Cloning and expression of the phosphotriesterase
... fusion when cultures were grown in the absence of phosphate, suggesting that it might be part of the Pho regulon, but the phosphate-regulated promoter was not cloned in this study. This is believed to be the first study in which a gene required for an organism to grow with OP pesticides as a phospho ...
... fusion when cultures were grown in the absence of phosphate, suggesting that it might be part of the Pho regulon, but the phosphate-regulated promoter was not cloned in this study. This is believed to be the first study in which a gene required for an organism to grow with OP pesticides as a phospho ...
Microbiology
... hsdR marker with the wild-type dnaC gene. One of these isolates was named strain KW1. The physical absence of the gus operon in strain KW1 was confirmed by Southern hybridization analysis using a 6.2 kb EcoRI-Hind111 fragment from pRAJ289 containing gusABC as a probe. Strain KW1 was used as the reci ...
... hsdR marker with the wild-type dnaC gene. One of these isolates was named strain KW1. The physical absence of the gus operon in strain KW1 was confirmed by Southern hybridization analysis using a 6.2 kb EcoRI-Hind111 fragment from pRAJ289 containing gusABC as a probe. Strain KW1 was used as the reci ...
Recombinant DNA technology - The Centre for Plant Sciences
... them. NcoI is very popular, because it spans an ATG sequence, and is often engineered at the beginning of the coding region. Yes, your guess was right, it wasn’t an accident, it is done on purpose and when you look at genes in nature, they don’t always have this restriction site in this position, on ...
... them. NcoI is very popular, because it spans an ATG sequence, and is often engineered at the beginning of the coding region. Yes, your guess was right, it wasn’t an accident, it is done on purpose and when you look at genes in nature, they don’t always have this restriction site in this position, on ...
Study on the Genetic Transformation of Gentian by Gene Recombinant
... inducing rol gene express and producing hair root. Meanwhile, using the Agrobacterium Rhizogenes R1000 infect gentian directly as a comparison, we built two sets of transform system of Agrobacterium for hairy root through researching on all kinds of factors deeply and optimizing transform condition. ...
... inducing rol gene express and producing hair root. Meanwhile, using the Agrobacterium Rhizogenes R1000 infect gentian directly as a comparison, we built two sets of transform system of Agrobacterium for hairy root through researching on all kinds of factors deeply and optimizing transform condition. ...
Principles of Nucleic Acid Separation by Agarose Gel Electrophoresis
... 3. Use insulated gloves to transfer the flask into a water bath at 55°C. When the melted gel has cooled, add ethidium bromide to a final concentration of 0.5 μg/ml. Mix the gel solution thoroughly by gentle swirling. 4. While the agarose solution is cooling, choose an appropriate comb for forming th ...
... 3. Use insulated gloves to transfer the flask into a water bath at 55°C. When the melted gel has cooled, add ethidium bromide to a final concentration of 0.5 μg/ml. Mix the gel solution thoroughly by gentle swirling. 4. While the agarose solution is cooling, choose an appropriate comb for forming th ...
DNA cloning
... eral different enzymes. If the sticky end contains a 5¢ overhang, then any one of several different DNA polymerases can be used to add the missing bases to the 3¢ OH using the 5¢ overhang as a template. A 3¢ overhang cannot be filled in, rather the overhang must be removed. Many DNA polymerases have ...
... eral different enzymes. If the sticky end contains a 5¢ overhang, then any one of several different DNA polymerases can be used to add the missing bases to the 3¢ OH using the 5¢ overhang as a template. A 3¢ overhang cannot be filled in, rather the overhang must be removed. Many DNA polymerases have ...
Structural organization of the transfer RNA gene clusters of cholera
... fragment. The sizes of the various fragments were obtained from their relative mobilities on gel with those of λ DNA-HindIII fragments. 2.7 Southern blot analysis Briefly, phage φ 149 DNA was digested to completion with various restriction enzymes, singly or with respective double combinations, and ...
... fragment. The sizes of the various fragments were obtained from their relative mobilities on gel with those of λ DNA-HindIII fragments. 2.7 Southern blot analysis Briefly, phage φ 149 DNA was digested to completion with various restriction enzymes, singly or with respective double combinations, and ...
Application to Modify an Approved Exempt or Notifiable Low Risk
... (b) must not code for a toxin with an LD50 of less than 100 g/kg; and (c) must not code for a toxin with an LD50 of 100 g/kg or more, if the intention is to express the toxin at high levels; and (d) must not be uncharacterised nucleic acid from a toxin-producing organism; and (e) must not include ...
... (b) must not code for a toxin with an LD50 of less than 100 g/kg; and (c) must not code for a toxin with an LD50 of 100 g/kg or more, if the intention is to express the toxin at high levels; and (d) must not be uncharacterised nucleic acid from a toxin-producing organism; and (e) must not include ...
Multiple Domains Exist within the Upstream Activator
... and sugar derivatives called opines. The inciting strain of Agrobacterium can utilize these opines as a carbon and sometimes a nitrogen source. Agrobacterium strains can be classified based upon their ability to catabolize particular opines. (For recent reviews of the crown gall tumorigenesis proces ...
... and sugar derivatives called opines. The inciting strain of Agrobacterium can utilize these opines as a carbon and sometimes a nitrogen source. Agrobacterium strains can be classified based upon their ability to catabolize particular opines. (For recent reviews of the crown gall tumorigenesis proces ...
Lesson Overview - Dr. Thornton`s Courses
... Recently it has become possible to eliminate particular genes by constructing DNA molecules with two ends that will sometimes recombine with specific sequences in the host chromosome. Once they recombine, the host gene normally found between those two sequences may be lost or specifically replaced w ...
... Recently it has become possible to eliminate particular genes by constructing DNA molecules with two ends that will sometimes recombine with specific sequences in the host chromosome. Once they recombine, the host gene normally found between those two sequences may be lost or specifically replaced w ...
Applied and Environmental Microbiology
... n the last decade, much attention has been given to betaproteobacteria in the genus Burkholderia mainly because of their great metabolic versatility, their capacity for colonizing very distinct niches, and their ability to cause serious infections in humans (1). Currently the Burkholderia genus comp ...
... n the last decade, much attention has been given to betaproteobacteria in the genus Burkholderia mainly because of their great metabolic versatility, their capacity for colonizing very distinct niches, and their ability to cause serious infections in humans (1). Currently the Burkholderia genus comp ...
Quick Ligation™ Kit
... work well. For optimum ligation, the volume of DNA and insert should be 10 µl before adding 2X Quick Ligation Buffer. For DNA volumes greater than 10 µl, increase the volume of 2X Quick Ligation Buffer such that it remains 50% of the reaction and correspondingly increase the volume of ligase. The ov ...
... work well. For optimum ligation, the volume of DNA and insert should be 10 µl before adding 2X Quick Ligation Buffer. For DNA volumes greater than 10 µl, increase the volume of 2X Quick Ligation Buffer such that it remains 50% of the reaction and correspondingly increase the volume of ligase. The ov ...
The first true obligately syntrophic propionate
... were inoculated with 0?2 g freeze-dried granular sludge. After addition of 20 mM sodium propionate and 10 % (v/v) of a H2/CO2-grown culture of Methanospirillum hungatei JF-1T, the enrichment culture was incubated at 37 uC. After 3 months, the propionate was depleted and CH4 was produced. Acetate was ...
... were inoculated with 0?2 g freeze-dried granular sludge. After addition of 20 mM sodium propionate and 10 % (v/v) of a H2/CO2-grown culture of Methanospirillum hungatei JF-1T, the enrichment culture was incubated at 37 uC. After 3 months, the propionate was depleted and CH4 was produced. Acetate was ...
Amino acid alterations essential for increasing the catalytic activity of
... acrylamide (5%) gel electrophoresis of DNA were performed 206-257 of EII). pUCl2-V was constructed from a 2736-bp EcoRI - NaeI fragment of pUC12-8PS and a 250-bp EcoRI as described previously [9]. NaeI fragment of pUC12-7PS (residues 185-257 of EII’). For construction of plasmids pHK13 - 17, the fol ...
... acrylamide (5%) gel electrophoresis of DNA were performed 206-257 of EII). pUCl2-V was constructed from a 2736-bp EcoRI - NaeI fragment of pUC12-8PS and a 250-bp EcoRI as described previously [9]. NaeI fragment of pUC12-7PS (residues 185-257 of EII’). For construction of plasmids pHK13 - 17, the fol ...
Manipulating the Plasmodium Genome
... replicate as unrearranged monomeric units, with an average copy number of 15 per nucleus (van Dijk et al., 1997). These plasmids appear to be fairly stable, even in the absence of drug pressure. In P. falciparum, however, the situation is more complex. Plasmids rapidly form large concatemers (Kadeko ...
... replicate as unrearranged monomeric units, with an average copy number of 15 per nucleus (van Dijk et al., 1997). These plasmids appear to be fairly stable, even in the absence of drug pressure. In P. falciparum, however, the situation is more complex. Plasmids rapidly form large concatemers (Kadeko ...
20Sexual Reproduction, Meiosis, and Genetic Recombination
... Because sexual reproduction combines genetic information from two different parents into a single offspring, at some point in its life cycle every sexually reproducing organism has cells that contain two copies of each type of chromosome, one inherited from each parent. The two members of each chrom ...
... Because sexual reproduction combines genetic information from two different parents into a single offspring, at some point in its life cycle every sexually reproducing organism has cells that contain two copies of each type of chromosome, one inherited from each parent. The two members of each chrom ...
Adaptation of Sucrose Metabolism in the Escherichia coli Wild
... genes might have been transferred relatively recently to the E. coli wild-type EC3132 at around the time when the different strains of the enteric bacteria diverged. We found evidence that a mobile genetic element, which used the gene argW for site-specific integration into the chromosome, was proba ...
... genes might have been transferred relatively recently to the E. coli wild-type EC3132 at around the time when the different strains of the enteric bacteria diverged. We found evidence that a mobile genetic element, which used the gene argW for site-specific integration into the chromosome, was proba ...
The Fly Genome
... site included in it The cos site signals for packaging into l phage, thus the plasmid, including a large insert can be packaged into phage particles without having to waste space on l genes A cosmid combined with a cloned insert cannot exceed the 53 kb limit on how much can be packaged into the l ph ...
... site included in it The cos site signals for packaging into l phage, thus the plasmid, including a large insert can be packaged into phage particles without having to waste space on l genes A cosmid combined with a cloned insert cannot exceed the 53 kb limit on how much can be packaged into the l ph ...
Cloning - Allegiance
... Cloning created identical genes. It is a process of replicating a genetic constitution, thus hampering the diversity in genes. While lessening the diversity in genes, we weaken our ability of adaptation. Cloning is also detrimental to the beauty that lies in diversity. While cloning allows man to ta ...
... Cloning created identical genes. It is a process of replicating a genetic constitution, thus hampering the diversity in genes. While lessening the diversity in genes, we weaken our ability of adaptation. Cloning is also detrimental to the beauty that lies in diversity. While cloning allows man to ta ...
Phospholipid synthesis in Borrelia burgdorferi: BB0249 and BB0721
... protocols (Ames, 1968) and TLC of the extracts was performed. Staining of the TLC plates confirmed that PC and PG are the major phospholipids in the B. burgdorferi membrane (data not shown) and suggested that this method was a reliable method for examining B. burgdorferi membrane phospholipids. We t ...
... protocols (Ames, 1968) and TLC of the extracts was performed. Staining of the TLC plates confirmed that PC and PG are the major phospholipids in the B. burgdorferi membrane (data not shown) and suggested that this method was a reliable method for examining B. burgdorferi membrane phospholipids. We t ...
20. Transposable Genetic Elements
... receptor and the regulator are termed controlling elements. In the examples considered so far, the unstable allele is said to be nonautonomous: it can revert only in the presence of the regulator. Sometimes, however, a system such as the Ac-Ds system can produce an unstable allele that is autonomous ...
... receptor and the regulator are termed controlling elements. In the examples considered so far, the unstable allele is said to be nonautonomous: it can revert only in the presence of the regulator. Sometimes, however, a system such as the Ac-Ds system can produce an unstable allele that is autonomous ...
Foundations of Biology
... bulge appears in images of polytene chromosomes In situ hybridization using cosmid clones mapped to that region show hybridization on the outside of this structure, but not in the middle Figure from http://www.helsinki.fi/~saura/EM/xdiv2.html ...
... bulge appears in images of polytene chromosomes In situ hybridization using cosmid clones mapped to that region show hybridization on the outside of this structure, but not in the middle Figure from http://www.helsinki.fi/~saura/EM/xdiv2.html ...
Insertional inactivation studies of the csmA and csmC genes of the
... smaller HindIII fragments of 0.85-kb and 0.72-kb that arise from the introduced HindIII sites that £ank the 6 cartridge in the insertionally inactivated csmA gene. Even after repeated restreaking and growth on up to 600 Wg ml31 streptomycin (Fig. 1B,C, lanes 4), no segregation of alleles occurred. T ...
... smaller HindIII fragments of 0.85-kb and 0.72-kb that arise from the introduced HindIII sites that £ank the 6 cartridge in the insertionally inactivated csmA gene. Even after repeated restreaking and growth on up to 600 Wg ml31 streptomycin (Fig. 1B,C, lanes 4), no segregation of alleles occurred. T ...
PPT - Bruce Blumberg
... • Separate size range of interest by PFGE (pulsed field gel electrophoresis) • ligate to vector and transform by electroporation • What is the potential flaw for all these methods? BioSci Blumberg lecture 1 ...
... • Separate size range of interest by PFGE (pulsed field gel electrophoresis) • ligate to vector and transform by electroporation • What is the potential flaw for all these methods? BioSci Blumberg lecture 1 ...
Plasmid
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.