Characterizing a Lambda Red Recombinase Induced Presumptive
... acids of the lacI gene, and replaced the rest of the gene with a linear double-stranded DNA PCR product that confers kanamycin resistance. The PCR products encode a kanamycin cassette with lacI flanking region. The initial step was to transform wild type E.coli C29 with plasmid pKD46 using electropo ...
... acids of the lacI gene, and replaced the rest of the gene with a linear double-stranded DNA PCR product that confers kanamycin resistance. The PCR products encode a kanamycin cassette with lacI flanking region. The initial step was to transform wild type E.coli C29 with plasmid pKD46 using electropo ...
Genetic Transformation of Poinsettia (Euphórbia
... In nature, A. tumefaciens will infect dicotyledonous plants to produce crown-gall disease by transferring genes coding for crown-gall into the plant (Hoekema et al. 1983; Stachel & Nester 1986). This is done by a virulence region (vir-region) located in a tumour inducing plasmid (Tiplasmid) in the A ...
... In nature, A. tumefaciens will infect dicotyledonous plants to produce crown-gall disease by transferring genes coding for crown-gall into the plant (Hoekema et al. 1983; Stachel & Nester 1986). This is done by a virulence region (vir-region) located in a tumour inducing plasmid (Tiplasmid) in the A ...
Rapid Selection of Multiple Gene Integrant for the Production of
... G418 (2 and 4 mg/mL) was near the end of chromosome. These results imply that the multiple gene integration that is necessary for the cell to be resistant to high concentration of G418 always occurred near the chromosomal end. The highest copy number of integration was roughly estimated about 10 by ...
... G418 (2 and 4 mg/mL) was near the end of chromosome. These results imply that the multiple gene integration that is necessary for the cell to be resistant to high concentration of G418 always occurred near the chromosomal end. The highest copy number of integration was roughly estimated about 10 by ...
A dominant mutation in the gene for the Nag
... (White, 1968). The nagCl protein is apparently insensitive to this inducer and can bind to the nag operators and repress in the presence of high levels of GlcNAc &phosphate present in the nagA strains. The insensitivity of the mutant repressor to GlcNAc 6phosphate was confirmed by an in vitro test ( ...
... (White, 1968). The nagCl protein is apparently insensitive to this inducer and can bind to the nag operators and repress in the presence of high levels of GlcNAc &phosphate present in the nagA strains. The insensitivity of the mutant repressor to GlcNAc 6phosphate was confirmed by an in vitro test ( ...
Molecular Cloning, Sequencing, and Expression of the Glutamine
... ginIl was used as a probe to clone the ginIl gene from a size-selected KpnI library of Frankia strain CpIl DNA. The region corresponding to the Frankia sp. strain CpIl glnIl gene was sequenced, and the amino acid sequence was compared with that of the GS gene from the pea and ginIl from B. japonicum ...
... ginIl was used as a probe to clone the ginIl gene from a size-selected KpnI library of Frankia strain CpIl DNA. The region corresponding to the Frankia sp. strain CpIl glnIl gene was sequenced, and the amino acid sequence was compared with that of the GS gene from the pea and ginIl from B. japonicum ...
Physiological characterization of natural transformation in
... is encoded by the vector part of pAVA213-8 and will only be retained when the entire plasmid is integrated into the chromosome. Comparing the number of kanamycin-resistant transformants with kanamycin- and ampicillin-resistant transformants within one transformation experiment (Table 2) showed that ...
... is encoded by the vector part of pAVA213-8 and will only be retained when the entire plasmid is integrated into the chromosome. Comparing the number of kanamycin-resistant transformants with kanamycin- and ampicillin-resistant transformants within one transformation experiment (Table 2) showed that ...
Ch12_lecture - Dr. Brahmbhatt`s Class Handouts
... Biotechnology is any use or alteration of organisms, cells, or biological molecules to achieve specific practical goals. • Today, selective breeding is an important part of biotechnology. • Modern biotechnology frequently uses genetic engineering. • This term refers to direct methods for modifying ...
... Biotechnology is any use or alteration of organisms, cells, or biological molecules to achieve specific practical goals. • Today, selective breeding is an important part of biotechnology. • Modern biotechnology frequently uses genetic engineering. • This term refers to direct methods for modifying ...
Bacterial Transformation: Unlocking the Mysteries of Genetic Material
... Biotechnology is one of the newest and fastest growing scientific fields that has led to many new products routinely used in our day to day lives. The simplest definition of biotechnology is “applied biology”, which means the use of scientific techniques and knowledge and applying it to the developm ...
... Biotechnology is one of the newest and fastest growing scientific fields that has led to many new products routinely used in our day to day lives. The simplest definition of biotechnology is “applied biology”, which means the use of scientific techniques and knowledge and applying it to the developm ...
The serC-aroA operon of Escherichia coli
... frame (ORF) which extends from the Clal site to a point 70 bp before the aroA coding region startpoint. This, together with the observation that insertion of the Clal fragment to the left of the Clal site has no effect on aroA expression, suggested that aroA was part of an operon and was co-ordinate ...
... frame (ORF) which extends from the Clal site to a point 70 bp before the aroA coding region startpoint. This, together with the observation that insertion of the Clal fragment to the left of the Clal site has no effect on aroA expression, suggested that aroA was part of an operon and was co-ordinate ...
Human, yeast and hybrid 3-phosphoglycerate kinase gene
... produced by units c and e on high copy number plasmids in yeast. IFN-ol DNA was used as a common probe. Lanes 1-4 were also analyzed with yPGK DNA as a probe to show that loading of mRNA is essentially identical (see part B of lanes 1-4). Loading is measured by the intensity of the 1500 nucleotide c ...
... produced by units c and e on high copy number plasmids in yeast. IFN-ol DNA was used as a common probe. Lanes 1-4 were also analyzed with yPGK DNA as a probe to show that loading of mRNA is essentially identical (see part B of lanes 1-4). Loading is measured by the intensity of the 1500 nucleotide c ...
W0=2, a stable aneuploid derivative of Candida
... (Scherer & Magee, 1990). One is its lack of a sexual cycle; another is its diploid nature. The first precludes the use of classical genetic analysis to study the properties related to virulence and pathogenicity. The second makes molecular genetics much more cumbersome, since mutants made by classic ...
... (Scherer & Magee, 1990). One is its lack of a sexual cycle; another is its diploid nature. The first precludes the use of classical genetic analysis to study the properties related to virulence and pathogenicity. The second makes molecular genetics much more cumbersome, since mutants made by classic ...
Ammonium transport in Escherichia coli: localization and nucleotide
... Bacterial strains and growth conditions. Strain A52653 (amtA1: :TnlO Tc') was grown in M9 minimal medium, with sodium glutamate (20 mM) as sole source of nitrogen, as described previously (Jayakumar et al., 1986). In complementation assays, restoration of the Amtphenotype to A52653 was monitored on ...
... Bacterial strains and growth conditions. Strain A52653 (amtA1: :TnlO Tc') was grown in M9 minimal medium, with sodium glutamate (20 mM) as sole source of nitrogen, as described previously (Jayakumar et al., 1986). In complementation assays, restoration of the Amtphenotype to A52653 was monitored on ...
Primary Sequence of Ovomucoid Messenger RNA as Determined
... clones, which have been shown to contain no intervening sequences in the 5' noncoding region, are in complete agreement with the data from the cDNA clone. The sequence of the 3' noncoding region was reported by Buell et al. (7). Our initial data, derived from experiments on only one strand in the 3' ...
... clones, which have been shown to contain no intervening sequences in the 5' noncoding region, are in complete agreement with the data from the cDNA clone. The sequence of the 3' noncoding region was reported by Buell et al. (7). Our initial data, derived from experiments on only one strand in the 3' ...
First report of a tetracycline-inducible gene
... animals and plants, in which they may cause severe diseases. Due to their small genome sizes and limited metabolic pathways, they are commonly described as the simplest self-replicating organisms (Peterson & Fraser, 2001; Sirand-Pugnet et al., 2007). Genomes of mollicutes have been among the very fi ...
... animals and plants, in which they may cause severe diseases. Due to their small genome sizes and limited metabolic pathways, they are commonly described as the simplest self-replicating organisms (Peterson & Fraser, 2001; Sirand-Pugnet et al., 2007). Genomes of mollicutes have been among the very fi ...
Primary Sequence of Ovomucoid Messenger RNA
... clones, which have been shown to contain no intervening sequences in the 5' noncoding region, are in complete agreement with the data from the cDNA clone. The sequence of the 3' noncoding region was reported by Buell et al. (7). Our initial data, derived from experiments on only one strand in the 3' ...
... clones, which have been shown to contain no intervening sequences in the 5' noncoding region, are in complete agreement with the data from the cDNA clone. The sequence of the 3' noncoding region was reported by Buell et al. (7). Our initial data, derived from experiments on only one strand in the 3' ...
Cloning of genes from genomic DNA: Part 3
... by PCR were simply a blunt-ended fragment, we could just cut the plasmid with a restriction enzyme that leaves blunt ends as well and ligate the two together (blunt-ended cloning works, but is less efficient than cloning sticky ends). When PCR was first used to clone fragments, many people tried thi ...
... by PCR were simply a blunt-ended fragment, we could just cut the plasmid with a restriction enzyme that leaves blunt ends as well and ligate the two together (blunt-ended cloning works, but is less efficient than cloning sticky ends). When PCR was first used to clone fragments, many people tried thi ...
Chapter 12 Recombinant DNA Technology Key Concepts
... Chapter 14. However, the plasmids that are routinely used as vectors are those that carry genes for drug resistance. The drug-resistance genes are useful because the drug-resistant phenotype can be used to select not only for cells transformed by plasmids, but also for vectors containing recombinant ...
... Chapter 14. However, the plasmids that are routinely used as vectors are those that carry genes for drug resistance. The drug-resistance genes are useful because the drug-resistant phenotype can be used to select not only for cells transformed by plasmids, but also for vectors containing recombinant ...
FischerSpr09
... Based on the assumption that the morphogenetic pathway is the same in all fungi, knocking out the homologous gene in a more amenable fungi (in this case S. cerevisiae) and replacing it with one similar from the target organism (C. albicans) should bring about the same morphogenetic changes. Therefor ...
... Based on the assumption that the morphogenetic pathway is the same in all fungi, knocking out the homologous gene in a more amenable fungi (in this case S. cerevisiae) and replacing it with one similar from the target organism (C. albicans) should bring about the same morphogenetic changes. Therefor ...
Skeletal muscle actin mRNA. Characterization of the 3
... inserts that were sequenced are indicated by dotted lines. Restriction enzymes that were used and the position of their cleavage sites are indicated. The alignment of the cDNA inserts of plasmids p749 and pl50 with actin mRNA was made by comparing the amino acid sequences deduced from the nucleotide ...
... inserts that were sequenced are indicated by dotted lines. Restriction enzymes that were used and the position of their cleavage sites are indicated. The alignment of the cDNA inserts of plasmids p749 and pl50 with actin mRNA was made by comparing the amino acid sequences deduced from the nucleotide ...
An Introduction to Genetic Analysis Chapter 20 Transposable
... mutated to a1, an allele for the colorless phenotype; at another locus, a dominant allele Dt (Dotted) had appeared. The effect of Dt was to produce pigmented dots in the otherwise colorless phenotype of a1/a1 (Figure 20-1). Thus, the original line was very probably A1/A1 ; dt/dt, and the mutations g ...
... mutated to a1, an allele for the colorless phenotype; at another locus, a dominant allele Dt (Dotted) had appeared. The effect of Dt was to produce pigmented dots in the otherwise colorless phenotype of a1/a1 (Figure 20-1). Thus, the original line was very probably A1/A1 ; dt/dt, and the mutations g ...
mschi
... two disadvantages that could be prohibitive during process scaleup (Leonard et al., 2007, 2008; Miyahisa et al., 2005). The first main shortcoming is that fermentation protocols often require two separate cultivation steps to achieve high flavonoid titers. Typically, strains are first grown in rich med ...
... two disadvantages that could be prohibitive during process scaleup (Leonard et al., 2007, 2008; Miyahisa et al., 2005). The first main shortcoming is that fermentation protocols often require two separate cultivation steps to achieve high flavonoid titers. Typically, strains are first grown in rich med ...
IS Elements
... between different DNA molecules. • Conjugative plasmids can move transposons that contain genes for antibiotic resistance from one bacterial cell to another. ...
... between different DNA molecules. • Conjugative plasmids can move transposons that contain genes for antibiotic resistance from one bacterial cell to another. ...
Cloning and functional characterization of temperature responsive
... Another reason of this potential in drought tolerance could be due to its high stomatal control enabling plants to minimize the transpiration rate by keeping also a high level of net CO2 assimilation under water stress conditions (Severino et al., 2012). These physiologic ...
... Another reason of this potential in drought tolerance could be due to its high stomatal control enabling plants to minimize the transpiration rate by keeping also a high level of net CO2 assimilation under water stress conditions (Severino et al., 2012). These physiologic ...
pcr (polymerase chain reaction)
... PCR reaction below. If the PCR product size from a colony matches the size from the positive control, then there is a good chance you have the plasmid of interest. RFP Circuit Teams If you had glowing colonies on your #1A plate with IPTG, then perform colony PCR on 5 glowing colonies you have, one n ...
... PCR reaction below. If the PCR product size from a colony matches the size from the positive control, then there is a good chance you have the plasmid of interest. RFP Circuit Teams If you had glowing colonies on your #1A plate with IPTG, then perform colony PCR on 5 glowing colonies you have, one n ...
Author`s personal copy
... If a highly efficient method for mutant construction is developed, a genome-wide analysis of gene function is possible (Hammelmann and Soppa, 2008). It would not only accelerate the genome-wide identification of the genes responsible for PHA biosynthesis, but may also be used to genetically engineer ...
... If a highly efficient method for mutant construction is developed, a genome-wide analysis of gene function is possible (Hammelmann and Soppa, 2008). It would not only accelerate the genome-wide identification of the genes responsible for PHA biosynthesis, but may also be used to genetically engineer ...
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.