Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.
Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.
Mutagenesis where the mutation is caused by the introduction of foreign DNA sequences into a gene or extragenic sequence. This may occur spontaneously in vivo or be experimentally induced in vivo or in vitro. Proviral DNA insertions into or adjacent to a cellular proto-oncogene can interrupt GENETIC TRANSLATION of the coding sequences or interfere with recognition of regulatory elements and cause unregulated expression of the proto-oncogene resulting in tumor formation.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
The naturally occurring or experimentally induced replacement of one or more AMINO ACIDS in a protein with another. If a functionally equivalent amino acid is substituted, the protein may retain wild-type activity. Substitution may also diminish, enhance, or eliminate protein function. Experimentally induced substitution is often used to study enzyme activities and binding site properties.
The relationship between the chemical structure of a compound and its biological or pharmacological activity. Compounds are often classed together because they have structural characteristics in common including shape, size, stereochemical arrangement, and distribution of functional groups.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
Discrete segments of DNA which can excise and reintegrate to another site in the genome. Most are inactive, i.e., have not been found to exist outside the integrated state. DNA transposable elements include bacterial IS (insertion sequence) elements, Tn elements, the maize controlling elements Ac and Ds, Drosophila P, gypsy, and pogo elements, the human Tigger elements and the Tc and mariner elements which are found throughout the animal kingdom.
Proteins prepared by recombinant DNA technology.
Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.
The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.
The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.
Proteins found in any species of bacterium.
The rate dynamics in chemical or physical systems.
The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
A thiol-containing non-essential amino acid that is oxidized to form CYSTINE.
The level of protein structure in which regular hydrogen-bond interactions within contiguous stretches of polypeptide chain give rise to alpha helices, beta strands (which align to form beta sheets) or other types of coils. This is the first folding level of protein conformation.
The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.
The functional hereditary units of BACTERIA.
A mutation caused by the substitution of one nucleotide for another. This results in the DNA molecule having a change in a single base pair.
Chemical agents that increase the rate of genetic mutation by interfering with the function of nucleic acids. A clastogen is a specific mutagen that causes breaks in chromosomes.
A nitrosourea compound with alkylating, carcinogenic, and mutagenic properties.
A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.
The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction.
A non-essential amino acid that occurs in high levels in its free state in plasma. It is produced from pyruvate by transamination. It is involved in sugar and acid metabolism, increases IMMUNITY, and provides energy for muscle tissue, BRAIN, and the CENTRAL NERVOUS SYSTEM.
Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.
Established cell cultures that have the potential to propagate indefinitely.
An error-prone mechanism or set of functions for repairing damaged microbial DNA. SOS functions (a concept reputedly derived from the SOS of the international distress signal) are involved in DNA repair and mutagenesis, in cell division inhibition, in recovery of normal physiological conditions after DNA repair, and possibly in cell death when DNA damage is extensive.
An essential amino acid that is required for the production of HISTAMINE.
A sequence of amino acids in a polypeptide or of nucleotides in DNA or RNA that is similar across multiple species. A known set of conserved sequences is represented by a CONSENSUS SEQUENCE. AMINO ACID MOTIFS are often composed of conserved sequences.
Biochemical identification of mutational changes in a nucleotide sequence.
The region of an enzyme that interacts with its substrate to cause the enzymatic reaction.
Proteins obtained from ESCHERICHIA COLI.
The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
A test used to determine whether or not complementation (compensation in the form of dominance) will occur in a cell with a given mutant phenotype when another mutant genome, encoding the same mutant phenotype, is introduced into that cell.
The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
An antineoplastic agent with alkylating properties. It also acts as a mutagen by damaging DNA and is used experimentally for that effect.
Deoxyribonucleic acid that makes up the genetic material of bacteria.
Deletion of sequences of nucleic acids from the genetic material of an individual.
Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes.
Procedures by which protein structure and function are changed or created in vitro by altering existing or synthesizing new structural genes that direct the synthesis of proteins with sought-after properties. Such procedures may include the design of MOLECULAR MODELS of proteins using COMPUTER GRAPHICS or other molecular modeling techniques; site-specific mutagenesis (MUTAGENESIS, SITE-SPECIFIC) of existing genes; and DIRECTED MOLECULAR EVOLUTION techniques to create new genes.
A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.
An essential amino acid. It is often added to animal feed.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
The techniques used to produce molecules exhibiting properties that conform to the demands of the experimenter. These techniques combine methods of generating structural changes with methods of selection. They are also used to examine proposed mechanisms of evolution under in vitro selection conditions.
One of the non-essential amino acids commonly occurring in the L-form. It is found in animals and plants, especially in sugar cane and sugar beets. It may be a neurotransmitter.
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
Commonly observed structural components of proteins formed by simple combinations of adjacent secondary structures. A commonly observed structure may be composed of a CONSERVED SEQUENCE which can be represented by a CONSENSUS SEQUENCE.
The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat.
Proteins produced from GENES that have acquired MUTATIONS.
DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes.
That portion of the electromagnetic spectrum immediately below the visible range and extending into the x-ray frequencies. The longer wavelengths (near-UV or biotic or vital rays) are necessary for the endogenous synthesis of vitamin D and are also called antirachitic rays; the shorter, ionizing wavelengths (far-UV or abiotic or extravital rays) are viricidal, bactericidal, mutagenic, and carcinogenic and are used as disinfectants.
A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
A non-essential amino acid that is involved in the metabolic control of cell functions in nerve and brain tissue. It is biosynthesized from ASPARTIC ACID and AMMONIA by asparagine synthetase. (From Concise Encyclopedia Biochemistry and Molecular Biology, 3rd ed)
A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.
Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria.
DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.
A non-essential amino acid occurring in natural form as the L-isomer. It is synthesized from GLYCINE or THREONINE. It is involved in the biosynthesis of PURINES; PYRIMIDINES; and other amino acids.
CELL LINES derived from the CV-1 cell line by transformation with a replication origin defective mutant of SV40 VIRUS, which codes for wild type large T antigen (ANTIGENS, POLYOMAVIRUS TRANSFORMING). They are used for transfection and cloning. (The CV-1 cell line was derived from the kidney of an adult male African green monkey (CERCOPITHECUS AETHIOPS).)
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
The reconstruction of a continuous two-stranded DNA molecule without mismatch from a molecule which contained damaged regions. The major repair mechanisms are excision repair, in which defective regions in one strand are excised and resynthesized using the complementary base pairing information in the intact strand; photoreactivation repair, in which the lethal and mutagenic effects of ultraviolet light are eliminated; and post-replication repair, in which the primary lesions are not repaired, but the gaps in one daughter duplex are filled in by incorporation of portions of the other (undamaged) daughter duplex. Excision repair and post-replication repair are sometimes referred to as "dark repair" because they do not require light.
DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair.
An essential amino acid that is physiologically active in the L-form.
A change from planar to elliptic polarization when an initially plane-polarized light wave traverses an optically active medium. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
The process by which two molecules of the same chemical composition form a condensation product or polymer.
A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed)
An essential amino acid that is necessary for normal growth in infants and for NITROGEN balance in adults. It is a precursor of INDOLE ALKALOIDS in plants. It is a precursor of SEROTONIN (hence its use as an antidepressant and sleep aid). It can be a precursor to NIACIN, albeit inefficiently, in mammals.
A genetic rearrangement through loss of segments of DNA or RNA, bringing sequences which are normally separated into close proximity. This deletion may be detected using cytogenetic techniques and can also be inferred from the phenotype, indicating a deletion at one specific locus.
CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells.
A species of CERCOPITHECUS containing three subspecies: C. tantalus, C. pygerythrus, and C. sabeus. They are found in the forests and savannah of Africa. The African green monkey (C. pygerythrus) is the natural host of SIMIAN IMMUNODEFICIENCY VIRUS and is used in AIDS research.
A set of three nucleotides in a protein coding sequence that specifies individual amino acids or a termination signal (CODON, TERMINATOR). Most codons are universal, but some organisms do not produce the transfer RNAs (RNA, TRANSFER) complementary to all codons. These codons are referred to as unassigned codons (CODONS, NONSENSE).
Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.
Tests of chemical substances and physical agents for mutagenic potential. They include microbial, insect, mammalian cell, and whole animal tests.
Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.
In bacteria, a group of metabolically related genes, with a common promoter, whose transcription into a single polycistronic MESSENGER RNA is under the control of an OPERATOR REGION.
The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
The chemical or biochemical addition of carbohydrate or glycosyl groups to other chemicals, especially peptides or proteins. Glycosyl transferases are used in this biochemical reaction.
A large collection of DNA fragments cloned (CLONING, MOLECULAR) from a given organism, tissue, organ, or cell type. It may contain complete genomic sequences (GENOMIC LIBRARY) or complementary DNA sequences, the latter being formed from messenger RNA and lacking intron sequences.
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
Production of new arrangements of DNA by various mechanisms such as assortment and segregation, CROSSING OVER; GENE CONVERSION; GENETIC TRANSFORMATION; GENETIC CONJUGATION; GENETIC TRANSDUCTION; or mixed infection of viruses.
Transport proteins that carry specific substances in the blood or across cell membranes.
Injuries to DNA that introduce deviations from its normal, intact structure and which may, if left unrepaired, result in a MUTATION or a block of DNA REPLICATION. These deviations may be caused by physical or chemical agents and occur by natural or unnatural, introduced circumstances. They include the introduction of illegitimate bases during replication or by deamination or other modification of bases; the loss of a base from the DNA backbone leaving an abasic site; single-strand breaks; double strand breaks; and intrastrand (PYRIMIDINE DIMERS) or interstrand crosslinking. Damage can often be repaired (DNA REPAIR). If the damage is extensive, it can induce APOPTOSIS.
The heritable modification of the properties of a competent bacterium by naked DNA from another source. The uptake of naked DNA is a naturally occuring phenomenon in some bacteria. It is often used as a GENE TRANSFER TECHNIQUE.
The process of cleaving a chemical compound by the addition of a molecule of water.
Change brought about to an organisms genetic composition by unidirectional transfer (TRANSFECTION; TRANSDUCTION, GENETIC; CONJUGATION, GENETIC, etc.) and incorporation of foreign DNA into prokaryotic or eukaryotic cells by recombination of part or all of that DNA into the cell's genome.
A nitrosoguanidine derivative with potent mutagenic and carcinogenic properties.
Directed modification of the gene complement of a living organism by such techniques as altering the DNA, substituting genetic material by means of a virus, transplanting whole nuclei, transplanting cell hybrids, etc.
A low-energy attractive force between hydrogen and another element. It plays a major role in determining the properties of water, proteins, and other compounds.
The facilitation of biochemical reactions with the aid of naturally occurring catalysts such as ENZYMES.
Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.
A theoretical representative nucleotide or amino acid sequence in which each nucleotide or amino acid is the one which occurs most frequently at that site in the different sequences which occur in nature. The phrase also refers to an actual sequence which approximates the theoretical consensus. A known CONSERVED SEQUENCE set is represented by a consensus sequence. Commonly observed supersecondary protein structures (AMINO ACID MOTIFS) are often formed by conserved sequences.
Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.
Any method used for determining the location of and relative distances between genes on a chromosome.
The process by which a DNA molecule is duplicated.
A family of recombinases initially identified in BACTERIA. They catalyze the ATP-driven exchange of DNA strands in GENETIC RECOMBINATION. The product of the reaction consists of a duplex and a displaced single-stranded loop, which has the shape of the letter D and is therefore called a D-loop structure.
A non-essential amino acid. In animals it is synthesized from PHENYLALANINE. It is also the precursor of EPINEPHRINE; THYROID HORMONES; and melanin.
A mutation in which a codon is mutated to one directing the incorporation of a different amino acid. This substitution may result in an inactive or unstable product. (From A Dictionary of Genetics, King & Stansfield, 5th ed)
A non-essential amino acid naturally occurring in the L-form. Glutamic acid is the most common excitatory neurotransmitter in the CENTRAL NERVOUS SYSTEM.
The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.
Any of various enzymatically catalyzed post-translational modifications of PEPTIDES or PROTEINS in the cell of origin. These modifications include carboxylation; HYDROXYLATION; ACETYLATION; PHOSPHORYLATION; METHYLATION; GLYCOSYLATION; ubiquitination; oxidation; proteolysis; and crosslinking and result in changes in molecular weight and electrophoretic motility.
Polymers made up of a few (2-20) nucleotides. In molecular genetics, they refer to a short sequence synthesized to match a region where a mutation is known to occur, and then used as a probe (OLIGONUCLEOTIDE PROBES). (Dorland, 28th ed)
The sequential correspondence of nucleotides in one nucleic acid molecule with those of another nucleic acid molecule. Sequence homology is an indication of the genetic relatedness of different organisms and gene function.
A non-essential amino acid. It is found primarily in gelatin and silk fibroin and used therapeutically as a nutrient. It is also a fast inhibitory neurotransmitter.
The degree of pathogenicity within a group or species of microorganisms or viruses as indicated by case fatality rates and/or the ability of the organism to invade the tissues of the host. The pathogenic capacity of an organism is determined by its VIRULENCE FACTORS.
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
An essential aromatic amino acid that is a precursor of MELANIN; DOPAMINE; noradrenalin (NOREPINEPHRINE), and THYROXINE.
Chemical groups containing the covalent disulfide bonds -S-S-. The sulfur atoms can be bound to inorganic or organic moieties.
Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.
The characteristic 3-dimensional shape and arrangement of multimeric proteins (aggregates of more than one polypeptide chain).
The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.
Enzymes that recombine DNA segments by a process which involves the formation of a synapse between two DNA helices, the cleavage of single strands from each DNA helix and the ligation of a DNA strand from one DNA helix to the other. The resulting DNA structure is called a Holliday junction which can be resolved by DNA REPLICATION or by HOLLIDAY JUNCTION RESOLVASES.
Proteins obtained from the species SACCHAROMYCES CEREVISIAE. The function of specific proteins from this organism are the subject of intense scientific interest and have been used to derive basic understanding of the functioning similar proteins in higher eukaryotes.
Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin.
A type of mutation in which a number of NUCLEOTIDES deleted from or inserted into a protein coding sequence is not divisible by three, thereby causing an alteration in the READING FRAMES of the entire coding sequence downstream of the mutation. These mutations may be induced by certain types of MUTAGENS or may occur spontaneously.
Highly reactive chemicals that introduce alkyl radicals into biologically active molecules and thereby prevent their proper functioning. Many are used as antineoplastic agents, but most are very toxic, with carcinogenic, mutagenic, teratogenic, and immunosuppressant actions. They have also been used as components in poison gases.
A sequence of successive nucleotide triplets that are read as CODONS specifying AMINO ACIDS and begin with an INITIATOR CODON and end with a stop codon (CODON, TERMINATOR).
The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.
Genes whose expression is easily detectable and therefore used to study promoter activity at many positions in a target genome. In recombinant DNA technology, these genes may be attached to a promoter region of interest.
The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.
Proteins found in any species of virus.
The location of the atoms, groups or ions relative to one another in a molecule, as well as the number, type and location of covalent bonds.
An essential amino acid occurring naturally in the L-form, which is the active form. It is found in eggs, milk, gelatin, and other proteins.
Chromosomal, biochemical, intracellular, and other methods used in the study of genetics.
The genetic unit consisting of three structural genes, an operator and a regulatory gene. The regulatory gene controls the synthesis of the three structural genes: BETA-GALACTOSIDASE and beta-galactoside permease (involved with the metabolism of lactose), and beta-thiogalactoside acetyltransferase.
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
Proteins found in any species of fungus.
Any member of the group of ENDOPEPTIDASES containing at the active site a serine residue involved in catalysis.
A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.
Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment.
A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
An enzyme that catalyzes the conversion of 5-phosphoribosyl-1-pyrophosphate and hypoxanthine, guanine, or 6-mercaptopurine to the corresponding 5'-mononucleotides and pyrophosphate. The enzyme is important in purine biosynthesis as well as central nervous system functions. Complete lack of enzyme activity is associated with the LESCH-NYHAN SYNDROME, while partial deficiency results in overproduction of uric acid. EC
The interaction of two or more substrates or ligands with the same binding site. The displacement of one by the other is used in quantitative and selective affinity measurements.
A genus of the family Muridae consisting of eleven species. C. migratorius, the grey or Armenian hamster, and C. griseus, the Chinese hamster, are the two species used in biomedical research.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation.
Proteins which maintain the transcriptional quiescence of specific GENES or OPERONS. Classical repressor proteins are DNA-binding proteins that are normally bound to the OPERATOR REGION of an operon, or the ENHANCER SEQUENCES of a gene until a signal occurs that causes their release.
An essential branched-chain amino acid important for hemoglobin formation.
A non-essential amino acid present abundantly throughout the body and is involved in many metabolic processes. It is synthesized from GLUTAMIC ACID and AMMONIA. It is the principal carrier of NITROGEN in the body and is an important energy source for many cells.
A species of gram-positive bacteria that is a common soil and water saprophyte.
A group of enzymes that catalyzes the hydrolysis of terminal, non-reducing beta-D-galactose residues in beta-galactosides. Deficiency of beta-Galactosidase A1 may cause GANGLIOSIDOSIS, GM1.
Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
Theoretical representations that simulate the behavior or activity of genetic processes or phenomena. They include the use of mathematical equations, computers, and other electronic equipment.
The integration of exogenous DNA into the genome of an organism at sites where its expression can be suitably controlled. This integration occurs as a result of homologous recombination.
The assembly of the QUATERNARY PROTEIN STRUCTURE of multimeric proteins (MULTIPROTEIN COMPLEXES) from their composite PROTEIN SUBUNITS.
A non-essential amino acid that is synthesized from GLUTAMIC ACID. It is an essential component of COLLAGEN and is important for proper functioning of joints and tendons.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
The class of all enzymes catalyzing oxidoreduction reactions. The substrate that is oxidized is regarded as a hydrogen donor. The systematic name is based on donor:acceptor oxidoreductase. The recommended name will be dehydrogenase, wherever this is possible; as an alternative, reductase can be used. Oxidase is only used in cases where O2 is the acceptor. (Enzyme Nomenclature, 1992, p9)
Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.
Spectroscopic method of measuring the magnetic moment of elementary particles such as atomic nuclei, protons or electrons. It is employed in clinical applications such as NMR Tomography (MAGNETIC RESONANCE IMAGING).
A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5.
A rigorously mathematical analysis of energy relationships (heat, work, temperature, and equilibrium). It describes systems whose states are determined by thermal parameters, such as temperature, in addition to mechanical and electromagnetic parameters. (From Hawley's Condensed Chemical Dictionary, 12th ed)
The functional hereditary units of FUNGI.
The commonest and widest ranging species of the clawed "frog" (Xenopus) in Africa. This species is used extensively in research. There is now a significant population in California derived from escaped laboratory animals.
Theoretical representations that simulate the behavior or activity of biological processes or diseases. For disease models in living animals, DISEASE MODELS, ANIMAL is available. Biological models include the use of mathematical equations, computers, and other electronic equipment.
Single chains of amino acids that are the units of multimeric PROTEINS. Multimeric proteins can be composed of identical or non-identical subunits. One or more monomeric subunits may compose a protomer which itself is a subunit structure of a larger assembly.
The degree of 3-dimensional shape similarity between proteins. It can be an indication of distant AMINO ACID SEQUENCE HOMOLOGY and used for rational DRUG DESIGN.
Membrane proteins whose primary function is to facilitate the transport of molecules across a biological membrane. Included in this broad category are proteins involved in active transport (BIOLOGICAL TRANSPORT, ACTIVE), facilitated transport and ION CHANNELS.
A method (first developed by E.M. Southern) for detection of DNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
Genes that have a suppressor allele or suppressor mutation (SUPPRESSION, GENETIC) which cancels the effect of a previous mutation, enabling the wild-type phenotype to be maintained or partially restored. For example, amber suppressors cancel the effect of an AMBER NONSENSE MUTATION.
The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle.
Mutation process that restores the wild-type PHENOTYPE in an organism possessing a mutationally altered GENOTYPE. The second "suppressor" mutation may be on a different gene, on the same gene but located at a distance from the site of the primary mutation, or in extrachromosomal genes (EXTRACHROMOSOMAL INHERITANCE).
An essential branched-chain aliphatic amino acid found in many proteins. It is an isomer of LEUCINE. It is important in hemoglobin synthesis and regulation of blood sugar and energy levels.
Protein modules with conserved ligand-binding surfaces which mediate specific interaction functions in SIGNAL TRANSDUCTION PATHWAYS and the specific BINDING SITES of their cognate protein LIGANDS.
Analysis of PEPTIDES that are generated from the digestion or fragmentation of a protein or mixture of PROTEINS, by ELECTROPHORESIS; CHROMATOGRAPHY; or MASS SPECTROMETRY. The resulting peptide fingerprints are analyzed for a variety of purposes including the identification of the proteins in a sample, GENETIC POLYMORPHISMS, patterns of gene expression, and patterns diagnostic for diseases.
Organic salts or esters of methanesulfonic acid.
Genes whose loss of function or gain of function MUTATION leads to the death of the carrier prior to maturity. They may be essential genes (GENES, ESSENTIAL) required for viability, or genes which cause a block of function of an essential gene at a time when the essential gene function is required for viability.
A representation, generally small in scale, to show the structure, construction, or appearance of something. (From Random House Unabridged Dictionary, 2d ed)
Biologically active DNA which has been formed by the in vitro joining of segments of DNA from different sources. It includes the recombination joint or edge of a heteroduplex region where two recombining DNA molecules are connected.
Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.
The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments.
The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.
A set of genes descended by duplication and variation from some ancestral gene. Such genes may be clustered together on the same chromosome or dispersed on different chromosomes. Examples of multigene families include those that encode the hemoglobins, immunoglobulins, histocompatibility antigens, actins, tubulins, keratins, collagens, heat shock proteins, salivary glue proteins, chorion proteins, cuticle proteins, yolk proteins, and phaseolins, as well as histones, ribosomal RNA, and transfer RNA genes. The latter three are examples of reiterated genes, where hundreds of identical genes are present in a tandem array. (King & Stanfield, A Dictionary of Genetics, 4th ed)
Deoxyribonucleic acid that makes up the genetic material of viruses.
The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.
Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme.
Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.
A branched-chain essential amino acid that has stimulant activity. It promotes muscle growth and tissue repair. It is a precursor in the penicillin biosynthetic pathway.
The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.
Proteins found in the nucleus of a cell. Do not confuse with NUCLEOPROTEINS which are proteins conjugated with nucleic acids, that are not necessarily present in the nucleus.
The accumulation of an electric charge on a object
Actual loss of portion of a chromosome.
The thermodynamic interaction between a substance and WATER.
Processes that stimulate the GENETIC TRANSCRIPTION of a gene or set of genes.
A serotype of Salmonella enterica that is a frequent agent of Salmonella gastroenteritis in humans. It also causes PARATYPHOID FEVER.
Proteins isolated from the outer membrane of Gram-negative bacteria.
Female germ cells derived from OOGONIA and termed OOCYTES when they enter MEIOSIS. The primary oocytes begin meiosis but are arrested at the diplotene state until OVULATION at PUBERTY to give rise to haploid secondary oocytes or ova (OVUM).
A genus of bacteria that form a nonfragmented aerial mycelium. Many species have been identified with some being pathogenic. This genus is responsible for producing a majority of the ANTI-BACTERIAL AGENTS of practical value.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.

Mrj encodes a DnaJ-related co-chaperone that is essential for murine placental development. (1/7552)

We have identified a novel gene in a gene trap screen that encodes a protein related to the DnaJ co-chaperone in E. coli. The gene, named Mrj (mammalian relative of DnaJ) was expressed throughout development in both the embryo and placenta. Within the placenta, expression was particularly high in trophoblast giant cells but moderate levels were also observed in trophoblast cells of the chorion at embryonic day 8.5, and later in the labyrinth which arises from the attachment of the chorion to the allantois (a process called chorioallantoic fusion). Insertion of the ROSAbetageo gene trap vector into the Mrj gene created a null allele. Homozygous Mrj mutants died at mid-gestation due to a failure of chorioallantoic fusion at embryonic day 8.5, which precluded formation of the mature placenta. At embryonic day 8.5, the chorion in mutants was morphologically normal and expressed the cell adhesion molecule beta4 integrin that is known to be required for chorioallantoic fusion. However, expression of the chorionic trophoblast-specific transcription factor genes Err2 and Gcm1 was significantly reduced. The mutants showed no abnormal phenotypes in other trophoblast cell types or in the embryo proper. This study indicates a previously unsuspected role for chaperone proteins in placental development and represents the first genetic analysis of DnaJ-related protein function in higher eukaryotes. Based on a survey of EST databases representing different mouse tissues and embryonic stages, there are 40 or more DnaJ-related genes in mammals. In addition to Mrj, at least two of these genes are also expressed in the developing mouse placenta. The specificity of the developmental defect in Mrj mutants suggests that each of these genes may have unique tissue and cellular activities.  (+info)

Regulation of body length and male tail ray pattern formation of Caenorhabditis elegans by a member of TGF-beta family. (2/7552)

We have identified a new member of the TGF-beta superfamily, CET-1, from Caenorhabditis elegans, which is expressed in the ventral nerve cord and other neurons. cet-1 null mutants have shortened bodies and male tail abnormal phenotype resembling sma mutants, suggesting cet-1, sma-2, sma-3 and sma-4 share a common pathway. Overexpression experiments demonstrated that cet-1 function requires wild-type sma genes. Interestingly, CET-1 appears to affect body length in a dose-dependent manner. Heterozygotes for cet-1 displayed body lengths ranging between null mutant and wild type, and overexpression of CET-1 in wild-type worms elongated body length close to lon mutants. In male sensory ray patterning, lack of cet-1 function results in ray fusions. Epistasis analysis revealed that mab-21 lies downstream and is negatively regulated by the cet-1/sma pathway in the male tail. Our results show that cet-1 controls diverse biological processes during C. elegans development probably through different target genes.  (+info)

Isocitrate lyase of Ashbya gossypii--transcriptional regulation and peroxisomal localization. (3/7552)

The isocitrate lyase-encoding gene AgICL1 from the filamentous hemiascomycete Ashbya gossypii was isolated by heterologous complementation of a Saccharomyces cerevisiae icl1d mutant. The open reading frame of 1680 bp encoded a protein of 560 amino acids with a calculated molecular weight of 62584. Disruption of the AgICL1 gene led to complete loss of AgIcl1p activity and inability to grow on oleic acid as sole carbon source. Compartmentation of AgIcl1p in peroxisomes was demonstrated both by Percoll density gradient centrifugation and by immunogold labeling of ultrathin sections using specific antibodies. This fitted with the peroxisomal targeting signal AKL predicted from the C-terminal DNA sequence. Northern blot analysis with mycelium grown on different carbon sources as well as AgICL1 promoter replacement with the constitutive AgTEF promoter revealed a regulation at the transcriptional level. AgICL1 was subject to glucose repression, derepressed by glycerol, partially induced by the C2 compounds ethanol and acetate, and fully induced by soybean oil.  (+info)

Crystal structure of the FMN-binding domain of human cytochrome P450 reductase at 1.93 A resolution. (4/7552)

The crystal structure of the FMN-binding domain of human NADPH-cytochrome P450 reductase (P450R-FMN), a key component in the cytochrome P450 monooxygenase system, has been determined to 1.93 A resolution and shown to be very similar both to the global fold in solution (Barsukov I et al., 1997, J Biomol NMR 10:63-75) and to the corresponding domain in the 2.6 A crystal structure of intact rat P450R (Wang M et al., 1997, Proc Nat Acad Sci USA 94:8411-8416). The crystal structure of P450R-FMN reported here confirms the overall similarity of its alpha-beta-alpha architecture to that of the bacterial flavodoxins, but reveals differences in the position, number, and length of the helices relative to the central beta-sheet. The marked similarity between P450R-FMN and flavodoxins in the interactions between the FMN and the protein, indicate a striking evolutionary conservation of the FMN binding site. The P450R-FMN molecule has an unusual surface charge distribution, leading to a very strong dipole, which may be involved in docking cytochrome P450 into place for electron transfer near the FMN. Several acidic residues near the FMN are identified by mutagenesis experiments to be important for electron transfer to P4502D6 and to cytochrome c, a clear indication of the part of the molecular surface that is likely to be involved in substrate binding. Somewhat different parts are found to be involved in binding cytochrome P450 and cytochrome c.  (+info)

Tolerance of a protein to multiple polar-to-hydrophobic surface substitutions. (5/7552)

Hydrophobic substitutions at solvent-exposed positions in two alpha-helical regions of the bacteriophage P22 Arc repressor were introduced by combinatorial mutagenesis. In helix A, hydrophobic residues were tolerated individually at each of the five positions examined, but multiple substitutions were poorly tolerated as shown by the finding that mutants with more than two additional hydrophobic residues were biologically inactive. Several inactive helix A variants were purified and found to have reduced thermal stability relative to wild-type Arc, with a rough correlation between the number of polar-to-hydrophobic substitutions and the magnitude of the stability defect. Quite different results were obtained in helix B, where variants with as many as five polar-to-hydrophobic substitutions were found to be biologically active and one variant with three hydrophobic substitutions had a t(m) 6 degrees C higher than wild-type. By contrast, a helix A mutant with three similar polar-to-hydrophobic substitutions was 23 degrees C less stable than wild-type. Also, one set of three polar-to-hydrophobic substitutions in helix B was tolerated when introduced into the wild-type background but not when introduced into an equally active mutant having a nearly identical structure. Context effects occur both when comparing different regions of the same protein and when comparing the same region in two different homologues.  (+info)

Analysis of 4-phosphopantetheinylation of polyhydroxybutyrate synthase from Ralstonia eutropha: generation of beta-alanine auxotrophic Tn5 mutants and cloning of the panD gene region. (6/7552)

The postulated posttranslational modification of the polyhydroxybutyrate (PHA) synthase from Ralstonia eutropha by 4-phosphopantetheine was investigated. Four beta-alanine auxotrophic Tn5-induced mutants of R. eutropha HF39 were isolated, and two insertions were mapped in an open reading frame with strong similarity to the panD gene from Escherichia coli, encoding L-aspartate-1-decarboxylase (EC, whereas two other insertions were mapped in an open reading frame (ORF) with strong similarity to the NAD(P)+ transhydrogenase (EC alpha 1 subunit, encoded by the pntAA gene from Escherichia coli. The panD gene was cloned by complementation of the panD mutant of R. eutropha Q20. DNA sequencing of the panD gene region (3,312 bp) revealed an ORF of 365 bp, encoding a protein with 63 and 67% amino acid sequence similarity to PanD from E. coli and Bacillus subtilis, respectively. Subcloning of only this ORF into vectors pBBR1MCS-3 and pBluescript KS- led to complementation of the panD mutants of R. eutropha and E. coli SJ16, respectively. panD-encoded L-aspartate-1-decarboxylase was further confirmed by an enzymatic assay. Upstream of panD, an ORF with strong similarity to pntAA from E. coli, encoding NAD(P)+ transhydrogenase subunit alpha 1 was found; downstream of panD, two ORFs with strong similarity to pntAB and pntB, encoding subunits alpha 2 and beta of the NAD(P)+ transhydrogenase, respectively, were identified. Thus, a hitherto undetermined organization of pan and pnt genes was found in R. eutropha. Labeling experiments using one of the R. eutropha panD mutants and [2-14C]beta-alanine provided no evidence that R. eutropha PHA synthase is covalently modified by posttranslational attachment of 4-phosphopantetheine, nor did the E. coli panD mutant exhibit detectable labeling of functional PHA synthase from R. eutropha.  (+info)

Molecular characterization of the nitrite-reducing system of Staphylococcus carnosus. (7/7552)

Characterization of a nitrite reductase-negative Staphylococcus carnosus Tn917 mutant led to the identification of the nir operon, which encodes NirBD, the dissimilatory NADH-dependent nitrite reductase; SirA, the putative oxidase and chelatase, and SirB, the uroporphyrinogen III methylase, both of which are necessary for biosynthesis of the siroheme prosthetic group; and NirR, which revealed no convincing similarity to proteins with known functions. We suggest that NirR is essential for nir promoter activity. In the absence of NirR, a weak promoter upstream of sirA seems to drive transcription of sirA, nirB, nirD, and sirB in the stationary-growth phase. In primer extension experiments one predominant and several weaker transcription start sites were identified in the nir promoter region. Northern blot analyses indicated that anaerobiosis and nitrite are induction factors of the nir operon: cells grown aerobically with nitrite revealed small amounts of full-length transcript whereas cells grown anaerobically with or without nitrite showed large amounts of full-length transcript. Although a transcript is detectable, no nitrite reduction occurs in cells grown aerobically with nitrite, indicating an additional oxygen-controlled step at the level of translation, enzyme folding, assembly, or insertion of prosthetic groups. The nitrite-reducing activity expressed during anaerobiosis is switched off reversibly when the oxygen tension increases, most likely due to competition for electrons with the aerobic respiratory chain. Another gene, nirC, is located upstream of the nir operon. nirC encodes a putative integral membrane-spanning protein of unknown function. A nirC mutant showed no distinct phenotype.  (+info)

Protein ProQ influences osmotic activation of compatible solute transporter ProP in Escherichia coli K-12. (8/7552)

ProP is an osmoregulatory compatible solute transporter in Escherichia coli K-12. Mutation proQ220::Tn5 decreased the rate constant for and the extent of ProP activation by an osmotic upshift but did not alter proP transcription or the ProP protein level. Allele proQ220::Tn5 was isolated, and the proQ sequence was determined. Locus proQ is upstream from prc (tsp) at 41.2 centisomes on the genetic map. The proQ220::Tn5 and prc phenotypes were different, however. Gene proQ is predicted to encode a 232-amino-acid, basic, hydrophilic protein (molecular mass, 25,876 Da; calculated isoelectric point, 9.66; 32% D, E, R, or K; 54.5% polar amino acids). The insertion of PCR-amplified proQ into vector pBAD24 produced a plasmid containing the wild-type proQ open reading frame, the expression of which yielded a soluble protein with an apparent molecular mass of 30 kDa. Antibodies raised against the overexpressed ProQ protein detected cross-reactive material in proQ+ bacteria but not in proQ220::Tn5 bacteria. ProQ may be a structural element that influences the osmotic activation of ProP at a posttranslational level.  (+info)

The transformation from ductal carcinoma in situ (DCIS) to invasive breast cancer (IBC) is a crucial step in breast cancer progression. The specific alterations that govern this transition have not been elucidated. HER2/neu is frequently overexpressed in DCIS but is less common in IBC suggesting additional requirements for transformation. To identify genes capable of cooperating with HER2/neu to fully transform mammary epithelial cells, we used an insertional mutagenesis screen on cells isolated from wild-type neu expressing mice and identified the E3 ligase HACE1 as a HER2 cooperative tumor suppressor gene. Loss of HACE1 expression is commonly seen in clinical multiple cancer datasets including breast cancer. HACE1 down-regulation in normal human mammary epithelial cells (HMECs) results in the accumulation of the activated GTP-bound Rac1 partially transforming these cells. Overexpression of HER2 activates Rac1, which further accumulates upon HACE1 loss resulting in Rac1 hyperactivation. While the
Define insertional mutagenesis. insertional mutagenesis synonyms, insertional mutagenesis pronunciation, insertional mutagenesis translation, English dictionary definition of insertional mutagenesis. Noun 1. insertional mutagenesis - a mutation caused by the insertion of exogenous DNA into a genome genetic science, genetics - the branch of biology that...
TY - JOUR. T1 - Insertional mutagenesis by the Tol2 transposon-mediated enhancer trap approach generated mutations in two developmental genes. T2 - tcf7 and synembryn-like. AU - Nagayoshi, Saori. AU - Hayashi, Eriko. AU - Abe, Gembu. AU - Osato, Naoki. AU - Asakawa, Kazuhide. AU - Urasaki, Akihiro. AU - Horikawa, Kazuki. AU - Ikeo, Kazuho. AU - Takeda, Hiroyuki. AU - Kawakami, Koichi. PY - 2008/1. Y1 - 2008/1. N2 - Gene trap and enhancer trap methods using transposon or retrovirus have been recently described in zebrafish. However, insertional mutants using these methods have not been reported. We report here development of an enhancer trap method by using the Tol2 transposable element and identification and characterization of insertional mutants. We created 73 fish lines that carried single copy insertions of an enhancer trap construct, which contained the zebrafish hsp70 promoter and the GFP gene, in their genome and expressed GFP in specific cells, tissues and organs, indicating that the ...
Fast MultiSite Mutagenesis System,Mutagenesis System,Cloning and Mutagenesis System,Products,Beijing TransGen Biotech Co.Ltd,OverviewContents& storageCitations & referencesRelated ImagesDownloadOverviewDescriptionFast MultiSite Mutagenesis Syst
TY - JOUR. T1 - Cc.snf5, a gene encoding a putative component of the SWI/SNF chromatin remodeling complex, is essential for sexual development in the agaricomycete Coprinopsis cinerea. AU - Ando, Yuki. AU - Nakazawa, Takehito. AU - Oka, Kunihiko. AU - Nakahori, Kiyoshi. AU - Kamada, Takashi. PY - 2013/1. Y1 - 2013/1. N2 - We characterized a Coprinopsis cinerea mutant strain, Spe20, defective in fruiting initiation, which was isolated after restriction enzyme-mediated integration (REMI) mutagenesis of a homokaryotic fruiting strain, 326. A plasmid rescue followed by complementation experiments, RACE, and cDNA analyses revealed that the gene, a mutation of which is responsible for the phenotype, is predicted to encode a protein that exhibits a high similarity to yeast Snf5p, a key component of the chromatin remodeling complex SWI/SNF, and named Cc.snf5. Cc.Snf5 is, however, different from Snf5p in that the former has, in addition to an Snf5 domain comprising N-terminal repeat1 (rp1) and C-terminal ...
Tn-Seq is an experimental method for probing the functions of genes through construction of complex random transposon insertion libraries and quantification of each mutants abundance using next-generation sequencing. An important emerging application of Tn-Seq is for identifying genetic interactions, which involves comparing Tn mutant libraries generated in different genetic backgrounds (e.g. wild-type strain versus knockout strain). Several analytical methods have been proposed for analyzing Tn-Seq data to identify genetic interactions, including estimating relative fitness ratios and fitting a generalized linear model. However, these have limitations which necessitate an improved approach. We present a hierarchical Bayesian method for identifying genetic interactions through quantifying the statistical significance of changes in enrichment. The analysis involves a four-way comparison of insertion counts across datasets to identify transposon mutants that differentially affect bacterial fitness
Double transgenic Xenopus laevis embryos were generated using restriction enzyme-mediated integration using SV/Venus/cSCL+19 and gamma-crystallin-mCherry, which was derived from gamma-crystallin-green fluorescent protein (GFP). SV/venus/cSCL+19 enhancer (from Rita Ferreira) and cry-mCherry were co-injected ...
Insertional mutagenesis approaches use oncoretroviruses or transposons to trigger cancer in mice by widespread integration into the cellular genome and activation of oncogenes near the integration site. Mapping the genomic integration sites in tumors allows the identification of genomic regions that are recurrently hit in independent tumors (defined as Common Insertion Sites, CIS) (5), which host genes likely involved in cancer development (6).. We have recently developed a new specifically tailored LV-based insertional mutagen that allowed the induction of hepatocellular carcinoma in 3 different mouse models (4). In order to identify the integration sites from LV-induced tumors and control nontumoral samples, we exploited LAM-PCR (1) followed by 454 pyrosequencing. The analysis of LV integrations in tumors allowed the identification of 4 new liver cancer genes. Here we describe how we applied the LAM-PCR protocol1 for the retrieval of LV integrations from the hepatocellular carcinomas induced ...
Baker, Lindsey A., Tiriac, Hervé, Clevers, Hans, Tuveson, David A. (April 2016) Modeling Pancreatic Cancer with Organoids. Trends in Cancer, 2 (4). pp. 176-190. ISSN 2405-8033 Bapiro, T. E., Frese, K. K., Courtin, A., Bramhall, J. L., Madhu, B., Cook, N., Neesse, A., Griffiths, J. R., Tuveson, D. A., Jodrell, D. I., Richards, F. M. (May 2014) Gemcitabine diphosphate choline is a major metabolite linked to the Kennedy pathway in pancreatic cancer models in vivo. British Journal of Cancer, 111 (2). pp. 318-325. ISSN 0007-0920 Bergerson, R. J., Collier, L. S., Sarver, A. L., Been, R. A., Lugthart, S., Diers, M. D., Zuber, J., Rappaport, A. R., Nixon, M. J., Silverstein, K. A. T., Fan, D. H., Lamblin, A. F. J., Wolff, L., Kersey, J. H., Delwel, R., Lowe, S. W., OSullivan, M. G., Kogan, S. C., Adams, D. J., Largaespada, D. A. (May 2012) An insertional mutagenesis screen identifies genes that cooperate with Mll-AF9 in a murine leukemogenesis model. Blood, 119 (19). pp. 4512-4523. ISSN 0006-4971 Boj, ...
Knockout Sudoku allows construction of whole-genome knockout collections for a wide range of microorganisms at a lower cost and increased speed, using combinatorial pooling, next-generation sequencing, and a Bayesian inference algorithm to process and annotate extremely large progenitor transposon insertion mutant collections. Knockout Sudoku is a method for the construction of whole-genome knockout collections for a wide range of microorganisms with as little as 3 weeks of dedicated labor and at a cost of ∼$10,000 for a collection for a single organism. The method uses manual 4D combinatorial pooling, next-generation sequencing, and a Bayesian inference algorithm to rapidly process and then accurately annotate the extremely large progenitor transposon insertion mutant collections needed to achieve saturating coverage of complex microbial genomes. This method is ∼100× faster and 30× lower in cost than the next comparable method (In-seq) for annotating transposon mutant collections by combinatorial
Gene trap vectors have been used in insertional mutagenesis in animal systems to clone genes with interesting patterns of expression. These vectors are designed to allow the expression of a reporter gene when the vector inserts into a transcribed region. In this paper we examine alternative splicing events that result in the expression of a GUS reporter gene carried on a Ds element which has been designed as a gene trap vector for plants. We have developed a rapid and reliable method based on PCR to study such events. Many splice donor sites were observed in the 3 Ac border. The relative frequency of utilisation of certain splice donor and acceptor sites differed between tobacco and Arabidopsis. A higher stringency of splicing was observed in Arabidopsis.. ...
Read Strategies for Mutagenesis and Gene Cloning Using Transposon Tagging and T-DNA Insertional Mutagenesis, Annual Review of Plant Biology on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.
The gene trap vector is introduced into ES cells by electroporation or retroviral infection and clones containing gene trap insertions are selected by antibiotic resistance. Antibiotic resistant clones are then picked into 96-well master plates, which are later replicated to generate frozen stocks for long-term storage of clones, cell lysates for RNA and DNA isolation followed by PCR-based gene identification techniques.. ...
Nusse R, van Ooyen A, Rijsewijk F, van Lohuizen M, Schuuring E, vant Veer L. Retroviral insertional mutagenesis in murine mammary cancer. Proc R Soc Lond B Biol Sci. 1985;226:3-13. Abstract ...
The IGTC focuses on creating resources of embryonic stem cells with gene trap insertions in every or most genes in the mouse genome. A brief description of the gene trap consortium is available online. The Gene Trap Consortium is also described in Nature Genetics 36: 543-544, 2004. Links to Databases of Gene Trapped ES Cell Lines:. ...
Systems and methods for isolating DNA molecules that can include the steps of: providing a transposon mutant collection, the transposon mutant collection being stored in a plurality of wells; dispatc
OFDM (orthogonal frequency division multiplexing) communication is very attractive for high data rate transmission especially in the frequency selective fa
A novel P[UAS] insertion line shows progressive behavioral defects.Crossing P[UAS]117 to the pdf-gal4 driver results in a significant decrease in the rhythmicit
Sallaud C, Gay C, Larmande P, Bès M, Piffanelli P, Piégu B, Droc G, Regad F, Bourgeois E, Meynard D et al.. 2004. High throughput T-DNA insertion mutagenesis in rice: a first step towards in silico reverse genetics. The Plant journal : for cell and molecular biology. 39(3):450-64. Abstract ...
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Insertion Verification by PCR:. We will make fly DNA from transgenic stocks and perform PCR to confirm insertion at the targeted site. $25 per line.. Virgin Collection:. ...
An insertion is an addition in which an atom in one reactant is inserted between two atoms bound to each other by a covalent bond in the other reactant.. ...
Transposon insertion site profiling chip (TIP-chip) was invented by Researchers at the Johns Hopkins High Throughput Biology Center. Tip-chip can be used to help identify otherwise elusive disease-causing mutations in the 97 percent of the genome long believed to be
Promoterless gene trap vectors have been widely used for high-efficiency gene targeting and random mutagenesis in embryonic stem (ES) cells. Unfortunately, such vectors are only effective for genes expressed in ES cells and this has prompted the development of expression-independent vectors. These polyadenylation (poly A) trap vectors employ a splice donor to capture an endogenous genes polyadenylation sequence and provide transcript stability. However, the spectrum of mutations generated by these vectors appears largely restricted to the last intron of target loci due to nonsense-mediated mRNA decay (NMD) making them unsuitable for gene targeting applications. Here, we present novel poly A trap vectors that overcome the effect of NMD and also employ RNA instability sequences to improve splicing efficiency. The set of random insertions generated with these vectors show a significantly reduced insertional bias and the vectors can be targeted directly to a 5′ intron. We also show that this ...
Looking for online definition of interrupted gene in the Medical Dictionary? interrupted gene explanation free. What is interrupted gene? Meaning of interrupted gene medical term. What does interrupted gene mean?
We are attempting to identify cellular oncogenes activated in mammary tumours by using the mouse mammary tumour virus (MMTV) as an insertional mutagen. MMTV, a retrovirus lacking a host cell-derived viral oncogene, induces adenocarcinomas of the mammary gland after a long latency period. The tumours are clonal outgrowths of cells carrying one or more integrated MMTV proviral copies. We have cloned an integrated MMTV provirus with its adjacent chromosomal DNA and we have established that the insertion site was part of a domain of the mouse genome in which MMTV proviruses are inserted in many different tumours. A gene within this domain, called int-1 is transcriptionally activated as a consequence of proviral integration. We have proposed that int-1 is a cellular oncogene for mammary tumours. Proviral activation of int-1 occurs in cis, over distances of up to 10 kilobases and is presumably caused by the transcriptional enhancer present on the MMTV long terminal repeat. The putative int-1 mammary ...
The PiggyBac (PB) transposon has emerged as a novel mutagenesis tool for understanding gene function and for phenotypic screening in eukaryotes
Carcinogenesis is a multistep process involving alterations in various cellular pathways. The critical genetic events driving the evolution of primary liver cancer, specifically hepatoblastoma and hepatocellular carcinoma (HCC), are still poorly understood. However, telomere stabilization is acknowledged as prerequisite for cancer progression in humans. In this project, human fetal hepatocytes were utilized as a cell culture model for untransformed, proliferating human liver cells, with...
Azotobacter vinelandii produces the biopolymer alginate, which has a wide range of industrial and pharmaceutical applications. A random transposon insertion mutant library was constructed from A. vinelandii ATCC12518Tc in order to identify genes and pathways affecting alginate biosynthesis, and abou.... ...
Multiple methods have been introduced over the past 30 years to identify the genomic insertion sites of transposable elements and other DNA elements that integrate into genomes. However, each of these methods suffer from limitations that can frustrate attempts to map multiple insertions in a single genome and to map insertions in genomes of high complexity that contain extensive repetitive DNA. I introduce a new method for transposon mapping that is simple to perform, can accurately map multiple insertions per genome, and generates long sequence reads that facilitate mapping to complex genomes. The method, called TagMap, for Tagmentation-based Mapping, relies on a modified Tn5 tagmentation protocol with a single tagmentation adaptor followed by PCR using primers specific to the tranposable element and the adaptor sequence. Several minor modifications to normal tagmentation reagents and protocols allow easy and rapid preparation of TagMap libraries. Short read sequencing starting from the adaptor ...
Transposon mutagenesis is one of the most widely used strategies to generate a large number of random mutations within a bacterial genome and then to precisely ...
Transposons have been used in invertebrates for transgenesis and insertional mutagens in genetic screens. We tested a functional transposon called Sleeping Beauty in the one-cell mouse embryo. In this report, we describe experiments in which transposon vectors were injected into one-cell mouse embryos with mRNA expressing the SB10 transposase enzyme. Molecular evidence of transposition was obtained by cloning of insertion sites from multiple transgenic mice produced by SB10 mRNA/transposon coinjection. We also demonstrate germ-line transmission and expression from transposed elements. This technique has promise as a germ-line transgenesis method in other vertebrate species and for insertional mutagenesis in the mouse ...
The non-coding regions of tumour cell genomes harbour a considerable fraction of total DNA sequence variation, but the functional contribution of these variants to tumorigenesis is ill-defined. Among these non-coding variants, somatic insertions are among the least well characterized due to challenges with interpreting short-read DNA sequences. Here, using a combination of Chip-seq to enrich enhancer DNA and a computational approach with multiple DNA alignment procedures, we identify enhancer-associated small insertion variants. Among the 102 tumour cell genomes we analyse, small insertions are frequently observed in enhancer DNA sequences near known oncogenes. Further study of one insertion, somatically acquired in primary leukaemia tumour genomes, reveals that it nucleates formation of an active enhancer that drives expression of the LMO2 oncogene. The approach described here to identify enhancer-associated small insertion variants provides a foundation for further study of these abnormalities ...
An alternative to insulin injections is the insulin pump. The pump is a computerized device, about the size of a beeper or pager, often worn on a belt or in a pocket. The pump delivers a continuous low (basal) dose of insulin through a cannula (a flexible plastic tube), which attaches to the body through a small needle inserted into the skin. The cannula is taped in place and the needle is removed. Common insertion sites on the body include the thighs, buttocks, abdomen, upper arms and other areas with fatty tissue.. When a person wearing a pump eats, she pushes a button on the pump to deliver an extra amount of insulin called a bolus to provide insulin for their food.. The advantages of the pump include:. ...
DNA signature tags (molecular barcodes) facilitate functional screens by identifying mutants in mixed populations that have a reduced or increased adaptation to a particular environment. Many innovative adaptations and refinements in the technology have been described since its original use with Salmonella; they have yielded a wealth of information on a broad range of biological processes--mainly in bacteria, but also in yeast and other fungi, viruses, parasites and, most recently, in mammalian cells. By combining whole-genome microarrays and comprehensive ordered libraries of mutants, high-throughput functional screens can now be achieved on a genomic scale.
Despite the recent completion of the human genome project, an ostensibly more difficult post-genomic challenge will be the functional annotation of all human genes and integration of this information into an operational cell-based model. Unfortunately, ...
An ampicillin resistance plasmid carrying the cloned repressor gene cII of the L phage (Salmonella lyphimurium) was conducted by Flac into an F- recipient. Two types of plaamids were isolated from Apr transconjugants. The majority of plasmids were dimers with one copy of Tn1000 inserted, the minority being monomers with one copy of Tn1000. This proportion remained unaltered when we used the Flac strain transformed with a monomeric form of the recombinant plasmid as a donor. An extensive oligomerization of pBR322-originating plasmids was proved in the presence of Flac; its presumable relationship to transposition-related processes is suggested.
Transposon insertion in ykyB increases the activation of an artificial ComK feedback loop.Strains PG401 (amyE::PcomG-lacZ-gfp, PcomG-comK, ΔmecA) and PG401-Tn4
An important aspect of functional genomics is understanding the phenotypic consequence of mutations in all genes in a genome. A comprehensive collection of gene knockouts allows a defined set of mutations to be systematically studied for more efficient association of genes to functions (reviewed in [1]). Multiple approaches have been used to develop comprehensive knockout resources. Biological differences between organisms make specific technologies such as homologous recombination, RNA interference, or insertional mutagenesis more useful in generating a resource for an individual species. In plants, a comprehensive knockout collection was generated for Arabidopsis thaliana via mutagenesis with insertion tags [2-5]. Genomic DNA flanking each tag was systematically amplified and sequenced from each mutant. These Flanking Sequence Tags (FSTs) index each mutant to the genome and are accessible through the SIGnAL T-DNA Express database, which links the mutant stocks to genome annotations [6]. A ...
Zebrafish is an excellent model animal to study vertebrate development by genetic approaches. Hundreds of mutations affecting various processes of development have been isolated by chemical mutagenesis and insertional mutagenesis using a pseudotyped retrovirus. However, useful transposon tools and m …
에탄올, 고온, 저해제 스트레스 상황에서 내성을 가지는 사카로마이세스 세레비지아에 균주는 에탄올 생산 산업을 위해 중요한 부분을 차지한다. 본 연구에서는 에탄올 15 % 까지 내성을 가지는 5개의 균주를 트랜스포존에 의해 돌연변이가 일어난 집단에서 테스트를 통해 분리했다. 5개중에 2개는 고온 (42 °C)에도 내성을 가지는 것을 조사되었고, 그 중에서 1개는 푸르푸랄 50 mM까지 내성을 가지는 것을 확인했다. 트랜스포존이 끼워 들어간 위치를 통해 7개의 추정 유전자 (CMP2, IMD4, SSK2, PPG1, DLD3, PAM1, MSN2)를 확인하고, 노던 블럿 분석법을 통해 CMP2와 IMD4, SSK2와 PPG1유전자가 동시에 하향조절 되었고, DLD3 유전자또한 하향조절, PAM1과 MSN2의 전사 해독 틀이 깨져있는 것을 확인 할 수 있었다. 이 7 유전자가 독립적으로 망가진 돌연변이는 에탄올 내성이 있었고, 그들 ...
These OVE#2489A mice harbor a mutation created by random insertion of the SB-sa-IRES-rtTA-pA-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2223). The donating investigator reports the phenotype of homozygous mice as: embryonic day (E)8 lethal.
These OVE#2524B mice harbor a mutation created by random insertion of the SB-cHS4core-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2229). The donating investigator reports the phenotype of homozygous mice as: renal agenesis.
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TY - JOUR. T1 - Insertional mutagenesis and cloning of the genes required for biosynthesis of the host-specific AK-toxin in the Japanese pear pathotype of Alternaria alternata. AU - Tanaka, Aiko. AU - Shiotani, Hiroshi. AU - Yamamoto, Mikihiro. AU - Tsuge, Takashi. N1 - Copyright: Copyright 2017 Elsevier B.V., All rights reserved.. PY - 1999/8. Y1 - 1999/8. N2 - The Japanese pear pathotype of Alternaria alternata causes black spot of Japanese pear by producing a host-specific toxin known as AK-toxin. Restriction enzyme-mediated integration (REMI) mutagenesis was used to tag genes required for toxin biosynthesis. Protoplasts of a wild-type strain were treated with a linearized plasmid along with the restriction enzyme used to linearize the plasmid. Of 984 REMI transformants recovered, three produced no detectable AK-toxin and lost pathogenicity on pear leaves. Genomic DNA flanking the integrated plasmid was recovered from one of the mutants. With the recovered DNA used as a probe, a cosmid clone ...
Transposons are mobile DNA segments that can disrupt gene function by inserting in or near genes. Here, we show that insertional mutagenesis by the PiggyBac transposon can be used for cancer gene discovery in mice. PiggyBac transposition in genetically engineered transposon-transposase mice induced cancers whose type (hematopoietic versus solid) and latency were dependent on the regulatory elements introduced into transposons. Analysis of 63 hematopoietic tumors revealed that PiggyBac is capable of genome-wide mutagenesis. The Piggybac screen uncovered many cancer genes not identified in previous retroviral or Sleeping Beauty transposon screens, including Spic, which encodes a PU.1-related transcription factor, and Hdac7, a histone deacetylase gene. PiggyBac and Sleeping Beauty have different integration preferences. To maximize the utility of the tool, we engineered 20 mouse lines to be compatible with both transposases in constitutive, tissue-, or temporal-specific mutagenesis. Mice with ...
We report an SB insertional mutagenesis screen to identify loci that cooperate with oncogenic Kras to drive pancreatic adenocarcinoma in the mouse. SB-driven pancreatic adenocarcinomas show all stages of mPanIN that develop into early noninvasive adenocarcinoma and finally, invasive, highly metastatic adenocarcinoma. Importantly, these tumors also have a high desmoplastic component, a predominant histopathic feature of human pancreatic cancer. The T2Onc3 transposon proved to be more potent than T2Onc2 in inducing metastatic tumors. The exact mechanism for the difference between the two transposons in driving pancreatic cancer is unknown. One possibility is that the location of the transposon concatemer donor influences the development of the tumor. The T2Onc3 donor resides on chromosome 9, and pancreatic cancer genes in close proximity may be frequently mutated in these tumors because of local hopping.. Both T2Onc2 and T2Onc3 cohorts exhibited considerably higher numbers of nonredundant ...
High-throughput analysis of genome-wide transposon mutant libraries is a powerful tool for (conditional) essential gene discovery. Recently, several next generation sequencing approaches, e.g. Tn-seq, INseq and TraDIS, have been developed that accurately map the site of transposon insertions by mutant-specific amplification and sequence readout of DNA flanking the transposon insertions site, assigning a measure of essentiality based on the number of reads per gene or per mutant. However, analysis of these large and complex datasets is hampered by the lack of an easy to use and automated tool for transposon insertion sequencing data ...
The extraintestinal pathogen, avian pathogenic E. coli (APEC), known to cause systemic infections in chickens, is responsible for large economic losses in the poultry industry worldwide. In order to identify genes involved in the early essential stages of pathogenesis, namely adhesion and colonization, Signature-tagged mutagenesis (STM) was applied to a previously established lung colonization model of infection by generating and screening a total of 1,800 mutants of an APEC strain IMT5155 (O2:K1:H5; Sequence type complex 95). The study led to the identification of new genes of interest, including two adhesins, one of which coded for a novel APEC fimbrial adhesin (Yqi) not described for its role in APEC pathogenesis to date. Its gene product has been temporarily designated ExPEC Adhesin I (EA/I) until the adhesin-specific receptor is identified. Deletion of the ExPEC adhesin I gene resulted in reduced colonization ability by APEC strain IMT5155 both in vitro and in vivo. Furthermore, complementation of
In this study, we created a fission yeast insertion mutant library in which all mutants were tagged with unique barcode sequences and stored as two readily available selection platforms. The 384-well mutant arrays allow genetic screens on individual mutants and can be extended to genetic approaches such as synthetic genetic array (SGA) [45, 46]. These mutant arrays have been used to identify mutants with four distinct phenotypes (Table 2) as well as strains that are hyper-sensitive to cancer chemotherapeutics camptothecin and bleomycin (Hale and Runge, unpublished data). In addition to 384-well mutant arrays, mutant pools of 1800 mutants are available for parallel analysis.. The insertion mutagenesis used in this study relied on random non-homologous recombination, where a vast majority of transformants have unstable, circularized vector DNA and only a small portion have stable insertions. To facilitate the collection of stable insertion mutants, we included low-dose 5-FOA in our initial ...
TY - JOUR. T1 - A Tn10 derivative (T-POP) for isolation of insertions with conditional (tetracycline-dependent) phenotypes. AU - Rappleye, Chad A.. AU - Roth, John R.. PY - 1997/9. Y1 - 1997/9. N2 - A new Tn10-based transposon has been constructed and used to isolate insertion mutations with tetracycline-conditional phenotypes. Classes of mutants include conditional lethal mutations, conditional auxotrophs, and conditional mutants of the eut (ethanolamine utilization) operon. The described mutations were made with a new derivative of Tn10dTet that we have called Tn10d(T-POP). Others have noted that transposon Tn10dTet directs weak tetracycline-inducible transcripts out of both ends of the element into adjacent sequences. We have increased this level of outward transcription from Tn10dTet by selecting deletion mutations within the element that presumably remove transcription-termination signals. Insertion of the Tn10d(T-POP) element within an operon disrupts the target gene and makes expression ...
In this communication, we report a novel strategy for the genetic manipulation of large viral DNA genomes. In a single step we cloned an infectious cytomegalovirus DNA as a bacterial artificial chromosome in E. coli and reconstituted virus progeny after transfection of the BAC plasmid into eukaryotic cells. This approach makes the CMV genome accessible to the genetic techniques established for E. coli. As an example for the power of the mutagenesis procedures, we performed a targeted insertion of four nucleotides into the 230-kb MCMV genome. In principle, any mutation (point mutations, insertions, and deletions) in any region of the genome can now be introduced using the described mutagenesis procedure. Moreover, other procedures, for example a random transposon mutagenesis of the CMV genome are conceivable. Multiple mutations can be introduced in consecutive rounds of mutagenesis without the need to reconstitute infectious viral intermediates. Construction of revertant genomes can be easily ...
Chapter 1: Introduction: Author: Anton Berns (Netherlands Cancer Institute) -- Chapter 2: Retroviral mutagenesis in mouse leukemia/lymphoma: Author: David Largaespada, Ph.D. (University of Minnesota) -- Chapter 3: MMTV models of breast cancer: Author: John Hilkens (NKI) -- Chapter 4: Retroviral mutagenesis in other organisms: Author: Michael Dvorak (Inst. Of Molecular Genetics, Prague, Czech Rep.) -- Chapter 5: Sleeping Beauty models of cancer: Author: Adam J. Dupuy, Ph.D. (University of Iowa) -- Chapter 6: Insertional mutagenesis in gene therapy patients: Author: David Williams, M.D. (Cincinnati Children?셲 Hospital) -- Chapter 7: Bioinformatics of high throughput insertional mutagenesis: Author: Keiko Akagi (NCI-Frederick ...
Active retrotransposons play important roles during evolution and continue to shape our genomes today, especially in genetic polymorphisms underlying a diverse set of diseases. However, studies of human retrotransposon insertion polymorphisms (RIPs) based on whole-genome deep sequencing at the population level have not been sufficiently undertaken, despite the obvious need for a thorough characterization of RIPs in the general population.|br| Herein, we present a novel and efficient computational tool named Specific Insertions Detector (SID) for the detection of non-reference RIPs. We demonstrate that SID is suitable for high depth whole-genome sequencing (WGS) data using paired-end reads obtained from simulated and real datasets. We construct a comprehensive RIP database using a large population of 90 Han Chinese individuals with a mean 68× depth per individual. In total, we identify 9342 recent RIPs, and 8433 of these RIPs are novel compared with dbRIP, including 5826 Alu, 2169 long interspersed
A fixed gene copy number is important for the in silico construction of engineered synthetic networks. However, the copy number of integrated genes depends on their genomic location. This gene dosage effect is rarely addressed in synthetic biology. Two studies in Escherichia colipresented conflicting data on the impact of gene dosage. Here, we investigate how genome location and gene orientation influences expression in Bacillus subtilis. An important difference with the E. coli studies is that we used an unbiased genome integration approach mediated by random transposon insertion. We found that there is a strong gene dosage effect in fast growing B. subtilis cells, which can amount to a 5-fold difference in gene expression. In contrast, gene orientation with respect to DNA replication direction does not influence gene expression. Our study shows that gene dosage should be taken into account when designing synthetic circuits in B. subtilis and presumably other bacteria.. ...
Institutions: Samuel Lunenfeld Research Institute We are performing gene trap-based expression and genotypic screens to generate new mouse mutations that will help delineate the molecular controls of specific developmental programs. Using a polyA trap vector with recombination sites for post-insertional manipulations, gene trap insertions are screened using multiplexed in vitro differentiation and induction assays. An expression profile is being generated at a rate of greater than 5000 per year. Sequence tags for all insertions demonstrating restricted expression patterns (about 20%) are now being generated. A database is being developed that will be searchable by expression pattern, sequence, and phenotype. The clones will be available as a resource to researchers worldwide. One of the clones identified in our ongoing screen, SNAG-1, is expressed by hematopoietic stem cells, neural endothelial cells, cardiomyocytes and sensory nerves. SNAG-1 mutant embryos die mid-gestation with neural vascular ...
The mini-Tn5 are mobile elements able to transpose to the chromosome from a delivery plasmid, by means of the transposase activity encoded by the tnp* gene, which is present in cis in the plasmid but external to the mini-Tn5 element. pUT mini-Tn5 vectors can be used to insert any DNA fragment in the bacterial chromosome, by cloning it within the mini-Tn5 region. Due to the loss of the tnp* gene after insertion, minitransposons are stably integrated into the chromosome and inherited. The different pUTmini-Tn5 vectors can then be used for repeated insertion events and therefore multiple insertions in the same strain are only limited by the availability of distinct selection markers.. Advantages:. ...
Bibliographic details on FLAGdb/FST: a database of mapped flanking insertion sites (FSTs) of Arabidopsis thaliana T-DNA transformants.
The Genetics Society of America (GSA), founded in 1931, is the professional membership organization for scientific researchers and educators in the field of genetics. Our members work to advance knowledge in the basic mechanisms of inheritance, from the molecular to the population level.. Online ISSN: 1943-2631. ...
Atarés Huerta, Alejandro; Moyano, Elena; Morales, Belén; Schleicher, Peter; García Abellán, José Osvaldo; Antón Martínez, María Teresa; García Sogo, Begoña; Pérez Martin, Fernando; Lozano, Rafael; Borja Flores, Francisco; Moreno Ferrero, Vicente; Bolarin Jimenez, Maria del Carmen; Pineda Chaza, Benito José (Springer Verlag (Germany), 2011) ...
A new paper in Cell Reports utilizes RNA-seq and Tn-seq (the tn in tn-seq stands for transposon) to map the transcriptional and fitness changes in bacterial gene networks in response to stressors, like nutrient depletion and antibiotics.. The transcriptional response measures changes in gene expression as measured by RNA-seq. The fitness or phenotypic response describes the importance of each gene to the response. This is measured by a different assay, Tn-seq, which takes advantage of transposon insertion to selectively inactivate genes in the bacterial genome. Those genes that are depleted in the stressor condition are determined to be high fitness (owing to the fact that the bacteria without those genes died under stress).. First, before even considering DE genes, they found that there is no correlation between a genes transcriptional abundance (not fold change) and its fitness. While most high-fitness genes were also high abundance, many more high-abundance genes were not high-fitness. ...
From the nomenclature guidelines: Newly generated Transposon insertions, especially those located in apparently intergenic regions, may also be given Ti (transposon insertion) names. These consist of the designation identifying the laboratory of origin, the two letters Ti, and a number, all italicized. Example: eTi13 is an insertion of a Mos transposon into an intergenic region on LGIII ...
A DNA transposon, or jumping gene, controls its amplification within a genome through a competition between the enzyme multimers that are responsible for its mobility.
Interestingly there is no rule such as the higher evolved an organism the more transposable elements. Although we find more transposable elements in higher evolved organisms this rule can not be maintained if compared single species as frogs and humans.. Transposons most often code for transposase the enzyme responsible for the transposon dislocation. As Transposons are so common in a genome it is not surprising that transposases are the most common genes in a genome [2].. ...
Hard hitting drums and a huge bass lead that will have you up in arms and ready to defend New York from the Cleaners and LMB! Equip those new mods and party up! WERE HEADED TO THE DARK ZONE!. ...
The Rosa26 gene locus is well suited for gene over-expression. Reduced development time and cost with ready-to-use targeting vector.
Dr. performed reduction and percutaneous rush pins insertion on open fracture of humeral neck. What is a rush pin considered? What code wo
A summary of The Insertion Sort Algorithm in s Insertion Sort. Learn exactly what happened in this chapter, scene, or section of Insertion Sort and what it means. Perfect for acing essays, tests, and quizzes, as well as for writing lesson plans.
It is possible that there is an internal rearrangement of the MiMIC transposon involving the 3prime end. May be segregating SM6a ...
Mutations can also be generated by insertional mutagenesis. For example, transposable elements containing a marker are ... or insertional mutagenesis (e.g. transposable elements). Subsequent breeding takes place, mutant individuals are isolated, and ... This type of saturation mutagenesis within classical experiments was used to define sets of genes that were a bare minimum for ...
For: Carlson, Corey M.; Largaespada, David A. (2005). "Insertional mutagenesis in mice: New perspectives and tools". Nature ...
Carlson CM, Largaespada DA (July 2005). "Insertional mutagenesis in mice: new perspectives and tools". Nature Reviews Genetics ...
2003). "Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana". Science. 301 (5633): 653-657. doi:10.1126/science.1086391 ... but also due to many new mutagenesis programs encouraged by the development of Arabidopsis as a major model organism ...
This is advantageous as insertional mutagenesis is avoided. It is disadvantageous in that the progeny of the cell will lose the ... With this approach there is increased risk of insertional mutagenesis; however, the risk can be reduced by using an integrase- ...
This process is referred to as insertional mutagenesis or transposon mutagenesis. When a gene is inactivated by insertion of a ... Carlson CM, Largaespada DA (July 2005). "Insertional mutagenesis in mice: new perspectives and tools". Nat. Rev. Genet. 6 (7): ...
Methylation makes the resistant gene inactive; this is called insertional inactivation or insertional mutagenesis. For example ...
In 1994, a new allele of reeler was obtained by means of insertional mutagenesis. This provided the first molecular marker of ... "Isolation of an allele of reeler by insertional mutagenesis". Proceedings of the National Academy of Sciences of the United ...
... and insertional mutagenesis. Among these methods, insertional mutagenesis was proved to be very good and successful approach. ... At first, T-DNA was applied for insertional mutagenesis. However, using transposable element can bring more advantages. ...
discovered the sleepless gene in fruit flies through insertional mutagenesis. Mutations in the sleepless gene caused the flies ...
This technique is used in transgenesis and insertional mutagenesis research fields. The Sleeping Beauty transposon system is an ...
causing massive [[transposon]] insertional mutagenesis that could explain the phenotypic diversification. Understanding ... "Evaluating risks of insertional mutagenesis by DNA transposons in gene therapy". Translational Research. pp. 265-283. doi: ...
Stewart M, Mackay N, Hanlon L, Blyth K, Scobie L, Cameron E, Neil JC (Jun 2007). "Insertional mutagenesis reveals progression ... Studies using high throughput viral insertional mutagenesis analysis also revealed that JDP2 functions as an oncogene. JDP2- ... "Tumor model-specific proviral insertional mutagenesis of the Fos/Jdp2/Batf locus". Virology. 337 (2): 353-64. doi:10.1016/j. ... "Identification of oncogenes collaborating with p27Kip1 loss by insertional mutagenesis and high-throughput insertion site ...
Rabinowitz JE, Xiao W, Samulski RJ (December 1999). "Insertional mutagenesis of AAV2 capsid and the production of recombinant ... Muralidhar S, Becerra SP, Rose JA (January 1994). "Site-directed mutagenesis of adeno-associated virus type 2 structural ... which present the threat of a random insertion and of mutagenesis, which is sometimes followed by development of a cancer. The ...
Used in insertional mutagenesis *Insertional mutagenesis uses the features of a TE to insert a sequence. In most cases this is ... Insertional mutagenesis uses the features of a TE to insert a sequence. In most cases this is used to remove a DNA sequence or ... Mechanisms of mutagenesis[edit]. TEs are mutagens and their movements are often the causes of genetic disease. They can damage ... The Sleeping Beauty transposon system has been used extensively as an insertional tag for identifying cancer genes.[52] ...
Used in insertional mutagenesis Insertional mutagenesis uses the features of a TE to insert a sequence. In most cases this is ... Insertional mutagenesis uses the features of a TE to insert a sequence. In most cases this is used to remove a DNA sequence or ... The Sleeping Beauty transposon system has been used extensively as an insertional tag for identifying cancer genes. The Tc1/ ... Used for analysis of gene expression and protein functioning in signature-tagging mutagenesis. This analytical tool allows ...
T-DNA tagging mutagenesis involves screening of populations by T-DNA insertional mutations. Collections of known T-DNA ... The same procedure of T-DNA transfer can be used to disrupt genes via insertional mutagenesis. Not only does the inserted T-DNA ... Krysan PJ, Young JC, Sussman MR (December 1999). "T-DNA as an insertional mutagen in Arabidopsis". The Plant Cell. 11 (12): ... Feldmann KA (1991-07-01). "T-DNA insertion mutagenesis in Arabidopsis: mutational spectrum". The Plant Journal. 1 (1): 71-82. ...
Because of that property, transposons have been manipulated for use in insertional mutagenesis. The development of microbial ... Transposon insertion sequencing (Tn-seq) combines transposon insertional mutagenesis with massively parallel sequencing (MPS) ... Tn-seq is useful for both the study of a single gene's fitness as well as gene interactions Signature-tagged mutagenesis (STM) ... Kleckner N, Chan RK, Tye BK, Botstein D (October 1975). "Mutagenesis by insertion of a drug-resistance element carrying an ...
In mice, a dysfunctional MAOA gene is created through insertional mutagenesis (called 'Tg8'). Tg8 is a transgenic mouse strain ...
Youngman, PJ; Perkins, JB; Losick, R (1983). "Genetic transposition and insertional mutagenesis in Bacillus subtilis with ... Site-directed mutagenesis is a technique that has been around since the 1970s. The early days of research in this field yielded ... Site directed mutagenesis is a valuable technique that allows for the replacement of a single base in an oligonucleotide or ... Site-directed mutagenesis can be useful for many different reasons. A single base pair replacement, could change a codon, and ...
Candidates are designed to have improved folding and translation efficiency via insertional mutagenesis. The Moderna COVID-19 ...
"Insertional mutagenesis identifies drivers of a novel oncogenic pathway in invasive lobular breast carcinoma". Nature Genetics ... Miyamoto CA, Fischman DA, Reinach FC (October 1999). "The interface between MyBP-C and myosin: site-directed mutagenesis of the ... a high-throughput mutagenesis project to generate and distribute animal models of disease to interested scientists. Male and ...
They discovered that the virus activated cellular oncogenes by several mechanisms of insertional mutagenesis. When Jim ...
The SB transposon is a powerful tool for insertional mutagenesis in many vertebrate species. It recently exhibited especial ... The first published attempt had a success rate of less than 1%. Alternatively, ENU mutagenesis is a common random mutagenesis ... in vivo SB insertional mutagenesis allows multiple mutations to be quickly and easily generated in a single animal, and in a ... ENU mutagenesis involves using a chemical, N-ethyl-N-nitrosourea (ENU), to create single base changes in the genome. ENU ...
"Systems Biology-Based Investigation of Cellular Antiviral Drug Targets Identified by Gene-Trap Insertional Mutagenesis". PLOS ...
Fli-1 is activated through retroviral insertional mutagenesis in 90% of F-MuLV-induced erythroleukemias. The constitutive ... insertional activation of a new member of the ets gene family, Fli-1, closely linked to c-ets-1". Genes & Development. 5 (6): ...
Insertional mutagenesis is when transposons function as vectors to help remove and integrate genetic sequences. Given their ... Forward genetics methods) There are two main ways to utilise these tools: Fly Transformation, and Insertional Mutagenesis, each ... transposons have been used successfully in plants and invertebrate subjects through insertional mutagenesis and insertional ... Currently transposons can be used in genetic research and recombinant genetic engineering for insertional mutagenesis. ...
Levy LS, Lobelle-Rich PA, Overbaugh J (1993). "flvi-2, a target of retroviral insertional mutagenesis in feline thymic ...
Carlson C.M.; Largaespada D.A. (2005). "Insertional mutagenesis in mice: new perspectives and tools". Nature Reviews Genetics. ... in a process called insertional mutagenesis; transposase-mediated excision of the DNA transposon restores gene function. This ... Researchers use them as a means of mutagenesis. In this context, a TE jumps into a gene and produces a mutation. The presence ... The Sleeping Beauty transposon system has been used extensively as an insertional tag for identifying cancer genes.[46] ...
"Identification of oncogenes collaborating with p27Kip1 loss by insertional mutagenesis and high-throughput insertion site ...
This gives rise to a concern that insertional mutagenesis due to integration into the host genome might lead to cancer or ...
... one of the structural genes of the NiFe uptake hydrogenase was inactivated by insertional mutagenesis, and the mutant strain ...
Insertional mutagenesis. *Loss-of-Function Mutations. *Gain-of-Function Mutations. References[edit]. *^ Banavali, Nilesh K. ( ...
Through these collections, insertional mutants are available for most genes in A. thaliana. ... thaliana mutants that they generated using X-ray mutagenesis. Laibach continued his important contributions to A. thaliana ...
"Transposon-mediated insertional mutagenesis revealed the functions of animal cellulose synthase in the ascidian Ciona ...
Additionally, because EFV has two promoters, insertional mutagenesis would be safe and more accurate than other retroviruses ...
... establishing resources for genetic mapping and insertional mutagenesis, and providing physical maps that underpinned the ...
Labarre J, Chauvat F, Thuriaux P (June 1989). "Insertional mutagenesis by random cloning of antibiotic resistance genes into ...
... it completely eliminates the risk of genomic integration and insertional mutagenesis inherent to all DNA-based methodologies." ...
... a gene activated by insertional mutagenesis in mouse models of leukemia. Blood 103: 3897-3904, 2004. Leukemia/Bone Marrow ...
Other advantages are human to human transmission has never been reported, it has a safer spectrum of insertional mutagenesis ...
... and it also avoids possible insertional mutagenesis caused by integration into host chromosomes by viral vector. Animal and ...
PCR mutagenesis Insertional mutagenesis Signature tagged mutagenesis Transposon mutagenesis Sequence saturation mutagenesis ... Adaptive mutagenesis has been defined as mutagenesis mechanisms that enable an organism to adapt to an environmental stress. ... Early methods of mutagenesis produced entirely random mutations; however, modern methods of mutagenesis are capable of ... Modern laboratory techniques used to generate these mutations include: Directed mutagenesis Site-directed mutagenesis/ ...
... for T-DNA insertional mutagenesis". Plant Biotechnology Journal. 6 (5): 236-45. doi:10.1111/j.1467-7652.2007.00308.x. PMID ... Thole, Vera; Peraldi, Antoine; Worland, Barbara; Nicholson, Paul; Doonan, John H.; Vain, Philippe (2012). "T-DNA mutagenesis in ... Engvild, Kjeld C. (March 2005). "Mutagenesis of the Model Grass Brachypodium distachyon with Sodium Azide". Risø National ... in Brachypodium distachyon T-DNA insertional mutants". Nature Protocols. 4 (5): 650-61. doi:10.1038/nprot.2009.32. PMID ...
"Induction of antiestrogen resistance in human breast cancer cells by random insertional mutagenesis using defective ...
Also, it does not integrate into the genome and therefore does not have the risk of insertional mutagenesis. Moreover, IVT mRNA ... mRNAs do not integrate into the genome and therefore do not have the risk of insertional mutagenesis, making them suitable for ...
AAV also has little risk for insertional mutagenesis, a common problem when dealing with viral vectors, as its transgenes are ...
insertional mutagenesis synonyms, insertional mutagenesis pronunciation, insertional mutagenesis translation, English ... dictionary definition of insertional mutagenesis. Noun 1. insertional mutagenesis - a mutation caused by the insertion of ... Insertional mutagenesis - definition of insertional mutagenesis by The Free Dictionary ... a href=,insertional mutagenesis,/a,. *Facebook ...
Gene Discovery by MMTV Mediated Insertional Mutagenesis. In: Dupuy A., Largaespada D. (eds) Insertional Mutagenesis Strategies ... Retroviral insertional mutagenesis (IM) screens provide one of the most efficient tools to identify genes involved in ... 2007). MMTV insertional mutagenesis identifies genes, gene families and pathways involved in mammary cancer. Nature of Genetics ... Lee, F. S., Lane, T. F., Kuo, A., Shackleford, G. M., & Leder, P. (1995). Insertional mutagenesis identifies a member of the ...
Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana. By José M. Alonso, Anna N. Stepanova, Thomas J. Leisse, ... Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana. By José M. Alonso, Anna N. Stepanova, Thomas J. Leisse, ... Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana Message Subject. (Your Name) has forwarded a page to you from ...
... Testing Status of Insertional Mutagenesis - Definitive Vector Study ... Insertional mutagenesis studies using various vector exposures (LTR; uORF LTR, SIN, EFS LTR, and P(-) SIN vectors. ...
... have been altered by linker insertional mutagenesis in order to define critical components of virion assembly and infectivity. ... Insertional mutagenesis of AAV2 capsid and the production of recombinant virus Virology. 1999 Dec 20;265(2):274-85. doi: ... The structural genes of adeno-associated virus serotype 2 (AAV2) have been altered by linker insertional mutagenesis in order ...
Cloning by insertional mutagenesis of a cDNA encoding Caenorhabditis elegans kinesin heavy chain. N Patel, D Thierry-Mieg, and ... Cloning by insertional mutagenesis of a cDNA encoding Caenorhabditis elegans kinesin heavy chain ... Cloning by insertional mutagenesis of a cDNA encoding Caenorhabditis elegans kinesin heavy chain ... Cloning by insertional mutagenesis of a cDNA encoding Caenorhabditis elegans kinesin heavy chain ...
Insertional Mutagenesis of Genes Required for Seed Development in Arabidopsis thaliana. John McElver, Iris Tzafrir, George Aux ... Insertional Mutagenesis of Genes Required for Seed Development in Arabidopsis thaliana. John McElver, Iris Tzafrir, George Aux ... Insertional Mutagenesis of Genes Required for Seed Development in Arabidopsis thaliana. John McElver, Iris Tzafrir, George Aux ... 1996 Insertional mutagenesis to dissect embryonic development in Arabidopsis, pp. 51-76 in Embryogenesis: The Generation of a ...
Protecting the yeast genome from Ty1 insertional mutagenesis:. The analyses of Ty1 insertions at SUF16 and CAN1 in different ... Chromatin-Associated Genes Protect the Yeast Genome From Ty1 Insertional Mutagenesis Message Subject (Your Name) has forwarded ... Chromatin-Associated Genes Protect the Yeast Genome From Ty1 Insertional Mutagenesis. Katherine M. Nyswaner, Mary Ann Checkley ... Chromatin-Associated Genes Protect the Yeast Genome From Ty1 Insertional Mutagenesis. Katherine M. Nyswaner, Mary Ann Checkley ...
Validation-based insertional mutagenesis identifies lysine demethylase FBXL11 as a negative regulator of NFκB. Tao Lu, Mark W. ... While MLV-based insertional mutagenesis has a long history in gene discovery, we understand HIV-based insertion less well. ... Validation-based insertional mutagenesis identifies lysine demethylase FBXL11 as a negative regulator of NFκB ... Validation-based insertional mutagenesis identifies lysine demethylase FBXL11 as a negative regulator of NFκB ...
Gene therapy medicinal products, insertional mutagenesis, integration. Description. This document discusses the factors ... the strategies to reduce the risk associated to insertional mutagenesis and the assays to evaluate vector oncogenesis at the ...
ATMT-based approaches such as random insertional mutagenesis and targeted knockout are widely used for gene functional analysis ... Here, we describe a protocol for the identification of pathogenicity and virulence genes through random insertional mutagenesis ...
Efficient insertional mutagenesis in lactococci and other gram-positive bacteria.. E Maguin, H Prévost, S D Ehrlich, A Gruss ... Efficient insertional mutagenesis in lactococci and other gram-positive bacteria. Message Subject (Your Name) has forwarded a ... High-frequency random insertion (of about 1%) obtained with this system in Lactococcus lactis allows efficient mutagenesis, ... the study of chromosomal genes and their regulation is limited by the lack of an efficient transposon mutagenesis system. We ...
The mouse cystic fibrosis transmembrane conductance regulator gene was disrupted in embryonal stem cells using an insertional ... Cystic Fibrosis in the Mouse by Targeted Insertional Mutagenesis Nature. 1992 Sep 17;359(6392):211-5. doi: 10.1038/359211a0. ... This insertional mouse mutation provides a valid model system for the development and testing of therapies for cystic fibrosis ... The mouse cystic fibrosis transmembrane conductance regulator gene was disrupted in embryonal stem cells using an insertional ...
Challenges and Opportunities for Horticulture in the World INSERTIONAL MUTAGENESIS IN THE DIPLOID STRAWBERRY (FRAGARIA VESCA) ... INSERTIONAL MUTAGENESIS IN THE DIPLOID STRAWBERRY (FRAGARIA VESCA). Authors: R.E. Veilleux, T. Oosumi, P.A. Wadl, A.J. Baxter, ... Insertional mutant collections are essential tools in plant genomic research and functional genomics. Extensive collections in ... We have therefore undertaken a program to develop an insertional mutant collection of the diploid strawberry, Fragaria vesca, ...
Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of ... that can be used for in vitro insertional mutagenesis. Each of these DNA fragments carries a different antibiotic or Hg2+ ... Thus, the selection and/or characterization of omega insertional mutations can be carried out in these bacterial species. ...
... mutagenesis was used to identify mutants of Magnaporthe grisea impaired in pathogenicity. Three REMI protocols were evaluated ... Restriction enzyme-mediated DNA integration (REMI) mutagenesis was used to identify mutants of Magnaporthe grisea impaired in ...
Insertional Mutagenesis Risk Following Aav Vector-mediated Gene Transfer Author: Hojun Li, University of Pennsylvania School of ... Insertional Mutagenesis Risk Following Aav Vector-mediated Gene Transfer. Winning Abstracts from the 2010 Medical Student ... Insertional Mutagenesis Risk Following Aav Vector-mediated Gene Transfer. Author: Hojun Li, University of Pennsylvania School ... Insertional Mutagenesis Risk Following Aav Vector-mediated Gene Transfer. Winning Abstracts from the 2010 Medical Student ...
Mouse genetic corneal disease resulting from transgenic insertional mutagenesis Message subject: (Your Name) has forwarded a ...
Another possibility is that the transgenic insertional mutagenesis affected a regulatory gene, such as Pax6 or a similar gene. ... Rijkers T, Peetz A, Ruther U. Insertional mutagenesis in transgenic mice. Transgenic Res 1994;3:203-15. ... Conclusion: Eye abnormality in T27aT15 mice results from random insertional mutagenesis of the transgene as it was only ... Insertional mutation of classical and novel genes in transgenic mice. Trends Genet 1992;8:341-4. ...
Lymphomagenesis in SCID-X1 mice following lentivirus-mediated phenotype correction independent of insertional mutagenesis and ... Lymphomagenesis in SCID-X1 mice following lentivirus-mediated phenotype correction independent of insertional mutagenesis and ... Extensive analyses failed to implicate insertional mutagenesis or gammac overexpression as the underlying mechanism. These ... "Lymphomagenesis in SCID-X1 Mice Following Lentivirus-mediated Phenotype Correction Independent of Insertional Mutagenesis and ...
Genetic analysis of the pAD1 hemolysin/bacteriocin determinant in Enterococcus faecalis: Tn917 insertional mutagenesis and ... Genetic analysis of the pAD1 hemolysin/bacteriocin determinant in Enterococcus faecalis: Tn917 insertional mutagenesis and ... Genetic analysis of the pAD1 hemolysin/bacteriocin determinant in Enterococcus faecalis: Tn917 insertional mutagenesis and ... Genetic analysis of the pAD1 hemolysin/bacteriocin determinant in Enterococcus faecalis: Tn917 insertional mutagenesis and ...
... Wu J, Keng VW, Patmore DM ... Home , Research and Education , Find a Scientific Publication , Insertional Mutagenesis Identifies A Stat3arid1b Catenin ... we used unbiased insertional mutagenesis screening, mouse models, and molecular analyses. We mapped an Nf1-Stat3-Arid1b/β- ... we used unbiased insertional mutagenesis screening, mouse ...
Sleeping Beauty insertional mutagenesis to characterize the process of metastasis. Sleeping Beauty insertional mutagenesis may ... Limits and Advantages of Insertional Mutagenesis Systems. Comparing insertional mutagenesis with other cancer gene discovery ... Different mechanisms of gene deregulation by insertional mutagenesis. Vector integration can induce insertional mutagenesis by ... thus providing a kind of spontaneous insertional mutagenesis screening. Insertional mutagenesis has also been shown to occur in ...
A Dinculescu, JH McDowell, S Amici, D Dugger, N Richards, PA Hargrave, WC Smith; An Insertional Mutagenesis and Immunochemical ... An Insertional Mutagenesis and Immunochemical Analysis of Visual Arrestin Interaction with Rhodopsin ... An Insertional Mutagenesis and Immunochemical Analysis of Visual Arrestin Interaction with Rhodopsin ... Our goal was to further probe the structural determinants of arrestin function by using a combined insertional mutagenesis and ...
1 Vpr in a central helical domain and probed regions of Vpx by insertional mutagenesis. A predicted loop between helices two ... Other insertional mutants in the predicted loop and in a linker region between the central domain and the C-terminus may be ... 1 Vpr in a central helical domain and probed regions of Vpx by insertional mutagenesis. A predicted loop between helices two ... Other insertional mutants in the predicted loop and in a linker region between the central domain and the C-terminus may be ...
"Molecular mechanisms of insertional mutagenesis in yeasts and mycelium fungi, Russian Journal of Genetics" on DeepDyve, the ... Molecular mechanisms of insertional mutagenesis in yeasts and mycelium fungi. Molecular mechanisms of insertional mutagenesis ... Molecular mechanisms of insertional mutagenesis in yeasts and mycelium fungi. Dmytruk, K.; Sibirny, A. ... "Molecular mechanisms of insertional mutagenesis in yeasts and mycelium fungi." Russian Journal of Genetics 43.8 (2007): 835-845 ...
1995) Retroviral insertional mutagenesis of a target allele in a heterozygous murine cell line. Proc. Natl. Acad. Sci. USA 82: ... 1994) Generation of Chinese hamster ovary cell glycosylation mutants by retroviral insertional mutagenesis. J. Biol. Chem. 269: ... Generation of SV-resistant CHO mutant cells by retroviral insertional mutagenesis.CHO-22 cells were infected with U3Neo Mo-MuLV ... We have shown that cells resistant to SV infection can be developed by retroviral insertional mutagenesis. The mutant cells ...
Directed mutagenesis Insertion (genetics) PCR mutagenesis Site-directed mutagenesis Transposon mutagenesis Biffi A, et al., ... insertional mutagenesis is the creation of mutations of DNA by addition of one or more base pairs. Such insertional mutations ... Insertional mutagenesis is possible whether the virus is of the self-inactivating types commonly used in gene therapy or ... Insertional+mutagenesis at the US National Library of Medicine Medical Subject Headings (MeSH) Diagram at gene-technology- ...
Low dose ionizing radiation strongly stimulates insertional mutagenesis in a γH2AX dependent manner English version ... 1. Uren AG, Kool J, Berns A, van Lohuizen M. Retroviral insertional mutagenesis: past, present and future. Oncogene. Nature ... Vyšlo v časopise: Low dose ionizing radiation strongly stimulates insertional mutagenesis in a γH2AX dependent manner. PLoS ... Low dose ionizing radiation strongly stimulates insertional mutagenesis in a γH2AX dependent manner ...
Female gametophytic mutants of Arabidopsis thaliana identified in a gene trap insertional mutagenesis screen ... Download PDF Female gametophytic mutants of Arabidopsis thaliana identified in a gene trap insertional mutagenesis screen. ... Female gametophytic mutants of Arabidopsis thaliana identified in a gene trap insertional mutagenesis screen. International ...
  • We describe a highly efficient use of lentiviral validation-based insertional mutagenesis (VBIM) to generate large populations of mammalian cells in which a strong promoter is inserted into many different genomic loci, causing greatly increased expression of downstream sequences. (
  • To improve the features of the previous reversible promoter insertional technique, we designed a set of lentiviral validation-based insertional mutagenesis (VBIM) vectors that extend applications to nearly any mammalian cell, even cells that are not dividing. (
  • Our laboratory has developed a novel method, called VBIM, for validation-based insertional mutagenesis, for inserting strong cytomegalovirus (CMV) promoters approximately randomly into the genomes of mammalian cells in tissue culture. (
  • In forward genetics, mutations can be induced by using chemical mutagens, insertional mutagens or through the delivery of diverse libraries containing, for example, cDNAs, shRNAs, or genetic suppressor elements, with each approach creating distinct genetic changes ( 1 - 4 ). (
  • Insertional mutagenesis has been used as a functional forward genetics screen for the identification of novel genes involved in the pathogenesis of human cancers. (
  • When insertional mutagenesis induces the deregulation of oncogenes or tumor suppressor genes (TSG), it can cause cell transformation, and indeed insertional mutagenesis has been widely exploited for forward genetics screenings aimed at identifying novel cancer genes. (
  • download derive the download insertional mutagenesis strategies in cancer genetics kind brucellosis. (
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  • Emerging innovative genetic strategies, such as somatic genetics, modifier screens and humanized mice, in combination with whole-genome mutagenesis will dramatically broaden the utility of the mouse. (
  • What are the three broad steps of forward genetics (phenotype driven mutagenesis)? (
  • The 350 tagged embryo-defective ( emb ) mutants identified to date represent a significant advance toward saturation mutagenesis of EMB genes in Arabidopsis. (
  • The present study describes a system of inducible insertional mutagenesis based on the Ac-Ds family of transposons for targeted tagging in Arabidopsis thaliana. (
  • These results establish the feasibility of our approach for localized saturation mutagenesis in Arabidopsis. (
  • Fedoroff, Nina V. / Resources for targeted insertional and deletional mutagenesis in Arabidopsis . (
  • We describe a new resource for targeted insertional mutagenesis in Arabidopsis using a maize (Zea mays) Activator/Dissociation (Ds) two-element system. (
  • We are currently evaluating several hundred insertional mutants bearing the GFP reporter gene and hygromycin resistance for antibiotic selection. (
  • Single gene insertional mutants in the T1 generation have expressed unusual phenotypes for leaf morphology, anthocyanin development, flower morphology and plant habit. (
  • Restriction enzyme-mediated DNA integration (REMI) mutagenesis was used to identify mutants of Magnaporthe grisea impaired in pathogenicity. (
  • Other insertional mutants in the predicted loop and in a linker region between the central domain and the C-terminus may be useful as sites of intramolecular tags as they could be packaged adequately and retained preintegration complex associated integration activity in a serum starvation assay. (
  • Nuclear transformation to complement an Arg- phenotype was used to generate insertional mutants, and Cah1 expression and having high CO2-requiring (HCR) phenotype were used as reporters for induction of genes involved in acclimation to limiting CO 2. (
  • The combined approaches of embryonic stem cell-based technologies, chemical and insertional mutagenesis have enabled the systematic interrogation of the mouse genome with the aim of creating, for the first time, a library of mutants in which every gene is disrupted. (
  • This DNA is then transferred into P. aeruginosa and forced into the chromosome via homologous recombination, producing in a single step a random set of insertional mutants along a desired region of the chromosome. (
  • In this study, we developed a simple method for inducing robust monokaryotic fruiting and combined the assay with Agrobacterium tumefaciens insertional mutagenesis to screen for hyphal mutants. (
  • However, insertional mutants using these methods have not been reported. (
  • We report here development of an enhancer trap method by using the Tol2 transposable element and identification and characterization of insertional mutants. (
  • however, they are not flexible with respect to the kinds of mutants generated, nor are they as specific as later methods of site-directed mutagenesis and therefore have some degree of randomness. (
  • In molecular biology, insertional mutagenesis is the creation of mutations of DNA by addition of one or more base pairs. (
  • Such insertional mutations can occur naturally, mediated by viruses or transposons, or can be artificially created for research purposes in the lab. (
  • Thus, the selection and/or characterization of omega insertional mutations can be carried out in these bacterial species. (
  • To begin to identify cellular factors that influence the outcome of SV infection we have used a retroviral gene trap strategy that incorporates the use of a selectable marker to enrich for cells with insertional mutations ( 6 , 42 ) to generate mutant Chinese hamster ovary (CHO) cells resistant to SV. (
  • Application of this method to extend the analysis of the alginate regulatory region, using a cloned 6.2-kb fragment with the algr gene and the previously uncharacterized flanking regions, produced several insertional mutations. (
  • Mohr, CD & Deretic, V 1990, ' Gene-scrambling mutagenesis: Generation and analysis of insertional mutations in the alginate regulatory region of Pseudomonas aeruginosa ', Journal of bacteriology , vol. 172, no. 11, pp. 6252-6260. (
  • We thus demonstrated that the transposon-mediated enhancer trap approach can indeed create insertional mutations in developmental genes. (
  • In molecular biology, mutagenesis is an important laboratory technique whereby DNA mutations are deliberately engineered to produce libraries of mutant genes, proteins, strains of bacteria, or other genetically modified organisms. (
  • Early approaches to mutagenesis relied on methods which produced entirely random mutations. (
  • A variation of this method for integrating non-biased mutations in a gene is sequence saturation mutagenesis. (
  • Prior to the development site-directed mutagenesis techniques, all mutations made were random, and scientists had to use selection for the desired phenotype to find the desired mutation. (
  • Therefore, CHO cells appear to be a cell line appropriate for genetic mutant isolation by using retroviruses as insertional mutagens. (
  • Retroviruses and insertional mutagenesis in mice: proviral integratio" by P Soriano, T Gridley et al. (
  • Retroviruses and insertional mutagenesis in mice: proviral integration at the Mov 34 locus leads to early embryonic death. (
  • These retroviruses are divided into acute (retroviruses directly transducing the mutated form of a host proto-oncogene) and nonacute (not containing a virally transduced oncogene) that induce oncogenic transformation through the insertional mutagenesis. (
  • This is known as insertional mutagenesis. (
  • This adverse event, known as insertional mutagenesis, has become a major hurdle in the field. (
  • Point mutation was found in the vast majority of all patients, insertional mutagenesis in 4 cases. (
  • This insertional mouse mutation provides a valid model system for the development and testing of therapies for cystic fibrosis patients. (
  • Extrachromosomal DNA can integrate into the genome with no sequence specificity producing an insertional mutation. (
  • Mutagenesis where the mutation is caused by the introduction of foreign DNA sequences into a gene or extragenic sequence. (
  • A previously existing insertional mutation ( Gtl2lacZ ), and a targeted deletion in which the Gtl2 upstream region was replaced by a Neo cassette ( Gtl2Δ5'Neo ), display partial lethality and dwarfism upon paternal inheritance. (
  • Here we demonstrate the feasibility of conducting site-specific mutagenesis for disrupting virulence genes in C. caviae, an agent of guinea pig inclusion conjunctivitis that was recently identified as a zoonotic agent in cases of severe community-acquired pneumonia. (
  • Not all integrating viruses cause insertional mutagenesis, however. (
  • The objective of this work is to develop an in vitro assay to measure the frequency with which retroviral and lentiviral vectors cause insertional mutagenesis and to analyse the mechanisms by which this occurs. (
  • Insertional Mutagenesis Identifies a STAT3/Arid1b/β-catenin Pathway Driving Neurofibroma Initiation. (
  • Insertional mutagenesis identifies genes that promote the immortalization of primary bone marrow progenitor cells. (
  • Retroviral insertional mutagenesis in telomerase-immortalized hepatocytes identifies RIPK4 as novel tumor suppressor in human hepatocarcinogenesis. (
  • Genetic analysis of the pAD1 hemolysin/bacteriocin determinant in Enterococcus faecalis: Tn917 insertional mutagenesis and cloning. (
  • To elucidate critical downstream genetic events driving further transformation of immortalized liver cells, we used retroviral insertional mutagenesis as an unbiased approach to induce genetic alterations. (
  • utilized a tumor-prone mouse model to identify the genetic determinants of insertional mutagenesis (see the related article beginning on page 964). (
  • High-frequency random insertion (of about 1%) obtained with this system in Lactococcus lactis allows efficient mutagenesis, with typically one insertion per cell. (
  • We have developed a web application called iMapper (Insertional Mutagenesis Mapping and Analysis Tool) for the efficient analysis of insertion site sequence reads against vertebrate and invertebrate Ensembl genomes. (
  • Genome-wide LORE1 retrotransposon mutagenesis and high-throughput insertion detection in Lotus japonicus. (
  • We show that active transposon mutagenesis drives ILC formation and analysis of common insertion sites in SB-induced tumors identified a mutually exclusive group of four genes, of which three are frequently aberrated in human ILCs. (
  • High-throughput sequencing of genomic DNA from schistosome populations exposed to MLV mapped widespread and random insertion of transgenes throughout the genome, along each of the autosomes and sex chromosomes, validating the utility of this approach for insertional mutagenesis. (
  • Slingshot represents a flexible and potent system for genome-wide transposon-mediated mutagenesis with many potential applications. (
  • Here, we describe a protocol for the identification of pathogenicity and virulence genes through random insertional mutagenesis using the fungal wilt pathogen Verticillium dahliae as an example for the protocol. (
  • We propose that selective autophagy safeguards genome integrity against excessive insertional mutagenesis caused during nutrient starvation by transposable elements in eukaryotic cells. (
  • While the disruption of normal gene function by transposable elements upon integration into exonic regions is obvious, their post-insertional effects on gene expression have not received much attention. (
  • Retroviral insertional mutagenesis (IM) screens provide one of the most efficient tools to identify genes involved in tumorigenesis and from there the specific oncogenic pathways involved. (
  • The authors concluded that AAV2 is a DNA virus associated with oncogenic insertional mutagenesis in human HCC. (
  • Transposons have been used successfully in lower organisms and plants for insertional mutagenesis, but until activation of the Sleeping Beauty (SB) transposon system, there was no indication of active DNA-based transposons in vertebrates. (
  • The tendency of Ac-Ds transposons to reinsert near the donor site can be used to target both insertional and deletional mutagenesis, but efficient exploitation of this property requires a library of mapped marked donor sites distributed in the genome. (
  • Insertional mutagenesis approaches use oncoretroviruses or transposons to trigger cancer in mice by widespread integration into the cellular genome and activation of oncogenes near the integration site. (
  • To try to find the events that conspire to cause acute myeloid leukaemia, the team studied the same mice using a technique called ' insertional mutagenesis ', in which tagged DNA is inserted into the mouse genome. (
  • Eye abnormality in T27aT15 mice results from random insertional mutagenesis of the transgene as it was only observed in one line. (
  • Secondly, we employed a Sleeping Beauty (SB)-based insertional mutagenesis screen in conditional Cdh1 knockout mice to identify novel genes and pathways involved in the development of ILCs. (
  • A few studies have found increased incidence of hepatocellular carcinomas (HCCs) in mice injected with AAV vectors as newborns whereas no evidence of insertional mutagenesis and cancer were observed in adult mice, in dogs, and in nonhuman primates. (
  • To identify genes capable of cooperating with HER2/neu to fully transform mammary epithelial cells, we used an insertional mutagenesis screen on cells isolated from wild-type neu expressing mice and identified the E3 ligase HACE1 as a HER2 cooperative tumor suppressor gene. (
  • Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice. (
  • We have developed a self-inactivating PiggyBac transposon system for tamoxifen inducible insertional mutagenesis from a stably integrated chromosomal donor. (
  • Thus, we have used a powerful tool for mutagenesis of mammalian cells to reveal an aspect of the complex regulation of NFκB-dependent signaling. (
  • High-throughput mutagenesis of the mammalian genome is a powerful means to facilitate analysis of gene function. (
  • This review highlights the current understanding of the post-insertional effects of TEs on mammalian gene function with a heavy focus on human TEs. (
  • This paper add important information of the controversial issue of AAV and insertional carcinogenesis and will affect development and design of gene therapy trials with AAV vectors. (
  • Two different retroviral vectors, both containing wild type Murine Leukaemia Virus LTRs, were also assayed for insertional mutagenesis in the Bcl15 target cell. (
  • γ-Retroviral vectors (γRVs), which are commonly used in gene therapy, can trigger oncogenesis by insertional mutagenesis. (
  • To identify genes and signaling pathways that initiate Neurofibromatosis type 1 (NF1) neurofibromas, we used unbiased insertional mutagenesis screening, mouse models, and molecular analyses. (
  • Sibirny, A. 2007-08-30 00:00:00 Random insertional mutagenesis is an efficient tool for studying molecular mechanisms of many genetically determined processes. (
  • An improved variant of this method is REMI (Restriction Enzyme Mediated Integration) mutagenesis. (
  • The mechanisms of REMI mutagenesis are surveyed with special reference to yeast Saccharomyces cerevisiae. (
  • Restriction enzyme-mediated integration (REMI) mutagenesis was used to tag genes required for toxin biosynthesis. (
  • Different insertional mutagens have been successfully used to reveal new cancer genes. (
  • In this review, we describe the various insertional mutagens focusing on their advantages/limitations, and we discuss the new and promising tools that will improve the insertional mutagenesis screens of the future. (
  • Mutagenesis that is not random can be used to clone DNA, investigate the effects of mutagens, and engineer proteins. (
  • Insertional inactivation is a technique used in recombinant DNA engineering where a plasmid (such as pBR322) is used to disable expression of a gene. (
  • Site-specific, insertional inactivation of incA in Chlamydia trachomatis using a group II intron. (
  • The mTn5-Nm (containing P tac - nptll -T trpA ) and mTn5-Gm (containing gusA -P tac - nptll -T trpA ) and mTn5-GGm (containing gusA -P tac /P aacCl - aacCl -T trpA ) can be used for transcription signal localization or insertional inactivation. (
  • This document discusses the factors contributing to genotoxicity of vector integration, the strategies to reduce the risk associated to insertional mutagenesis and the assays to evaluate vector oncogenesis at the pre-clinical and clinical level. (
  • The mouse cystic fibrosis transmembrane conductance regulator gene was disrupted in embryonal stem cells using an insertional gene targeting vector. (
  • An important element of vector design is the selection and evaluation of promoter-enhancer elements with sufficient strength to drive reliable immune reconstitution, but minimal propensity for enhancer-mediated insertional mutagenesis. (
  • Retrovirus vector-mediated insertional mutagenesis (IM) can lead to serious cancerous risks in gene therapy. (
  • Though we find a similar rate of insertional mutagenesis, different mechanisms seem to be responsible for transforming Baf3/Bcl15 target cells with a lentiviral compared to a retroviral vector in this mutagenesis assay. (
  • Linear Amplification Mediated PCR for the retrieval of lentiviral vector integration sites from tumors induced by insertional mutagenesis. (
  • Here we describe the adaption and exploitation of LAM-PCR for the retrieval of lentiviral vector (LV) integrations from oligoclonal hepatocellular carcinomas induced by LV-based insertional mutagenesis (4). (
  • There is an example of an insertional mutagenesis event caused by a retrotransposon in the human genome where it causes Fukuyama-type muscular dystrophy. (
  • Selective autophagy regulates insertional mutagenesis by the Ty1 retrotransposon in Saccharomyces cerevisiae. (
  • In lactococci, the study of chromosomal genes and their regulation is limited by the lack of an efficient transposon mutagenesis system. (
  • ATMT-based approaches such as random insertional mutagenesis and targeted knockout are widely used for gene functional analysis in plant-pathogen interactions. (
  • An alternative strategy for insertional mutagenesis has been used in vertebrate animals to find genes that cause cancer. (
  • Our present study provides a basis for the development of efficient transposon-mediated insertional mutagenesis in a vertebrate. (
  • We have combined the highly active SB transposon with gene-trapping technology to demonstrate that transposon traps can be used for insertional mutagenesis screens in vertebrates. (
  • Efficient insertional mutagenesis in lactococci and other gram-positive bacteria. (
  • Because many viruses integrate their own genomes into the genomes of their host cells in order to replicate, mutagenesis caused by viral infections is a fairly common occurrence. (
  • Investigator-driven insertional mutagenesis in vertebrates has relied on retroviral insertions or selection of low-frequency integration of naked DNA in ES cell lines. (
  • Retroviral insertional mutagenesis: the making of a receptor-oncogene. (
  • The gene trap approach has been successfully employed for mutagenesis in vitro and in vivo, and the retrovirus can serve as a useful tag for the identification of the disrupted gene ( 9 , 16 , 21 ). (
  • Use of aminoglycoside 3' adenyltransferase as a selection marker for Chlamydia trachomatis intron-mutagenesis and in vivo intron stability. (
  • Since 2013, development of the CRISPR/Cas9 technology, based on a prokaryotic viral defense system, has allowed for the editing or mutagenesis of a genome in vivo. (
  • Insertional mutagenesis is the phenomenon by which an exogenous DNA sequence integrates within the genome of a host organism. (
  • Furthermore, the approach generates a higher percentage of transformants containing just a single-copy of the integrated T-DNA at random chromosomal sites in the fungal genome allowing for random insertional mutagenesis studies (Mulliuns et al. (
  • A novel method for random mutagenesis of targeted chromosomal regions in Pseudomona aeruginosa was developed. (
  • Recent technical advances have broadened the available approaches for disrupting gene function in a cell population prior to screening, from chemical and insertional mutagenesis to RNA interference, and, most recently, CRISPR mediated genome editing. (
  • To understand mechanisms underlying docetaxel resistance in breast cancer, we used an insertional mutagenesis strategy to identify proteins whose overexpression confers resistance. (
  • Application of this protocol to oligoclonal tumors induced by insertional mutagenesis may optimize the overall retrieval of integrations which are responsible for the cellular transformation and minimize the detection of integrations that occurred in the tumor stroma or in the parenchymal nontumoral cells that can be collected within the tumoral mass together with neoplastic cells. (
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  • The process of disruption or alteration can either be targeted specifically as in the case of gene silencing or homologous recombination or can rely on non-targeted random disruptions (e.g., chemical mutagenesis, transposon mediated mutagenesis) followed by screening a library of individuals for lesions at a specific location. (
  • These findings provide the first description of wide-scale, random insertional mutagenesis of chromosomes and of germline transmission of a transgene in schistosomes. (
  • Site-directed mutagenesis has proved useful in situations that random mutagenesis is not. (
  • however, while random mutagenesis can produce a change in single nucleotides, it does not offer much control as to which nucleotide is being changed. (
  • Our goal was to further probe the structural determinants of arrestin function by using a combined insertional mutagenesis and immunochemical approach. (
  • Results from this analysis can be applied to help identify other important determinants of this Important biological phenomenon and could assist planning of large-scale mutagenesis efforts. (
  • Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. (
  • We have constructed a series of derivatives of the omega interposon [Prentki and Krisch, Gene 29 (1984) 303-313] that can be used for in vitro insertional mutagenesis. (
  • Retroviral insertional mutagenesis in murine mammary cancer. (
  • We are attempting to identify cellular oncogenes activated in mammary tumours by using the mouse mammary tumour virus (MMTV) as an insertional mutagen. (
  • Analysis of HIV-2 Vpx by Modeling and Insertional Mutagenesis" by Lisa A. Mahnke, Michael Belshan et al. (
  • Gene trapping in embryonic stem cells (ESCs) is the most widely used form of-insertional mutagenesis in mammals. (
  • Despite this, the download insertional of twenty-eight arts and the nutritionist of proteins to take their weather Is Written an poor English and embryonic truth. (
  • Extensive analyses failed to implicate insertional mutagenesis or gammac overexpression as the underlying mechanism. (