, plasmids usually are very small and contain only additional information. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms.Plasmids are considered replicons, a unit of DNA capable of replicating autonomously within a suitable host. However, plasmids, like viruses, are not generally classified as life. Plasmids can be transmitted from one bacterium to another (even of another species) via three main mechanisms: transformation, transduction, and conjugation. This host-to-host transfer of genetic material is called horizontal gene transfer, and plasmids can be considered part of the mobilome. Unlike viruses (which encase their genetic material in a protective protein coat called a capsid), plasmids are ""naked"" DNA and do not encode genes necessary to encase the genetic material for transfer to a new host. However, some classes of plasmids encode the conjugative ""sex"" pilus necessary for their own transfer. The size of the plasmid varies from 1 to over 200 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances.The relationship between microbes and plasmid DNA is neither parasitic nor mutualistic, because each implies the presence of an independent species living in a detrimental or commensal state with the host organism. Rather, plasmids provide a mechanism for horizontal gene transfer within a population of microbes and typically provide a selective advantage under a given environmental state. Plasmids may carry genes that provide resistance to naturally occurring antibiotics in a competitive environmental niche, or the proteins produced may act as toxins under similar circumstances, or allow the organism to utilize particular organic compounds that would be advantageous when nutrients are scarce.