Mutagenesis: Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.Mutagenesis, Site-Directed: Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.Mutagenesis, Insertional: 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.Molecular Sequence Data: 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.Amino Acid Sequence: 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.Mutation: 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.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Models, Molecular: Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.Escherichia coli: 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.Amino Acid Substitution: 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.Structure-Activity Relationship: 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.Sequence Homology, Amino Acid: The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.DNA Transposable Elements: 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.Recombinant Proteins: Proteins prepared by recombinant DNA technology.Plasmids: 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.Protein Structure, Tertiary: 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.Cloning, Molecular: 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.Bacterial Proteins: Proteins found in any species of bacterium.Kinetics: The rate dynamics in chemical or physical systems.Protein Conformation: 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).Protein Binding: 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.Cysteine: A thiol-containing non-essential amino acid that is oxidized to form CYSTINE.Protein Structure, Secondary: 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.Sequence Alignment: 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.Genes, Bacterial: The functional hereditary units of BACTERIA.Point Mutation: 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.Mutagens: 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.Ethylnitrosourea: A nitrosourea compound with alkylating, carcinogenic, and mutagenic properties.Substrate Specificity: A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.Catalysis: The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction.Alanine: 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.DNA Primers: 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.Cell Line: Established cell cultures that have the potential to propagate indefinitely.SOS Response (Genetics): 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.Histidine: An essential amino acid that is required for the production of HISTAMINE.Conserved Sequence: 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.DNA Mutational Analysis: Biochemical identification of mutational changes in a nucleotide sequence.Catalytic Domain: The region of an enzyme that interacts with its substrate to cause the enzymatic reaction.Escherichia coli Proteins: Proteins obtained from ESCHERICHIA COLI.Crystallography, X-Ray: The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Genetic Complementation Test: 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.Transfection: 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.Restriction Mapping: Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.Ethyl Methanesulfonate: An antineoplastic agent with alkylating properties. It also acts as a mutagen by damaging DNA and is used experimentally for that effect.DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.Sequence Deletion: Deletion of sequences of nucleic acids from the genetic material of an individual.Recombinant Fusion Proteins: 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.Protein Engineering: 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.Oligodeoxyribonucleotides: 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.Lysine: An essential amino acid. It is often added to animal feed.Phenotype: The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.Directed Molecular Evolution: 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.Aspartic Acid: 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.DNA: 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).Amino Acid Motifs: 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.Enzyme Stability: 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.Mutant Proteins: Proteins produced from GENES that have acquired MUTATIONS.Promoter Regions, Genetic: 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.Ultraviolet Rays: 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.Saccharomyces cerevisiae: 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.Transcription, Genetic: The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.Asparagine: 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)Cricetinae: A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.Gene Expression Regulation, Bacterial: Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria.Genetic Vectors: 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.DNA-Binding Proteins: 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.Serine: 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.COS Cells: 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).)Polymerase Chain Reaction: 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.DNA Repair: 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-Directed DNA Polymerase: 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.Arginine: An essential amino acid that is physiologically active in the L-form.Circular Dichroism: 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)Transcription Factors: Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.Sequence Analysis, DNA: A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.Dimerization: The process by which two molecules of the same chemical composition form a condensation product or polymer.Ligands: 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)Tryptophan: 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.Gene Deletion: 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.CHO Cells: 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.Cercopithecus aethiops: 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.Codon: 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, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.Mutagenicity Tests: Tests of chemical substances and physical agents for mutagenic potential. They include microbial, insect, mammalian cell, and whole animal tests.Amino Acids: 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.Operon: 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.Hydrogen-Ion Concentration: 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)Glycosylation: 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.Gene Library: 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.Membrane Proteins: 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.Recombination, Genetic: 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.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.DNA Damage: 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.Transformation, Bacterial: 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.Hydrolysis: The process of cleaving a chemical compound by the addition of a molecule of water.Transformation, Genetic: 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.Methylnitronitrosoguanidine: A nitrosoguanidine derivative with potent mutagenic and carcinogenic properties.Genetic Engineering: 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.Hydrogen Bonding: 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.Biocatalysis: The facilitation of biochemical reactions with the aid of naturally occurring catalysts such as ENZYMES.DNA, Complementary: 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.Consensus Sequence: 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.Alleles: Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.Chromosome Mapping: Any method used for determining the location of and relative distances between genes on a chromosome.DNA Replication: The process by which a DNA molecule is duplicated.Rec A Recombinases: 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.Tyrosine: A non-essential amino acid. In animals it is synthesized from PHENYLALANINE. It is also the precursor of EPINEPHRINE; THYROID HORMONES; and melanin.Mutation, Missense: 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)Glutamic Acid: A non-essential amino acid naturally occurring in the L-form. Glutamic acid is the most common excitatory neurotransmitter in the CENTRAL NERVOUS SYSTEM.Temperature: 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.Protein Processing, Post-Translational: 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.Oligonucleotides: 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)Sequence Homology, Nucleic Acid: 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.Glycine: 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.Virulence: 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.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Phenylalanine: An essential aromatic amino acid that is a precursor of MELANIN; DOPAMINE; noradrenalin (NOREPINEPHRINE), and THYROXINE.Disulfides: Chemical groups containing the covalent disulfide bonds -S-S-. The sulfur atoms can be bound to inorganic or organic moieties.Protein Folding: Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.Protein Structure, Quaternary: The characteristic 3-dimensional shape and arrangement of multimeric proteins (aggregates of more than one polypeptide chain).Nucleic Acid Conformation: The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.Transposases: 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.Saccharomyces cerevisiae Proteins: 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.Oligonucleotide Probes: 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.Frameshift Mutation: 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.Alkylating Agents: 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.Open Reading Frames: 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).Phosphorylation: The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.Genes, Reporter: 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.HeLa Cells: 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.Viral Proteins: Proteins found in any species of virus.Molecular Structure: 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.Threonine: 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.Genetic Techniques: Chromosomal, biochemical, intracellular, and other methods used in the study of genetics.Lac Operon: 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.Gene Expression: The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.Fungal Proteins: Proteins found in any species of fungus.Serine Endopeptidases: Any member of the group of ENDOPEPTIDASES containing at the active site a serine residue involved in catalysis.Genes: 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.Models, Chemical: Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment.Oxidation-Reduction: 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).Blotting, Western: 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.Hypoxanthine Phosphoribosyltransferase: 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 184.108.40.206.Binding, Competitive: 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.Cricetulus: 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.Gene Expression Regulation: 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.Repressor Proteins: 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.Leucine: An essential branched-chain amino acid important for hemoglobin formation.Glutamine: 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.Bacillus subtilis: A species of gram-positive bacteria that is a common soil and water saprophyte.beta-Galactosidase: 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.Peptides: 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, Cultured: 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.Models, Genetic: 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.Gene Targeting: 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.Protein Multimerization: The assembly of the QUATERNARY PROTEIN STRUCTURE of multimeric proteins (MULTIPROTEIN COMPLEXES) from their composite PROTEIN SUBUNITS.Proline: 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, Messenger: 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.Oxidoreductases: 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)Chromosomes, Bacterial: Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.Magnetic Resonance Spectroscopy: 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).HEK293 Cells: A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5.Thermodynamics: 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)Genes, Fungal: The functional hereditary units of FUNGI.Xenopus laevis: 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.Models, Biological: 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.Protein Subunits: 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.Structural Homology, Protein: 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 Transport Proteins: 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.Blotting, Southern: 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, Suppressor: 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.Virus Replication: 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.Suppression, Genetic: 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).Isoleucine: 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 Interaction Domains and Motifs: 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.Peptide Mapping: 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.Mesylates: Organic salts or esters of methanesulfonic acid.Genes, Lethal: 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.GuanineModels, Structural: A representation, generally small in scale, to show the structure, construction, or appearance of something. (From Random House Unabridged Dictionary, 2d ed)DNA, Recombinant: 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.Macromolecular Substances: 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.Biological Transport: 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.Protein Biosynthesis: The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.Multigene Family: 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)DNA, Viral: Deoxyribonucleic acid that makes up the genetic material of viruses.Signal Transduction: 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.Enzyme Activation: 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.Peptide Fragments: Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.Valine: 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.Species Specificity: 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.Nuclear Proteins: 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.Static Electricity: The accumulation of an electric charge on a objectChromosome Deletion: Actual loss of portion of a chromosome.Hydrophobic and Hydrophilic Interactions: The thermodynamic interaction between a substance and WATER.Transcriptional Activation: Processes that stimulate the GENETIC TRANSCRIPTION of a gene or set of genes.Salmonella typhimurium: A serotype of Salmonella enterica that is a frequent agent of Salmonella gastroenteritis in humans. It also causes PARATYPHOID FEVER.Bacterial Outer Membrane Proteins: Proteins isolated from the outer membrane of Gram-negative bacteria.Oocytes: 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).Streptomyces: 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.
VEGF is required for growth and survival in neonatal mice. (1/14345)We employed two independent approaches to inactivate the angiogenic protein VEGF in newborn mice: inducible, Cre-loxP- mediated gene targeting, or administration of mFlt(1-3)-IgG, a soluble VEGF receptor chimeric protein. Partial inhibition of VEGF achieved by inducible gene targeting resulted in increased mortality, stunted body growth and impaired organ development, most notably of the liver. Administration of mFlt(1-3)-IgG, which achieves a higher degree of VEGF inhibition, resulted in nearly complete growth arrest and lethality. Ultrastructural analysis documented alterations in endothelial and other cell types. Histological and biochemical changes consistent with liver and renal failure were observed. Endothelial cells isolated from the liver of mFlt(1-3)-IgG-treated neonates demonstrated an increased apoptotic index, indicating that VEGF is required not only for proliferation but also for survival of endothelial cells. However, such treatment resulted in less significant alterations as the animal matured, and the dependence on VEGF was eventually lost some time after the fourth postnatal week. Administration of mFlt(1-3)-IgG to juvenile mice failed to induce apoptosis in liver endothelial cells. Thus, VEGF is essential for growth and survival in early postnatal life. However, in the fully developed animal, VEGF is likely to be involved primarily in active angiogenesis processes such as corpus luteum development. (+info)
Membrane-tethered Drosophila Armadillo cannot transduce Wingless signal on its own. (2/14345)Drosophila Armadillo and its vertebrate homolog beta-catenin are key effectors of Wingless/Wnt signaling. In the current model, Wingless/Wnt signal stabilizes Armadillo/beta-catenin, which then accumulates in nuclei and binds TCF/LEF family proteins, forming bipartite transcription factors which activate transcription of Wingless/Wnt responsive genes. This model was recently challenged. Overexpression in Xenopus of membrane-tethered beta-catenin or its paralog plakoglobin activates Wnt signaling, suggesting that nuclear localization of Armadillo/beta-catenin is not essential for signaling. Tethered plakoglobin or beta-catenin might signal on their own or might act indirectly by elevating levels of endogenous beta-catenin. We tested these hypotheses in Drosophila by removing endogenous Armadillo. We generated a series of mutant Armadillo proteins with altered intracellular localizations, and expressed these in wild-type and armadillo mutant backgrounds. We found that membrane-tethered Armadillo cannot signal on its own; however it can function in adherens junctions. We also created mutant forms of Armadillo carrying heterologous nuclear localization or nuclear export signals. Although these signals alter the subcellular localization of Arm when overexpressed in Xenopus, in Drosophila they have little effect on localization and only subtle effects on signaling. This supports a model in which Armadillo's nuclear localization is key for signaling, but in which Armadillo intracellular localization is controlled by the availability and affinity of its binding partners. (+info)
Bone resorption induced by parathyroid hormone is strikingly diminished in collagenase-resistant mutant mice. (3/14345)Parathyroid hormone (PTH) stimulates bone resorption by acting directly on osteoblasts/stromal cells and then indirectly to increase differentiation and function of osteoclasts. PTH acting on osteoblasts/stromal cells increases collagenase gene transcription and synthesis. To assess the role of collagenase in the bone resorptive actions of PTH, we used mice homozygous (r/r) for a targeted mutation (r) in Col1a1 that are resistant to collagenase cleavage of type I collagen. Human PTH(1-34) was injected subcutaneously over the hemicalvariae in wild-type (+/+) or r/r mice four times daily for three days. Osteoclast numbers, the size of the bone marrow spaces and periosteal proliferation were increased in calvariae from PTH-treated +/+ mice, whereas in r/r mice, PTH-induced bone resorption responses were minimal. The r/r mice were not resistant to other skeletal effects of PTH because abundant interstitial collagenase mRNA was detected in the calvarial periosteum of PTH-treated, but not vehicle-treated, r/r and +/+ mice. Calcemic responses, 0.5-10 hours after intraperitoneal injection of PTH, were blunted in r/r mice versus +/+ mice. Thus, collagenase cleavage of type I collagen is necessary for PTH induction of osteoclastic bone resorption. (+info)
DMPK dosage alterations result in atrioventricular conduction abnormalities in a mouse myotonic dystrophy model. (4/14345)Myotonic dystrophy (DM) is the most common form of muscular dystrophy and is caused by expansion of a CTG trinucleotide repeat on human chromosome 19. Patients with DM develop atrioventricular conduction disturbances, the principal cardiac manifestation of this disease. The etiology of the pathophysiological changes observed in DM has yet to be resolved. Haploinsufficiency of myotonic dystrophy protein kinase (DMPK), DM locus-associated homeodomain protein (DMAHP) and/or titration of RNA-binding proteins by expanded CUG sequences have been hypothesized to underlie the multi-system defects observed in DM. Using an in vivo murine electrophysiology study, we show that cardiac conduction is exquisitely sensitive to DMPK gene dosage. DMPK-/- mice develop cardiac conduction defects which include first-, second-, and third-degree atrioventricular (A-V) block. Our results demonstrate that the A-V node and the His-Purkinje regions of the conduction system are specifically compromised by DMPK loss. Importantly, DMPK+/- mice develop first-degree heart block, a conduction defect strikingly similar to that observed in DM patients. These results demonstrate that DMPK dosage is a critical element modulating cardiac conduction integrity and conclusively link haploinsufficiency of DMPK with cardiac disease in myotonic dystrophy. (+info)
Predicting insecticide resistance: mutagenesis, selection and response. (5/14345)Strategies to manage resistance to a particular insecticide have usually been devised after resistance has evolved. If it were possible to predict likely resistance mechanisms to novel insecticides before they evolved in the field, it might be feasible to have programmes that manage susceptibility. With this approach in mind, single-gene variants of the Australian sheep blowfly, Lucilia cuprina, resistant to dieldrin, diazinon and malathion, were selected in the laboratory after mutagenesis of susceptible strains. The genetic and molecular bases of resistance in these variants were identical to those that had previously evolved in natural populations. Given this predictive capacity for known resistances, the approach was extended to anticipate possible mechanisms of resistance to cyromazine, an insecticide to which L. cuprina populations remain susceptible after almost 20 years of exposure. Analysis of the laboratory-generated resistant variants provides an explanation for this observation. The variants show low levels of resistance and a selective advantage over susceptibles for only a limited concentration range. These results are discussed in the context of the choice of insecticides for control purposes and of delivery strategies to minimize the evolution of resistance. (+info)
Accelerated accumulation of somatic mutations in mice deficient in the nucleotide excision repair gene XPA. (6/14345)Inheritable mutations in nucleotide excision repair (NER) genes cause cancer-prone human disorders, such as xeroderma pigmentosum, which are also characterized by symptoms of accelerated ageing. To study the impact of NER deficiency on mutation accumulation in vivo, mutant frequencies have been determined in liver and brain of 2-16 month old NER deficient XPA-/-, lacZ hybrid mice. While mutant frequencies in liver of 2-month old XPA-/-, lacZ mice were comparable to XPA+/-, lacZ and the lacZ parental strain animals, by 4 months of age mutant frequencies in the XPA-deficient mice were significantly increased by a factor of two and increased further until the age of 16 months. In brain, mutant frequencies were not found to increase with age. These results show that a deficiency in the NER gene XPA causes an accelerated accumulation of somatic mutations in liver but not in brain. This is in keeping with a higher incidence of spontaneous liver tumors reported earlier for XPA-/- mice after about 15 months of age. (+info)
Inward rectification in KATP channels: a pH switch in the pore. (7/14345)Inward-rectifier potassium channels (Kir channels) stabilize the resting membrane potential and set a threshold for excitation in many types of cell. This function arises from voltage-dependent rectification of these channels due to blockage by intracellular polyamines. In all Kir channels studied to date, the voltage-dependence of rectification is either strong or weak. Here we show that in cardiac as well as in cloned KATP channels (Kir6.2 + sulfonylurea receptor) polyamine-mediated rectification is not fixed but changes with intracellular pH in the physiological range: inward-rectification is prominent at basic pH, while at acidic pH rectification is very weak. The pH-dependence of polyamine block is specific for KATP as shown in experiments with other Kir channels. Systematic mutagenesis revealed a titratable C-terminal histidine residue (H216) in Kir6.2 to be the structural determinant, and electrostatic interaction between this residue and polyamines was shown to be the molecular mechanism underlying pH-dependent rectification. This pH-dependent block of KATP channels may represent a novel and direct link between excitation and intracellular pH. (+info)
The Gab1 PH domain is required for localization of Gab1 at sites of cell-cell contact and epithelial morphogenesis downstream from the met receptor tyrosine kinase. (8/14345)Stimulation of the hepatocyte growth factor (HGF) receptor tyrosine kinase, Met, induces mitogenesis, motility, invasion, and branching tubulogenesis of epithelial and endothelial cell lines in culture. We have previously shown that Gab1 is the major phosphorylated protein following stimulation of the Met receptor in epithelial cells that undergo a morphogenic program in response to HGF. Gab1 is a member of the family of IRS-1-like multisubstrate docking proteins and, like IRS-1, contains an amino-terminal pleckstrin homology domain, in addition to multiple tyrosine residues that are potential binding sites for proteins that contain SH2 or PTB domains. Following stimulation of epithelial cells with HGF, Gab1 associates with phosphatidylinositol 3-kinase and the tyrosine phosphatase SHP2. Met receptor mutants that are impaired in their association with Gab1 fail to induce branching tubulogenesis. Overexpression of Gab1 rescues the Met-dependent tubulogenic response in these cell lines. The ability of Gab1 to promote tubulogenesis is dependent on its pleckstrin homology domain. Whereas the wild-type Gab1 protein is localized to areas of cell-cell contact, a Gab1 protein lacking the pleckstrin homology domain is localized predominantly in the cytoplasm. Localization of Gab1 to areas of cell-cell contact is inhibited by LY294002, demonstrating that phosphatidylinositol 3-kinase activity is required. These data show that Gab1 is an important mediator of branching tubulogenesis downstream from the Met receptor and identify phosphatidylinositol 3-kinase and the Gab1 pleckstrin homology domain as crucial for subcellular localization of Gab1 and biological responses. (+info)
Plasmid-based one-pot saturation mutagenesis<...
TY - JOUR. T1 - Plasmid-based one-pot saturation mutagenesis. AU - Wrenbeck,Emily E.. AU - Klesmith,Justin R.. AU - Stapleton,James A.. AU - Adeniran,Adebola. AU - Tyo,Keith E.J.. AU - Whitehead,Timothy A.. PY - 2016/11/1. Y1 - 2016/11/1. N2 - Deep mutational scanning is a foundational tool for addressing the functional consequences of large numbers of mutants, but a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped plasmid dsDNA. The method can be used to produce comprehensive or single- or multi-site saturation mutagenesis libraries.. AB - Deep mutational scanning is a foundational tool for addressing the functional consequences of large numbers of mutants, but a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Here we present nicking mutagenesis, a robust, ...
Empirical Complexities in the Genetic Foundations of Lethal Mutagenesis | Genetics
From population genetics theory, elevating the mutation rate of a large population should progressively reduce average fitness. If the fitness decline is large enough, the population will go extinct in a process known as lethal mutagenesis. Lethal mutagenesis has been endorsed in the virology literature as a promising approach to viral treatment, and several in vitro studies have forced viral extinction with high doses of mutagenic drugs. Yet only one empirical study has tested the genetic models underlying lethal mutagenesis, and the theory failed on even a qualitative level. Here we provide a new level of analysis of lethal mutagenesis by developing and evaluating models specifically tailored to empirical systems that may be used to test the theory. We first quantify a bias in the estimation of a critical parameter and consider whether that bias underlies the previously observed lack of concordance between theory and experiment. We then consider a seemingly ideal protocol that avoids this ...
When defense turns into attack: Antiviral cytidine deaminases linked to somatic mutagenesis in HPV-associated cancer. - PubMed...
Mutagenesis | BaseClear B.V.
Site-directed mutagenesis of a sequence to make specific, targeted changes to double stranded DNA. Based on optimised protocols, BaseGene is able to create mutations in virtually every DNA fragment. This is a valuable tool for studying DNA or protein structure and function. Site-directed or random mutagenesis can be applied to any DNA-fragment cloned into a plasmid vector. We can help you to introduce a range of mutations, varying from point mutations to deletions or insertions.. ...
PCR MUTAGENESIS PROBLEM - Molecular Biology
If you have plan to use commercial kits, another choice for short way for your research is use of mutagenesis service company. I realized the mutagenesis cost using commercial kit is never cheap! My estimate is $200 ~ 250 per one mutagenesis reaction using kit (one kit rexn., $20 + purified two primers, $150 + three clone sequencing, $30 + subcloning reagent, $30 : SM, media, agar plate, buffer, tube, tips --- and time). Besides, purchasing kit for just one or two mutant generation is money wasting because remaining kit reagents are useless. Recent biotech companies provide rapid and precise mutagenesis service in affordable prices. Some people in our lab use Mutagenex Inc. (USA) and I found other companies offering in low price ...
Lyckeby - EU Court of Justice ruling
Mutations occur when a DNA strand is damaged and the cells own repair mechanism mends the damage so that the end result is different from what it looked like to start with. The mutations thus arise through the organisms own system; regardless of whether it is a spontaneous mutation, a mutation with traditional methods, to which the EU court refers to having shown a history of safe use, or new improved methods of refinement. Since spontaneous mutations as well as mutagenesis through traditional methods have demonstrated safe use, it cannot be interpreted otherwise than that all mutagenesis involving a breach in the DNA strand and mutation induced through the organisms repair system do not cause any safety problems. It is not possible to distinguish in a crop the technique used for mutagenesis, and thus the exception in the GMO Directive must apply regardless of the mutagenesis technique used. In the judgment, it has been decided primarily to review recital 17 in combination with Annex IB of ...
Weird problem in in vitro mutagenesis: post #1
Weird problem in in vitro mutagenesis - posted in Molecular Biology: Hi, Iam doing invitro mutagenesis for a 5.2Kb plasmid using stratagene kit. I did not get any results with my plasmid, so checked the control in the kit. Unfortunately the control is not working. I have tried many times, but got no colonies. The conditions used were according to the instructions given by the manufacturer. 95 C 50 sec 95 C 50 seconds 55 C 1 min 68 C 5 minute x 18 times 68 C 5 minutes. I even tried addin...
Mutagenesis of the mouse Bhlhb4 gene. A: Gene targeting | Open-i
Mutagenesis | Vastenhouw Lab
Yesterday Cindy presented the results of her Masters rotation project, which she carried out in our lab over the past couple of months. During the rotation, she worked with Lennart to develop and optimize a new gene excision protocol. Not all details can be revealed yet, but it looks like we can cut long sequences surprisingly early and efficiently!. As was also recognized in Cindys exam presentation, she completed a considerable amount of work. She also became a lively part of the lab - so that time passed too fast, again, and her next rotation period is coming up. We wish the best of luck and exciting results!. ...
In-vitro Mutagenesis | Protocol
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Unraveling innate immunity using large scale N-ethyl-N-nitrosourea mutagenesis<...
TY - JOUR. T1 - Unraveling innate immunity using large scale N-ethyl-N-nitrosourea mutagenesis. AU - Hoebe, Kasper. AU - Beutler, B.. PY - 2005/5. Y1 - 2005/5. N2 - With the mouse genome almost entirely sequenced and readily accessible to all who wish to examine it, the challenge across most biological disciplines now lies in the decipherment of gene and protein function rather than in the realm of gene identification per se. In the field of innate immunity, forward genetic methods have repeatedly been applied to identify key sensors, adapters, and effector molecules. However, most spontaneous mutations that affect innate immune function have been mapped and cloned, and the need for new monogenic phenotypes has been felt evermore keenly. N-Ethyl-N-nitrosourea (ENU) mutagenesis is an efficient tool for the creation of aberrant monogenic innate immune response phenotypes. In this review, we will discuss the potential of the forward genetic approach and ENU mutagenesis to identify new genes and new ...
A systematic, genome-wide, phenotype-driven mutagenesis programme for gene function studies in the mouse. - Department of...
As the human genome project approaches completion, the challenge for mammalian geneticists is to develop approaches for the systematic determination of mammalian gene function. Mouse mutagenesis will be a key element of studies of gene function. Phenotype-driven approaches using the chemical mutagen ethylnitrosourea (ENU) represent a potentially efficient route for the generation of large numbers of mutant mice that can be screened for novel phenotypes. The advantage of this approach is that, in assessing gene function, no a priori assumptions are made about the genes involved in any pathway. Phenotype-driven mutagenesis is thus an effective method for the identification of novel genes and pathways. We have undertaken a genome-wide, phenotype-driven screen for dominant mutations in the mouse. We generated and screened over 26,000 mice, and recovered some 500 new mouse mutants. Our work, along with the programme reported in the accompanying paper, has led to a substantial increase in the mouse mutant
Large-scale mutagenesis of the mouse to understand the genetic bases of nervous system structure and function<...
TY - JOUR. T1 - Large-scale mutagenesis of the mouse to understand the genetic bases of nervous system structure and function. AU - Goldowitz, Dan. AU - Frankel, Wayne N.. AU - Takahashi, Joseph S.. AU - Holtz-Vitaterna, Martha. AU - Bult, Carol. AU - Kibbe, Warren A.. AU - Snoddy, Jay. AU - Li, Yanxia. AU - Pretel, Stephanie. AU - Yates, Jeana. AU - Swanson, Douglas J.. PY - 2004/12/20. Y1 - 2004/12/20. N2 - N-ethyl-N-nitrosourea (ENU) mutagenesis is presented as a powerful approach to developing models for human disease. The efforts of three NIH Mutagenesis Centers established for the detection of neuroscience-related phenotypes are described. Each center has developed an extensive panel of phenotype screens that assess nervous system structure and function. In particular, these screens focus on complex behavioral traits from drug and alcohol responses to circadian rhythms to epilepsy. Each of these centers has developed a bioinformatics infrastructure to track the extensive number of ...
The relation between repair of DNA and radiation and chemical mutagenesis in Saccharomyces cerevisiae<...
TY - JOUR. T1 - The relation between repair of DNA and radiation and chemical mutagenesis in Saccharomyces cerevisiae. AU - Prakash, Louise. PY - 1976. Y1 - 1976. N2 - The effect of various genes involved in DNA repair functions on radiation and chemical mutagenesis in Escherichia coli is discussed and compared to similar studies done yeast. Results of the effect of various genes conferring radiation-sensitivity on mutation induction in yeast are presented and related to current ideas of mutagenesis.. AB - The effect of various genes involved in DNA repair functions on radiation and chemical mutagenesis in Escherichia coli is discussed and compared to similar studies done yeast. Results of the effect of various genes conferring radiation-sensitivity on mutation induction in yeast are presented and related to current ideas of mutagenesis.. UR - http://www.scopus.com/inward/record.url?scp=0017042140&partnerID=8YFLogxK. UR - ...
Rxivist: Genome-wide chemical mutagenesis screens allow unbiased saturation of the cancer genome and identification of drug...
Drug resistance is an almost inevitable consequence of cancer therapy and ultimately proves fatal for the majority of patients. In many cases this is the consequence of specific gene mutations that have the potential to be targeted and re-sensitize the tumor. The means therefore to saturate the genome with point mutations and that avoids chromosome or nucleotide sequence context bias would open the door to identify all possible drug resistance mutations in cancer models. Here we describe such a method for elucidating drug resistance mechanisms using genome-wide chemical mutagenesis allied to next-generation sequencing. We show that chemically mutagenizing the genome of cancer cells dramatically increases the number of drug-resistant clones and allows the detection of both known and novel drug resistance mutations. We have developed an efficient computational process that allows for the rapid identification of involved pathways and druggable targets. Such a priori knowledge would greatly empower ...
GENETAILOR SITE DIRECTED MUTAGENESIS PDF
mutagenesis - Drug Discovery Opinion
The concept of lethal mutagenesis has been developed as a means of curing viral infections and has also been used to explain the action of some antiviral drugs. Although mutation is the basis for adaptation and survival, especially in the presence of antiviral drugs, most mutations are detrimental and the theory of lethal mutagenesis holds that an infecting population can be pushed to extinction by an overwhelmingly high mutation rate. Chemical mutagens have been used to increase error rates in a number of RNA viruses including HIV-1 and HCV and have been found to significantly reduce viral titres and, in some cases, achieve extinction. For example, the antiviral activity of the ribonucleoside analogue 5-azacytidine (5-AZC) against HIV-1 has been attributed primarily to an increase in mutant frequency consistent with lethal mutagenesis caused by incorporation of 5-AZC into viral DNA.. A team of researchers from the University of Texas at Austin have now raised serious concerns about the ...
Mutagenesis: Site-specific - eLS - El-Gewely - Wiley Online Library
Site-specific mutagenesis techniques are aimed at the precise substitution, insertion or deletion of any coding sequence in vitro. More recently, however, such precise alterations are also being developed for in vivo gene/genome modifications. These techniques are revolutionizing our understanding of the genetic and molecular mechanisms in several biological systems, which could lead to the development of new enzymes, therapeutics as well as improved agricultural applications. ...
In Vitro Mutagenesis Protocols (Methods... | WHSmith Books
Site-directed mutagenesis - experimental considerations | NEB
Environmental and Molecular Mutagenesis - Volume 53, Issue 3 - April 2012 - Wiley Online Library
International round-robin study on the Ames fluctuation test (pages 185-197). G. Reifferscheid, H.M. Maes, B. Allner, J. Badurova, S. Belkin, K. Bluhm, F. Brauer, J. Bressling, S. Domeneghetti, T. Elad, S. Flückiger-Isler, H.J. Grummt, R. Gürtler, A. Hecht, M.B. Heringa, H. Hollert, S. Huber, M. Kramer, A. Magdeburg, H.T. Ratte, R. Sauerborn-Klobucar, A. Sokolowski, P. Soldan, T. Smital, D. Stalter, P. Venier, Chr. Ziemann, J. Zipperle and S. Buchinger. Version of Record online: 4 JAN 2012 , DOI: 10.1002/em.21677. ...
Tools to Study Mammalian Mutagenesis - Stephen Dertinger
"Somatic Mutagenesis with a Sleeping Beauty Transposon System Leads to " by Maura McGrail, Julia M. Hatler et al.
Large-scale sequencing of human cancer genomes and mouse transposon-induced tumors has identified a vast number of genes mutated in different cancers. One of the outstanding challenges in this field is to determine which genes, when mutated, contribute to cellular transformation and tumor progression. To identify new and conserved genes that drive tumorigenesis we have developed a novel cancer model in a distantly related vertebrate species, the zebrafish, Danio rerio. The Sleeping Beauty (SB) T2/Onc transposon system was adapted for somatic mutagenesis in zebrafish. The carp ß-actin promoter was cloned into T2/Onc to create T2/OncZ. Two transgenic zebrafish lines that contain large concatemers of T2/ OncZ were isolated by injection of linear DNA into the zebrafish embryo. The T2/OncZ transposons were mobilized throughout the zebrafish genome from the transgene array by injecting SB11 transposase RNA at the 1-cell stage. Alternatively, the T2/OncZ zebrafish were crossed to a transgenic line that
Fast Mutagenesis System,Fast MultiSite Mutagenesis System,Mutagenesis System,Beijing TransGen B
Chemical mutagenesis and Tilling - Symbiotic Bacteria
Prior to the implementation of more sophisticated genetic techniques, mutagenesis was often performed using simple chemical mutagens including N-methyl-N-nitro-N-nitroso-guanidine (MMNG) and ethyl methane sulfonate (EMS) . The basic approach involves exposing a population of cells to a controlled dose of mutagen, to generate progeny that carry random genetic mutations The application of this technique has traditionally been limited by two important considerations First, it often proves difficult to strike a balance between efficiency and selectivity; exposure to the mutagen must be carefully controlled in order to maximize the frequency of single mutations Second, it should be noted that chemical mutagenesis is random; the experimenter needs to select or identify mutants of interest from a large pool of random mutants. For more detailed information on the background and procedures involved in chemical mutagenesis, we refer the reader to Miller (1992) The basic protocols that have been described ...
Combinatorial mutagenesis and pseudorevertant analysis to charact...: Ingenta Connect
Deletion of the I265-F268 and T271-K277 regions in the large lumenally exposed loop of the CP47 protein are known to lead to a loss of photoautotrophic growth. Here, these regions have been investigated by combinatorial mutagenesis and pseudorevertant mapping. No single amino-acid residue in the I265-F268 region was found to be critical for function, but a large hydrophobic residue at position 267 and preferentially an aromatic residue at position 268 appeared to be required for photoautotrophic growth. Starting from an obligate photoheterotrophic mutant lacking the T271-K277 region, photoautotrophic pseudorevertants were generated with short in-frame tandem repeats near the site of the original deletion, partially or fully restoring the length of the original protein. These pseudorevertants were sensitive to oxygen indicating that the T271-K277 region may provide PS II stability and/or protection against oxygen-dependent photoinactivation. Pseudorevertants with much improved photoautotrophic ...
"Rapid in vivo forward genetic approach for identifying axon death gene" by Lukas J. Neukomm, Thomas C. Burdett et al.
Axons damaged by acute injury, toxic insults, or neurodegenerative diseases execute a poorly defined autodestruction signaling pathway leading to widespread fragmentation and functional loss. Here, we describe an approach to study Wallerian degeneration in the Drosophila L1 wing vein that allows for analysis of axon degenerative phenotypes with single-axon resolution in vivo. This method allows for the axotomy of specific subsets of axons followed by examination of progressive axonal degeneration and debris clearance alongside uninjured control axons. We developed new Flippase (FLP) reagents using proneural gene promoters to drive FLP expression very early in neural lineages. These tools allow for the production of mosaic clone populations with high efficiency in sensory neurons in the wing. We describe a collection of lines optimized for forward genetic mosaic screens using MARCM (mosaic analysis with a repressible cell marker; i.e., GFP-labeled, homozygous mutant) on all major autosomal arms (
Iterative Saturation Mutagenesis (ISM): A Powerful Approach to Engineer Proteins by Systematically Simulating Darwinian...
Author: Acevedo Rocha, Carlos Guillermo et al.; Genre: Book Chapter; Published in Print: 2014-08-31; Keywords: deletion mutations - directed evolution - gene variant libraries - insertion mutations - mutagenesis; Title: Iterative Saturation Mutagenesis (ISM): A Powerful Approach to Engineer Proteins by Systematically Simulating Darwinian Evolution
Type 2 diabetes mellitus (T2DM) is the leading disorder of metabolism worldwide, and it is predicted to affect more than 330 million people over the next quarter century, generating increasing social and economic burdens. At the pathophysiological level, T2DM arises from a combination of resistance to the hormone insulin and insufficient production of insulin by the endocrine pancreas. T2DM is a polygenic disorder, and many of the genes involved are still unknown. N-ethyl-N-nitrosourea (ENU)-mutagenesis is a forward genetic approach that can be used to identify genes that can be mutated to cause a phenotype of interest. In an effort to identify genes that play a role in regulating blood glucose levels we performed an ENU screen in mice. Recessive and dominant screens were performed, and we identified fourteen lines of mice with evidence for heritable hyperglycemia. Glucokinase (Gck) was a candidate gene, and so we sequenced for Gck mutations in most of the fourteen lines. We identified two lines ...
"Supporting conditional mouse mutagenesis with a comprehensive cre char" by Caleb S Heffner, C Herbert Pratt et al.
Full realization of the value of the loxP-flanked alleles generated by the International Knockout Mouse Consortium will require a large set of well-characterized cre-driver lines. However, many cre driver lines display excision activity beyond the intended tissue or cell type, and these data are frequently unavailable to the potential user. Here we describe a high-throughput pipeline to extend characterization of cre driver lines to document excision activity in a wide range of tissues at multiple time points and disseminate these data to the scientific community. Our results show that the majority of cre strains exhibit some degree of unreported recombinase activity. In addition, we observe frequent mosaicism, inconsistent activity and parent-of-origin effects. Together, these results highlight the importance of deep characterization of cre strains, and provide the scientific community with a critical resource for cre strain information.
Mouse Mutagenesis Program Produces New Disease Models | BioWorld
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Mutagenesis legal definition of mutagenesis
ProxiMAX randomization: a new technology for non-degenerate saturation mutagenesis of contiguous codons | Biochemical Society...
A) DNA sequence analysis of a library encoded by six cycles of codon addition, optimized to provide unbiased representation. Six cycles of ProxiMAX randomization were undertaken as described in Supplementary Figure S2 (at http://www.biochemsoctrans.org/bst/041/bst0411189add.htm), using right-handed hairpins (Supplementary Table S3 at http://www.biochemsoctrans.org/bst/041/bst0411189add.htm) as donors and an amplicon of pUC19 as the acceptor, with optimized mixtures of 18 codons adjusted to reflect the sequence bias illustrated in Supplementary Figure S2(B). The resulting library was analysed by DNA sequencing, using a MiSeq DNA sequencer according to the manufacturers instructions. Data represent the analysis of 286684 sequences of the correct length, which represented 79.9% of the entire library. Bars represent the frequency of each codon from each cycle of saturation mutagenesis. The broken line depicts the target representation for each codon. Further analysis of the library can be found in ...
Site-Directed Mutagenesis - Cloning and Mutagenesis - GENEWIZ
GENEWIZ can increase your research productivity by performing your time-consuming site-directed mutagenesis projects efficiently and cost-effectively. Our customized mutagenesis services provide a fail-safe approach to obtain mutant constructs quickly, with 100% accuracy, thus eliminating the possibility of undesired mutations in your gene.. ...
Genetic Modifier Screens in Mice - Current Protocols
Lethal mutagenesis of HIV with mutagenic nucleoside analogs | PNAS
Our results indicate that growth of HIV in tissue culture in the presence of 5-OH-dC results in the loss of the HIV population and in the accumulation of G → A substitutions. In seven of nine serial experiments, a precipitous decline in viral infectivity occurred over serial passage. This result contrasts with simultaneous incubations carried out in the absence of a nucleoside analog; in a total of 28 control cultures in which the supernatant was serially transferred from 7 to as many as 34 times, the replication of HIV was never abolished, nor was the viral titer diminished by more than 90%. Furthermore, abolishment of HIV titer was not observed with nine other deoxynucleoside analogs so far tested. We detected 97 new 5-OH-dC-induced mutations in 53,000 nucleotides (bottom of Table 3). Assuming that these mutations were evenly distributed throughout an HIV genome containing 10,000 nucleotides, then each proviral DNA obtained immediately prior to lethality contains approximately 18 ...
How long will it take to finish the mutagenesis project? - GenScript Help Center
Gene site saturated mutagenesis
Environmental Carcinogenesis and Mutagenesis - Peter Stambrook
Ethidium Bromide mutagenesis
Chromosomal Mutagenesis | Shondra M. Pruett-Miller | Springer
The Mechanism of Nucleotide Excision Repair-Mediated UV-Induced Mutagenesis in Non-Proliferating Cells | Genetics
Following the irradiation of non-dividing yeast cells with ultraviolet (UV) light, most induced mutations are inherited by both daughter cells, indicating that complementary changes are introduced into both strands of duplex DNA prior to replication. Early analyses demonstrated that such "two-strand" mutations depend on functional nucleotide excision repair (NER), but the molecular mechanism of this unique type of mutagenesis has not been further explored. In the experiments reported here, an ade2 adeX colony-color system was used to examine the genetic control of UV-induced mutagenesis in non-dividing cultures of Saccharomyces cerevisiae. We confirmed a strong suppression of two-strand mutagenesis in NER-deficient backgrounds, and demonstrated that neither mismatch repair nor interstrand crosslink repair affects the production these mutations. By contrast, proteins involved in the error-prone bypass of DNA damage (Rev3, Rev1, PCNA, Rad18, Pol32 and Rad5) and in the early steps of the DNA-damage ...
Beyond knockouts: cre resources for conditional mutagenesis<...
TY - JOUR. T1 - Beyond knockouts: cre resources for conditional mutagenesis. AU - Murray, Stephen A. AU - Eppig, Janan. AU - Smedley, Damian. AU - Simpson, Elizabeth M. AU - Rosenthal, Nadia Alicia. PY - 2012. Y1 - 2012. N2 - With the effort of the International Phenotyping Consortium to produce thousands of strains with conditional potential gathering steam, there is growing recognition that it must be supported by a rich toolbox of cre driver strains. The approaches to build cre strains have evolved in both sophistication and reliability, replacing first-generation strains with tools that can target individual cell populations with incredible precision and specificity. The modest set of cre drivers generated by individual labs over the past 15+ years is now growing rapidly, thanks to a number of large-scale projects to produce new cre strains for the community. The power of this growing resource, however, depends upon the proper deep characterization of strain function, as even the best ...
Insertional mutagenesis - definition of insertional mutagenesis by The Free Dictionary
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...
Physical Mutagenesis in Giant Reed (Arundo Donax L.) and Phenotypic and Genomic Characterization of Mutagenized Clones
ABSTRACT Giant reed (Arundo donaxL.) is a C3 perennial, warm-season, rhizomatous grass of emerging interest for bioenergy and biomass-derivatives production, and for phytoremediation. We developed and applied a mutagenesis protocol based on γ-irradiation of in-vitro cell cultures from which regenerants were obtained. Based on a radio-sensitivity test, the irradiation dose reducing to 50% the number of regenerants per callus (RD50) was estimated at 35 Gy. A large mutagenic experiment was carried out by irradiating a total of 3,120 calli with approx. 1x, 1.5x and 2x RD50. A total of 1,004 regenerants from irradiated calli were hardened in pots and transplanted to the field. Approx. 10% of field grown clones showed remarkable morphological aberrations including dwarfism, altered tillering, abnormal inflorescence, leaf variegation and others, which were tested for stability over generations. Clone lethality reached 0.4%. Our results show for the first time that physical mutagenesis can efficiently ...
Spontaneous mutagenesis in Csbm/mOgg1−/− mice is attenuated by dietary resveratrol : Carcinogenesis - oi
Oxidative DNA modifications such as 7,8-dihydro-8-oxoguanine (8-oxoG) are generated endogenously in apparently all living cells. The defect of the repair of 8-oxoG in Csbm/mOgg1−/− mice results in elevated basal levels of these lesions and increased frequencies of spontaneous mutations, which initiate tumorigenesis in the liver if cell proliferation is stimulated. Here, we describe that the phytoalexin resveratrol, applied either for 7 days per gavage (100 mg/kg body wt) or for 3-9 months in the diet (0.04% ad libitum), reduces the endogenous oxidative DNA base damage in the livers of the Csbm/mOgg1−/− mice by 20-30% (P , 0.01). A small but consistent effect is also observed in the wild-type animals. The spontaneous mutation frequencies determined in the lacI gene of BigBlue® Csbm/mOgg1−/− mice are concomitantly reduced by resveratrol to similar extents. Mechanistically, the protection is caused by an induction of the antioxidant defense system since (i) hepatocytes isolated from ...
ZFIN Publication: Muto et al., 2005
The visual system converts the distribution and wavelengths of photons entering the eye into patterns of neuronal activity, which then drive motor and endocrine behavioral responses. The gene products important for visual processing by a living and behaving vertebrate animal have not been identified in an unbiased fashion. Likewise, the genes that affect development of the nervous system to shape visual function later in life are largely unknown. Here we have set out to close this gap in our understanding by using a forward genetic approach in zebrafish. Moving stimuli evoke two innate reflexes in zebrafish larvae, the optomotor and the optokinetic response, providing two rapid and quantitative tests to assess visual function in wild-type (WT) and mutant animals. These behavioral assays were used in a high-throughput screen, encompassing over half a million fish. In almost 2,000 F2 families mutagenized with ethylnitrosourea, we discovered 53 recessive mutations in 41 genes. These new mutations ...
Strategies for Mutagenesis and Gene Cloning Using Transposon Tagging and T-DNA Insertional Mutagenesis, Annual Review of Plant...
A thermostable variant of P. aeruginosa cold-adapted LipC obtained by rational design and saturation mutagenesis :: MPG.PuRe
Author: Cesarini, Silvia et al.; Genre: Journal Article; Published in Print: 2012-12; Keywords: Cold-adapted lipase|br/|Rational design|br/|Saturation mutagenesis|br/|Enzyme improvement|br/|Pseudomonas; Title: A thermostable variant of P. aeruginosa cold-adapted LipC obtained by rational|br/|design and saturation mutagenesis
Appearance and properties of L-sorbose-utilizing mutants of Candida albicans obtained on a selective plate • Research -...
Difference between revisions of "IGEM:UW" - OpenWetWare
Our team is planning to develop two connected devices. The first device will introduce mutations to defined region of plasmid in E. coli cells (level of mutations in the rest of bacterial genome should be near to background). We will try to achieve this using human protein AID (Activation Induced Deamidase). It acts on single stranded DNA causing C → U substitutions. This activity causes point mutations and activation of various DNA repair systems potentially including error-prone DNA repair systems. Site-specific in vivo DNA mutagenesis device can be used to produce proteins with new activities. In this case proper selection pressure must be applied. Our second device will be used to create proper selection conditions. It will use beta-lactamase split into two complementing domains anchored on E. coli cell surface and culture medium containing ampicillin to promote survival of clones expressing desired phenotype and ensure death of others. One beta-lactamase domain will be bound to cell ...
Favipiravir Elicits Antiviral Mutagenesis During Virus Replication in Vivo
Lethal mutagenesis has emerged as a novel potential therapeutic approach to treat viral infections. Several studies have demonstrated that increases in the high mutation rates inherent to RNA viruses lead to viral extinction in cell culture, but evidence during infections in vivo is limited. In this …
Random mutation is antithetical to natural selection
Citation Machine: Mutation Research Fundamental And Molecular Mechanisms Of Mutagenesis format citation generator for raw rata
Describe how you would use site-directed mutagenesis to, Ask an Expert
DSpace at EWHA: Identification of novel genes responsible For multiple tolerance By transposon mutagenesis In Saccharomyces...
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 ...
Tag: random mutation | Evolution News
Environmental Mutagenesis & Genomics Society
Towards therapy-induced lethal HIV mutagenesis - [email protected]
8.10. Summary · Functional Programming in OCaml
But mutability does have its uses. I/O is fundamentally about mutation. And some data structures (like arrays and hash tables) cannot be implemented as efficiently without mutability.. Mutability thus offers great power, but with great power comes great responsibility. Try not to abuse your new-found power!. ...
Used in insertional mutagenesis *Insertional mutagenesis uses the features of a TE to insert a sequence. In most cases this is ... Mechanisms of mutagenesis. TEs are mutagens and their movements are often the causes of genetic disease. They can damage ... Insertional mutagenesis uses the features of a TE to insert a sequence. In most cases this is used to remove a DNA sequence or ... Used for analysis of gene expression and protein functioning in signature-tagging mutagenesis. *This analytical tool allows ...
Clyde A. Hutchison III
Site-directed mutagenesis. In 1971, Clyde Hutchison and Marshall Edgell showed that it is possible to produce mutants ... "The Development of Site-directed Mutagenesis by Michael Smith" (PDF). Journal of Biological Chemistry. 281 (39).. ... Clyde A. Hutchison III is an American biochemist and microbiologist notable for his research on site-directed mutagenesis and ... Hutchison, C.A. III; Swanstrom, R. & Loeb, D.D. (1991). "Complete Mutagenesis of Protein Coding Domains". Methods in Enzymology ...
P Raychaudhury; Basu, Ashis K. (2011). "Genetic requirement for mutagenesis of the G[8,5-Me]T cross-link in Escherichia coli: ... Friedberg EC, Walker GC, Siede W, Wood RD, Schultz RA, Ellenberger T. (2006). DNA Repair and Mutagenesis, part 3. ASM Press. ... Paromita Raychaudhury and Ashis Basu studied the toxicity and mutagenesis of the same lesion in Escherichia coli by replicating ... Volkert MR (1988). "Adaptive response of Escherichia coli to alkylation damage". Environmental and Molecular Mutagenesis. 11 (2 ...
Mutagenesis. 4 (5): 349-54. doi:10.1093/mutage/4.5.349. PMID 2687628. Brookman KW, Lamerdin JE, Thelen MP, Hwang M, Reardon JT ... DNA Repair and Mutagenesis. ASM Press. ISBN 1555813194. "Entrez Gene: ERCC4 excision repair cross-complementing rodent repair ... Environmental and Molecular Mutagenesis. 51 (6): 520-6. doi:10.1002/em.20569. PMC 3017513 . PMID 20658645. Sargent RG, Rolig RL ... Environmental and Molecular Mutagenesis. 51 (6): 567-81. doi:10.1002/em.20583. PMID 20658648. Clauson C, Schärer OD, ...
Mutagenesis. 26 (1): 125-132. doi:10.1093/mutage/geq052. ISSN 0267-8357. PMID 21164193. Savage, John R. K. (1988-01-01). "A ... Mutagenesis. 22 (3): 195-200. doi:10.1093/mutage/gem002. ISSN 0267-8357. Bandana Ganguly, Bani (1993-08-01). "Cell division, ... Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. 41 (2-3): 321-331. doi:10.1016/0027-5107(76)90105-6. ...
Friedberg, EC; Walker, GC; Siede, W; Wood, RD; Schultz, RA; Ellenberger, T (2006). DNA Repair and Mutagenesis. ASM Press. ISBN ... Mutagenesis. 4 (5): 349-54. doi:10.1093/mutage/4.5.349. PMID 2687628. Sgouros J, Gaillard PH, Wood RD (Mar 1999). "A ... "Evolution and mutagenesis of the mammalian excision repair gene ERCC-1". Nucleic Acids Research. 16 (12): 5305-22. doi:10.1093/ ... Environmental and Molecular Mutagenesis. 51 (6): 567-81. doi:10.1002/em.20583. PMID 20658648. Jaspers NG, Raams A, Silengo MC, ...
Gel electrophoresis of nucleic acids
Herpes simplex virus
Mutagenesis. 15: 261-269. doi:10.1093/mutage/15.3.261. Hosaka, S; et al. (1980). "Effect of Cytochalasin B and Colchicine on ... Mutagenesis: 743-747. "Genotoxicity: The Micronucleus Test". Crios. Retrieved 19 March 2013. Fenech, M; Kirsch-Volders, M; ... Mutagenesis. 26: 125-132. doi:10.1093/mutage/geq052. PMID 21164193. Umegaki, K; Fenech, M (2000). "Cytokinesis-Block ...
In-vitro Mutagenesis | Protocol
PCR MUTAGENESIS PROBLEM - Molecular Biology
PCR MUTAGENESIS PROBLEM - SiteDirected Mutagenesis - Problem (Jul/21/2005 ). Im doing invitro pcr mutagenesis DpnI digestion ... I realized the mutagenesis cost using commercial kit is never cheap! My estimate is $200 ~ 250 per one mutagenesis reaction ... chimeric gene generation and multiple-site mutagenesis. Especially, they show low mutagenesis efficiency (below 40%) for genes ... The mutagenesis efficiency is close to 100%. Presently, the overlap extension method, megaprimer method, Quick Change method ( ...
Mutagenesis | Vastenhouw Lab
Mutagenesis | BaseClear B.V.
Plasmid-based one-pot saturation mutagenesis<...
Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped ... Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped ... Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped ... Here we present nicking mutagenesis, a robust, single-day, one-pot saturation mutagenesis method performed on routinely prepped ...
Lyckeby - EU Court of Justice ruling
Mutagenesis has been used as a breeding method for over 70 years and more than 3000 varieties have mutagenesis in their lineage ... Without mutagenesis as a refining tool, we would today face even greater global challenges having lower crop yields and ... The European Court of Justice recently came up with a ruling on how to interpret the use of mutagenesis in an existing GMO ... 1)mutagenesis,. (2)cell fusion (including protoplast fusion) of plant cells of organisms which can exchange geneticmaterial ...
Radiation Mutagenesis | Encyclopedia.com
Source for information on Radiation Mutagenesis: World of Microbiology and Immunology dictionary. ... Radiation mutagenesis Mutations are caused by DNA damage and genetic alterations that may occur spontaneously at a very low ... Radiation mutagenesis. Mutations are caused by DNA damage and genetic alterations that may occur spontaneously at a very low ... www.encyclopedia.com/science/encyclopedias-almanacs-transcripts-and-maps/radiation-mutagenesis ...
Ethidium bromide (EtBr ) is widely used in molecular biology as a specific dye for staining DNA: the intercalated molecule fluoresces on exposure to ultraviolet light. EtBr may be incorporated into electrophoretic gels, or the gel may be stained after running. In Bio2250 labs, care should always be taken not to expose oneself to either gels or buffer containing EtBr. ...
Chemical Carcinogenesis and Mutagenesis II | SpringerLink
I have been privileged to witness and participate in the great growth of knowledge on chemical carcinogenesis and mutagenesis ... Carcinom DNA Mutagene Onkogene Viren cancer cancer research carcinogenesis cellular differentiation mutagen mutagenesis ... I have been privileged to witness and participate in the great growth of knowledge on chemical carcinogenesis and mutagenesis ... cinogenesis and mutagenesis, including their recent and fruitful union with viral oncology. I feel very optimistic about the ...
... Zophonias O. Jonsson zjons at vetbio.unizh.ch Fri Feb 3 05:05:18 EST 1995 *Previous message: PCR mutagenesis ... 21 (1991), p. 6052, Random mutagenesis of gene-sized DNA molecules by use of PCR with Taq DNA polymerase Johan H. Spee et al., ... 3 (1993), p. 777-778, Efficient random mutagenesis method with adjustable mutation frequency by use of PCR and dITP Marcel F. ... I am interested in finding a protocol for PCR mutagenesis that will , incorporate random mismatches while replicating. Any ...
... Philip Batterham philip_batterham at MUWAYF.UNIMELB.EDU.AU Wed Apr 5 21:07:16 EST 1995 *Previous message: ... ABOUT MUTAGENESIS Hello. My name is Kris Freebairn. I am currently doing a pHD in the department of Genetics at the University ... To do this I am using mutagenesis. So far, I have used EMS which has been quite successful. As EMS has a strong bias towards GC ...
Chromosomal Mutagenesis | Shondra M. Pruett-Miller | Springer
Includes all new cutting-edge methods and protocols for chromosomal mutagenesis research Provides step-by-step detail essential ... Chromosomal Mutagenesis. Editors. * Shondra M. Pruett-Miller Series Title. Methods in Molecular Biology. Series Volume. 1239. ... Authoritative and fully updated, Chromosomal Mutagenesis, Second Edition aims to help speed scientific discovery and aid in the ... Since the first edition, major advances and discoveries have made chromosomal mutagenesis a widely used technique and one that ...
Lethal Mutagenesis of Bacteria | Genetics
1998) and are therefore not considered candidates for lethal mutagenesis.. Lethal mutagenesis has not been proposed as a ... Surprisingly, models of lethal mutagenesis indicate that bacteria may be candidates for lethal mutagenesis. In contrast to ... Graci, J. D., D. A. Harki, V. S. Korneeva, J. P. Edathil, K. Too et al., 2007 Lethal mutagenesis of poliovirus mediated by a ... Graci, J. D., K. Too, E. D. Smidansky, J. P. Edathil, E. W. Barr et al., 2008 Lethal mutagenesis of picornaviruses with n-6- ...
Ethidium Bromide mutagenesis
MUTAGENESIS | Kirkus Reviews
In the 27th century, after devastating wars, Earth is rediscovering its long-lost, far-flung star colonies. The most remote of these, planet Anu, may be the repository of food grasses long extinct on Earth. But when an Earth expedition arrives, geneticist Mattie Manan encounters a culture so severely patriarchal that it refuses to negotiate at all if she is allowed off the ship. Though apparently primitive, this same culture employs ``losos, speechless, upright human-animal hybrids, to do the work and tend the repressed womenfolk. Suspecting matters even more seriously amiss, Mattie escapes from the ship, then heads for the mysterious Eastcountry accompanied by inhibited natives Elizabeth, a gifted artist, and Erin, a mechanical-mathematical genius, whose talents gradually blossom now that their ``fathers are no longer present. After various adventures, Mattie is captured by the Eastcountry geneticist-whiz ``elves, who plan to eliminate the female sex altogether, leaving only male genetic supermen
tag :: mutagenesis, | BibSonomy
AcidAminoBindingBinding,ComplexElectronHeme,Motifs,Mutagenesis,MutantNeisseriaNitrites,ProteinProteins,Sequence,Sites,Subunits ... Analysis,BacterialBacterial,BaseChainChromosomeChromosomes,Cloning,Data,Elements,GeneticMapping,MolecularMolecular,Mutagenesis, ... SignalAcidAminoAnimalsBindingCHOCricetinaeHumansLeucine/*chemistry/geneticsMutagenesis,MutationNucleicPointRepetitiveSequences, ... AcidAcids,AminoAnalysis,BacterialBacterial,Capsules,Data,Genetic,Hexosyltransferases,MolecularMutagenesis,NeisseriaPlasmids, ...
Video Mutagenesis - Videomaker
QUT - Plasma mutagenesis
Plasma mutagenesis is a novel microbial mutation breeding technology developed in recent years, which uses a combination of UV ... Mutagenesis is an important technique in microbial engineering whereby DNA mutations are deliberately introduced into the ... There are two main approaches, i.e., rational design and random mutagenesis. Rational design is highly useful when there is ... You will use plasma mutagenesis to create a library of mutants with the aim of improving microbial growth on relevant ...
Mutagenesis - Wikipedia
Directed mutagenesis Site-directed mutagenesis PCR mutagenesis Insertional mutagenesis Signature tagged mutagenesis Transposon ... Early methods of mutagenesis produced entirely random mutations; however, later methods of mutagenesis may produce site- ... Mutagenesis as a science was developed based on work done by Hermann Muller, Charlotte Auerbach and J. M. Robson in the first ... Mutagenesis may also arise as a result of the presence of environmental mutagens that induce changes to the DNA. The mechanism ...
Transgenesis and Targeted Mutagenesis in Immunology - 1st Edition
Purchase Transgenesis and Targeted Mutagenesis in Immunology - 1st Edition. Print Book & E-Book. ISBN 9780121057602, ... Transgenesis and Targeted Mutagenesis in Immunology 1st Edition. 0 star rating Write a review ... qu:The present volume represents a concise and complete guide to transgenesis and targeted mutagenesis in laboratory mice. ... This breakthrough laboratory guide provides a complete study of transgenesis and targeted mutagenesis in laboratory mice that ...
tagging mutagenesis in yeast??
... Eckhard Boles boles at uni-duesseldorf.de Thu Oct 24 11:37:31 EST 1996 *Previous message: ... Hi, I would like to perform a random mutagenesis with a specific S. cerevisiae mutant strain selecting for a positive phenotype ... Can anyone provide me with information about tagging mutagenesis systems in yeast, e.g. inducible Ty transposable elements on a ...
Cassette mutagenesis - Wikipedia
Cassette mutagenesis is a type of site-directed mutagenesis that uses a short, double-stranded oligonucleotide sequence (gene ... Unlike many site directed mutagenesis methods, cassette mutagenesis also does not involve primer extension by DNA polymerase. ... Compared to mutagenesis methods that requires the synthesis of double stranded DNA using a single stranded template (1-30% in ... Kegler-Ebo, D. M.; Docktor, C. M.; Dimaio, D (1994). "Codon cassette mutagenesis: A general method to insert or replace ...
Chemical Carcinogenesis and Mutagenesis II | Colin S. Cooper | Springer
I have been privileged to witness and participate in the great growth of knowledge on chemical carcinogenesis and mutagenesis ... Chemical Carcinogenesis and Mutagenesis II. Editors: Cooper, Colin S., Grover, Philip L. (Eds.) ... I have been privileged to witness and participate in the great growth of knowledge on chemical carcinogenesis and mutagenesis ... cinogenesis and mutagenesis, including their recent and fruitful union with viral oncology. I feel very optimistic about the ...
Translesion DNA synthesis and mutagenesis in eukaryotes. - PubMed - NCBI
Translesion DNA synthesis and mutagenesis in eukaryotes.. Sale JE1.. Author information. 1. Medical Research Council Laboratory ... This, in turn, stimulates PCNA ubiquitination, recruitment of Pol η, and Phase II mutagenesis at A/T base pairs. ... DNA and how they are regulated to balance their ability to replicate DNA lesions with the risk of undesirable mutagenesis. It ...
Mutagenesis: Site-specific - eLS - El-Gewely - Wiley Online Library
Lethal mutagenesis of HIV with mutagenic nucleoside analogs | PNAS
Lethal mutagenesis of HIV with mutagenic nucleoside analogs. Lawrence A. Loeb, John M. Essigmann, Farhad Kazazi, Jue Zhang, ... Lethal mutagenesis of HIV with mutagenic nucleoside analogs. Lawrence A. Loeb, John M. Essigmann, Farhad Kazazi, Jue Zhang, ... 1998) in Oxidative DNA Damage and Mutagenesis, eds Nickoloff J A, Hoekstra M F (Humana, Totowa, NJ), 1, pp 1-18. ... Lethal mutagenesis of HIV with mutagenic nucleoside analogs. Lawrence A. Loeb, John M. Essigmann, Farhad Kazazi, Jue Zhang, ...
METHYL METHANESULFONATE MUTAGENESIS IN BACTERIOPHAGE T4 | Genetics
... directed mutagenesis (SDM) aims to introduceprecise alterations in any coding or noncoding deoxyribonucleic acid (DNA) sequence ... a) Overlap extension method for site‐directed mutagenesis. (b) SLiCE‐mediated PCR‐based site‐directed mutagenesis (SLiP site‐ ... El‐Gewely MR (1991) Oligonucleotide and multisite directed mutagenesis. In: El‐Gewely MR (ed.) Site Directed Mutagenesis and ... Keywords: codon optimisation; mutation efficiency; site‐saturation mutagenesis; random mutagenesis; N‐end rule; synthetic ...
Mutagenesis & Cloning | Agilent
Our large portfolio features site-directed mutagenesis and cloning products for any of your downstream applications. ... Mutagenesis and Cloning products, including the latest in next generation kits and vectors to streamline your workflow, with ... Extensive range of classic and next generation Mutagenesis and Cloning products. Agilent Mutagenesis and Cloning portfolio ... Competent Cells for Mutagenesis. , Competent Cells for Routine Cloning. Enzymes, Inhibitors & Kits Enzymes, inhibitors and kits ...
Evidence for APOBEC3B mutagenesis in multiple human cancers. - PubMed - NCBI
Insertional mutagenesis - definition of insertional mutagenesis by The Free Dictionary
insertional mutagenesis synonyms, insertional mutagenesis pronunciation, insertional mutagenesis translation, English ... Noun 1. insertional mutagenesis - a mutation caused by the insertion of exogenous DNA into a genome genetic science, genetics ... insertional mutagenesis. Also found in: Thesaurus, Medical, Wikipedia. ThesaurusAntonymsRelated WordsSynonymsLegend: ... insertional mutagenesis - (genetics) a mutation caused by the insertion of exogenous DNA into a genome. genetic science, ...
Gene Discovery by MMTV Mediated Insertional Mutagenesis | SpringerLink
In: Dupuy A., Largaespada D. (eds) Insertional Mutagenesis Strategies in Cancer Genetics. Springer, New York, NY. * DOI https ... 2008). Large-scale mutagenesis in p19(ARF)- and p53-deficient mice identifies cancer genes and their collaborative networks. ... 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 ...
Q5® Site-Directed Mutagenesis Kit | NEB
... site-specific mutagenesis of double-stranded plasmid DNA in less than 2 hours. The kit utilizes the robust Q5 Hot Start High- ... The Q5® Site-Directed Mutagenesis Kit enables rapid, ... Q5® Site-Directed Mutagenesis Kit Quick Protocol (E0554) * Q5® ... DNA Assembly, Cloning and Mutagenesis Kits. Applications: Site Directed Mutagenesis, Site Directed Mutagenesis. * Advantages ... The Q5® Site-Directed Mutagenesis Kit enables rapid, site-specific mutagenesis of double-stranded plasmid DNA in less than 2 ...
Site-directed mutagenesis - experimental considerations | NEB
Site-directed mutagenesis - experimental considerations. Return to Site Directed Mutagenesis Another important aspect to ... Home Applications DNA Amplification, PCR and qPCR Site Directed Mutagenesis Site-directed mutagenesis - experimental ... Incomplete primer synthesis can lead to errors at the mutagenesis site. As synthesis errors occur more often with larger ... has been developed not only to aid in the design of primers for the Q5 Site-Directed Mutagenesis Kit, but also to provide an ...
Site-directed Mutagenesis from Exon BioSystems
... any number of mutageneses. ExonBio provides the best service with double sequecing to make sure the success of the mutagenesis ... Site-directed Mutagenesis from Exon BioSystems,Any form, any size, ... GPS -M Mutagenesis System from New England Biolabs. 5. GeneTailor Site-Directed Mutagenesis System from Invitrogen. 6. Mouse ... Site-directed mutagenesis-Within 10 bases from Bio S&T. 2. Site-directed DNA Mutagenesis Service from Yorkshire Bioscience Ltd ...
LibrariesMethodInsertionalGenomeGeneRapid and efficient site‐directedEfficiencySystematic MutagenesisChromosomal MutagenesisGeneticMutationsMutantsMediated Mutagenesis ProtocolEscherichiaCarcinogenesisVitroPlasmidProtocolCRISPRSynthesisGenomesLethal mutagenesisProtocolsPolymeraseEfficientProteinApproachesDeletionOligonucleotidePrimerColiRandomMutantPrimersMiceOverlapLaboratoryEnvironmentalReactionOrganismsQuikChangeSiteCloningHeritable
- Deep mutational scanning is a foundational tool for addressing the functional consequences of large numbers of mutants, but a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. (msu.edu)
- The method can be used to produce comprehensive or single- or multi-site saturation mutagenesis libraries. (msu.edu)
- In Chromosomal Mutagenesis , a team of experts provide a variety of chromosomal manipulation techniques, including insertional gene disruptions, gene knockouts, stimulated homologous recombination techniques and other novel tools, for both prokaryotic and eukaryotic organisms, and attempt to expand the genetic toolbox beyond model organisms. (springer.com)
- Can anyone provide me with information about tagging mutagenesis systems in yeast, e.g. inducible Ty transposable elements on a plasmid, which after creating the insertional mutation allow to identify the mutated gene? (bio.net)
- 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. (thefreedictionary.com)
- 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. (thefreedictionary.com)
- Point mutation was found in the vast majority of all patients, insertional mutagenesis in 4 cases. (thefreedictionary.com)
- 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. (thefreedictionary.com)
- The structural genes of adeno-associated virus serotype 2 (AAV2) have been altered by linker insertional mutagenesis in order to define critical components of virion assembly and infectivity. (nih.gov)
- 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. (europa.eu)
- ATMT-based approaches such as random insertional mutagenesis and targeted knockout are widely used for gene functional analysis in plant-pathogen interactions. (wur.nl)
- 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. (wur.nl)
- To date, the generation of C. jejuni mutant libraries has been limited to transposon shuttle mutagenesis ( 4 ) and homologous insertional mutagenesis ( 2 , 16 ). (asm.org)
- We therefore designed and tested an in vivo mariner -based transposon mutagenesis system for the production of random insertional mutants of C. jejuni . (asm.org)
- However, lethal mutagenesis has been attempted only with viruses that have RNA genomes, whose intrinsic mutation rates are high enough (∼1 per genome per generation) that they might be pushed over an extinction threshold with only a slight mutational increase. (genetics.org)
- The exceptionally high rate of mutagenesis of RNA viruses ( 11 , 12 ), coupled with the finding that most HIV virions in the blood appear to be nonviable ( 13 ), suggest that the HIV genome is unable to tolerate many additional mutations without a loss of viability. (pnas.org)
- Since 2013, the development of the CRISPR /Cas9 technology, based on a prokaryotic viral defense system, has also allowed for the editing of the genome , and mutagenesis may be performed in vivo with relative ease. (wikipedia.org)
- Global transposon mutagenesis was used to identify nonessential genes in an effort to learn whether the naturally occurring gene complement is a true minimal genome under laboratory growth conditions. (sciencemag.org)
- To evaluate whether Cpf1 can be applied to plant genome editing, we selected Cpf1 from Francisella novicida (FnCpf1), which recognizes a shorter PAM (TTN) within known Cpf1 proteins, and applied it to targeted mutagenesis in tobacco and rice. (nature.com)
- Insect transposons could therefore be used for high frequency insertion mutagenesis of the human genome in functional genomics and high-throughput screening. (biomedcentral.com)
- There are some fundamental differences between genome editing and mutagenesis applied in conventional plant breeding that are important for risk assessment and traceability resp. (testbiotech.org)
- The following table lists some of the differences between genome editing and mutagenesis. (testbiotech.org)
- NAR Vol. 19 No. 21 (1991), p. 6052, Random mutagenesis of gene-sized DNA molecules by use of PCR with Taq DNA polymerase Johan H. Spee et al. (bio.net)
- In the laboratory, however, mutagenesis is a useful technique for generating mutations that allows the functions of genes and gene products to be examined in detail, producing proteins with improved characteristics or novel functions, as well as mutant strains with useful properties. (wikipedia.org)
- Hi, I would like to perform a random mutagenesis with a specific S. cerevisiae mutant strain selecting for a positive phenotype which enables me to identify the corresponding gene. (bio.net)
- Cassette mutagenesis is a type of site-directed mutagenesis that uses a short, double-stranded oligonucleotide sequence (gene cassette) to replace a fragment of target DNA. (wikipedia.org)
- This limitation can be overcome with a technology known variously as gene targeting, targeted mutagenesis, or gene replacement the subject of this section. (jax.org)
- Site-directed mutagenesis is a molecular biology method that is used to make specific and intentional changes to the DNA sequence of a gene and any gene products . (wikipedia.org)
- There are numerous methods for achieving site-directed mutagenesis, but with decreasing costs of oligonucleotide synthesis , artificial gene synthesis is now occasionally used as an alternative to site-directed mutagenesis. (wikipedia.org)
- In this work, they often encounter questions about the likelihood of off-target mutagenesis - unintended genetic mutations introduced by the gene editing process - in their mouse lines. (eurekalert.org)
- Targeted mutagenesis and gene targeting using sequence specific nucleases (SSNs) are powerful strategies used to accelerate molecular breeding of crops. (nature.com)
- Site-directed mutagenesis is the method of choice for altering a gene or vector sequence at a selected location. (genengnews.com)
- However, all of them have shortcomings for applying to the diverse mutagenesis strategies including base substitution, deletion, insertion, chimeric gene generation and multiple-site mutagenesis. (protocol-online.org)
- it is likely that nuclear annexin A1 is involved in DNA replication, especially DNA damage induced gene mutation, since DNA damage induced mutagenesis plays an important role in tumorigenesis. (intechopen.com)
- People have mostly been looking at mutations in the DNA sequence that codes for protein, but in this paper we found that the promoter, the regulatory element of gene expression, is very susceptible to mutagenesis," said Wang, "and this susceptibility is facilitated by head-on transcription and DNA replication . (phys.org)
- This book addresses cutting-edge techniques for researching transposon mutagenesis, an approach for identifying individual gene contributions to the phenotypic characteristics of a particular microorganism. (springer.com)
- Mutagenesis is a fundamental approach in biology to identify gene function, and in plants it may involve using chemicals, ionizing radiation, or specific DNA insertion sequences. (plantphysiol.org)
Rapid and efficient site‐directed2
- Adachi Y and Fukuhara C (2012) TA strategy for rapid and efficient site‐directed mutagenesis. (els.net)
- Rapid and Efficient Site-directed Mutagenesis by the Single-tube Megaprimer PCR Method Protocol Protocol for rapid and efficient site-directed mutagenesis by the single-tube megaprimer PCR method. (molecularstation.com)
- The high efficiency of the mutagenesis means mutants can be screened directly by sequencing. (wikipedia.org)
- Our kits speed up the protocol for performing site-directed or random mutagenesis, with high efficiency and 100% accuracy and reliability. (agilent.com)
- Many approaches have since been developed to improve the efficiency of mutagenesis. (wikipedia.org)
- They also plan to examine the tradeoff between improving the efficiency of Cas9 mutagenesis and improving its precision. (eurekalert.org)
- Today's conventional site-directed mutagenesis methods, first introduced in 1995 (see Strategies 9(1): 3-4 under "Resources"), typically utilize a three-step, one-day method to introduce point mutations, amino acid substitutions, deletions, and small insertions in virtually any double-stranded plasmid template with high rates of efficiency. (genengnews.com)
- The efficiency and ease-of-use of site-directed mutagenesis methods have facilitated engineering of promoter and coding regions of numerous genes. (genengnews.com)
- Looking forward, leading the drive for continued innovation in the area of mutagenesis means embracing novel technologies, which may yield new enhancements in speed, efficiency, fidelity, or throughput. (genengnews.com)
- The GeneArt® Site-Directed Mutagenesis System has been optimized for efficiency and simplicity. (thermofisher.com)
- resulting in a higher mutagenesis efficiency. (thermofisher.com)
- Each GeneArt® Site-Directed Mutagenesis System includes enough reagents for 16 reactions and a control: DNA methylase, S-adenosylmethionine, enzyme mix and buffer, enhancer, 0.5M EDTA, sterile water, pUC19WHITE mutagenesis control, and One Shot® MAX Efficiency DH5α™-T1R chemically competent cells. (thermofisher.com)
- After site-directed mutagenesis, H 1-2 P 3 H 4 P 5-6 H 7-8 (E24D & E83G) with increased homology (91.45%) with dHU and higher activity and catalytic efficiency than the FDA-approved porcine-baboon chimera (PBC) was obtained. (mdpi.com)
- Therefore, development of efficient procedure by rational mutagenesis manipulation of L-asparaginase to increase enzyme efficiency is in high interest. (ukessays.com)
- Great disparities exist between organisms with regard to the relative ease of chromosomal mutagenesis and manipulation. (springer.com)
- Since the first edition, major advances and discoveries have made chromosomal mutagenesis a widely used technique and one that is available to any molecular biology laboratory, and this collection provides detailed protocols, case-studies, and reviews from thought-leaders in the field. (springer.com)
- Authoritative and fully updated, Chromosomal Mutagenesis, Second Edition aims to help speed scientific discovery and aid in the next advances in the field. (springer.com)
- used in the total download chromosomal mutagenesis by P. Brittain: A Life( London: name Press, 2001), course This family is right designed. (sara-jordan.com)
- Watergate download chromosomal mutagenesis came powerful. (sara-jordan.com)
- 27 Words from Rome and Venice am once heavily in one download chromosomal mutagenesis. (sara-jordan.com)
- This, specifically, Lenin remained requirements a download chromosomal mutagenesis in November. (sara-jordan.com)
- It is almost 35,000 women on download chromosomal mutagenesis, both for Equal service almost not as helping reviewers. (sara-jordan.com)
- Comprehensive and cutting-edge, Chomosomal Mutagenesis covers state-of-the-art techniques that are staged to expand, if not revolutionize, genetic analysis in the long neglected and relevant cell types. (springer.com)
- Mutagenesis /mjuːtəˈdʒɛnɪsɪs/ is a process by which the genetic information of an organism is changed, resulting in a mutation. (wikipedia.org)
- In vitro mutagenesis using dut − ung − genetic selection method. (els.net)
- Plant mutagenesis is the induction of new genetic variants, frequently by treatment with chemicals, ionizing radiation, or mobilization of insertion elements. (els.net)
- Mutation Research - Genetic Toxicology and Environmental Mutagenesis (MRGTEM) publishes papers advancing knowledge in the field of genetic toxicology. (elsevier.com)
- The reported research uses a reverse mutagenesis bacteriophage T4D model to quantitatively study the effects of direct current E-fields (DC/E) on a molecular genetic level. (thefreedictionary.com)
- In a report that appears online today in the Proceedings of the National Academy of Sciences , she and her colleagues not only show that this stress-induced mutagenesis occurs in an entirely new assay system, it also is a significant contributor to spontaneous mutation - those changes in the genetic code that are not purposefully caused in the laboratory. (healthcanal.com)
- Authoritative and practical, Microbial Transposon Mutagenesis: Protocols and Applications serves as a valuable reference for scientists seeking to apply transposon mutagenesis to microbial genetic analyses and functionality. (springer.com)
- Genetic studies of Campylobacter jejuni have been limited due to the lack of a transposon mutagenesis method. (asm.org)
- In order to respond to these basic and priority questions p, we are developing novel genetic tools for random mutagenesis in Plasmodium falciparum that would permit mutant production on large scale. (europa.eu)
- In practice, mutagenesis can require many generations before extinction is achieved, allowing the bacterial population to grow to large absolute numbers before the load of deleterious mutations causes the decline. (genetics.org)
- MMS-induced forward mutation is sharply reduced by the mutations px and y , which also reduce ultraviolet, photodynamic and γ-ray mutagenesis and increase killing by all of these agents. (genetics.org)
- Quick Tips - Can I make multiple mutations with the Q5 site-directed mutagenesis kit? (neb.com)
- Early attempts at mutagenesis using radiation or chemical mutagens were non-site-specific, generating random mutations. (wikipedia.org)
- SB mutagenesis provides a rich resource of mutations in potential cancer drivers for cross-comparative analyses with ongoing sequencing efforts in human pancreatic adenocarcinoma. (pnas.org)
- The GeneArt™ site-directed mutagenesis service introduces single or multiple mutations (substitutions, insertions, deletions, or truncations) into existing DNA sequences, quickly and economically. (thermofisher.com)
- In addition to the improved site-directed mutagenesis method described above, a newer approach was developed with the ability to create mutations at multiple sites simultaneously. (genengnews.com)
- It's a resounding confirmation of the regulation of mutagenesis by stress responses, which causes mutations specifically when cells are maladapted to their environment when mutations might allow the cell to adapt," said Rosenberg. (thefreedictionary.com)
- on directed mutagenesis in certain bacteria belittles the importance of natural selection in the evolutionary process particularly because no one expected that beneficial mutations could be induced from within the cell. (thefreedictionary.com)
- Neuroscience-focused mutagenesis and phenotyping facilities established by this RFA are expected to serve as a national resource by producing a bank of mouse strains that harbor a wide range of mutations affecting murine nervous system function and behavior. (nih.gov)
- The GeneArt® Site-Directed Mutagenesis System provides a state-of-the-art, simple, convenient, and highly efficient means to generate mutations on a target construct in vitro in less than three hours. (thermofisher.com)
- I found the several reviews of in vitro mutagenesis work, especially interesting and valuable, but was surprised that chemical mutagens, which have proved so effective in seed treatments, have not proved beneficial to date for the induction of somatic mutations in somaclonal culture systems. (thefreedictionary.com)
- To facilitate these studies, methodologies have been developed to introduce specific mutations that can be directly applied to the mutagenesis of infectious clones (icBAC) using BAC technologies. (mdpi.com)
- Hydroxylamine Mutagenesis of Plasmid DNA Protocol Protocol for hydroxylamine mutagenesis of plasmid DNA is ideal for random mutagenesis of plasmid DNA which is then used in a plasmid shuffle or screen for ts mutants. (molecularstation.com)
- In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI Protocol for in vitro mutagenesis using double-stranded DNA templates. (molecularstation.com)
- Creating mutants with the GeneArt® Site-Directed Mutagenesis System relies on the inherent properties of DNA methylase, high fidelity DNA polymerase, recombination enzymes, and the native McrBC endonuclease of E. coli . (thermofisher.com)
- In vitro mutagenesis , plant regeneration and characterization of mutants via RAPD analysis in Baby's breath Gypsophila paniculata L. (thefreedictionary.com)
- In addition to the excellent transcriptomics, proteomics, metabolomics, and bioinformatics tools being developed for both species by different groups around the world, a sizeable number of mutants have been collected using various mutagenesis approaches. (plantphysiol.org)
- In particular, the lack of an in vivo transposon mutagenesis method for the efficient generation of random mutants of Campylobacter spp. (asm.org)
- This will greatly potentiate the yield of produced mutants since it will allow the expansion of parasite clones that contain the regulatable transposase before mutagenesis activation. (europa.eu)
Mediated Mutagenesis Protocol1
- I have been privileged to witness and participate in the great growth of knowledge on chemical carcinogenesis and mutagenesis since 1939 when I entered graduate school in biochemistry at the University of Wisconsin- Madison. (springer.com)
- The programme was chosen to explore what is currently known about the mechanisms of mutagenesis and carcinogenesis, induced by environmental agents, and the questions regarding the relationship of these two processes. (waterstones.com)
- Site-specific mutagenesis techniques are aimed at the precise substitution, insertion or deletion of any coding sequence in vitro . (wiley.com)
- Site‐directed mutagenesis (SDM) aims to introduce precise alterations in any coding or noncoding deoxyribonucleic acid (DNA) sequence, usually in vitro . (els.net)
- In Vitro Mutagenesis Protocols (Methods. (whsmith.co.uk)
- In vitro mutagenesis and identification of new variants via RAPD markers for improving chrysanthemum morifolium. (thefreedictionary.com)
- In vitro mutagenesis quick method for establishment of solid mutant in Chrysanthemum. (thefreedictionary.com)
- Influence of in vitro mutagenesis on ovule culture and plant regeneration in cotton (Gossypium hirsutum L. (thefreedictionary.com)
- Providing an overview of in vitro mutagenesis, along with in-depth protocols and troubleshooting advice, the guide makes it easy to carry out efficient and effective experiments. (idtdna.com)
- Here we review the major developments in BAC mutagenesis in vitro . (mdpi.com)
- Once the vector is set up with flanking restriction sites, all manipulations (i.e., mutagenesis, sequencing, expression) can be performed in the same plasmid. (wikipedia.org)
- The Q5 ® Site-Directed Mutagenesis Kit enables rapid, site-specific mutagenesis of double-stranded plasmid DNA in less than 2 hours. (neb.com)
- Ultramer Oligonucleotides Mutagenesis Application Guide - Experimental Overview, Protocol, Troubleshooting" (PDF). (wikipedia.org)
- Oligonucleotide-directed Mutagenesis by Elimination of a Unique Restriction Site (USE Mutagenesis) Protocol for oligonucleotide-directed mutagenesis by elimination of a unique restriction site (USE mutagenesis). (molecularstation.com)
- Oligonucleotide-directed Mutagenesis of Single-stranded DNA Protocol Protocool for oligonucleotide-directed mutagenesis of single-stranded DNA. (molecularstation.com)
- Rapid PCR Site-Directed Mutagenesis Protocol Protocol for rapid PCR site-directed mutagenesis. (molecularstation.com)
- Site-specific Mutagenesis by Overlap Extension Protocol Protocol for site-specific mutagenesis by overlap extension. (molecularstation.com)
- See my 'Round-the-horn site-directed mutagenesis protocol to get around these limitations. (openwetware.org)
- Translesion DNA synthesis and mutagenesis in eukaryotes. (nih.gov)
- Incomplete primer synthesis can lead to errors at the mutagenesis site. (neb.com)
- Site-directed mutagenesis techniques allow specific introduction of variations at desired locations within DNA, without the need for complete de novo DNA synthesis. (thermofisher.com)
- Integrated DNA Technologies (IDT), the world leader in oligonucleotide synthesis, has recently released its free to download mutagenesis applications guide. (idtdna.com)
- Lethal mutagenesis, the killing of a microbial pathogen with a chemical mutagen, is a potential broad-spectrum antiviral treatment. (genetics.org)
- Surprisingly, models of lethal mutagenesis indicate that bacteria may be candidates for lethal mutagenesis. (genetics.org)
- and are therefore not considered candidates for lethal mutagenesis. (genetics.org)
- Lethal mutagenesis has not been proposed as a control strategy for bacteria. (genetics.org)
- Collectively, these possibilities motivate the calculation of a lethal mutagenesis threshold for bacteria, which we attempt here. (genetics.org)
- Thus, "lethal mutagenesis" driven by the class of deoxynucleoside analogs represented by 5-OH-dC could provide a new approach to treating HIV infections and, potentially, other viral infections. (pnas.org)
- Here, we examined the capacity of mutagenic deoxynucleoside analogs to modify the fidelity of replication of HIV during sequential passages in culture and to induce lethal mutagenesis. (pnas.org)
- This new edition explores current and emerging mutagenesis methods focusing specifically on mammalian systems and commonly used model organisms through comprehensive coverage and detailed protocols. (springer.com)
- Protocols and information related to mutagenesis and site directed mutagenesis. (molecularstation.com)
- In addition, optimized protocols and reagents reduced the length of the PCR and DpnI selection steps, providing up to a three-fold reduction in overall mutagenesis turn-around time. (genengnews.com)
- Unlike many site directed mutagenesis methods, cassette mutagenesis also does not involve primer extension by DNA polymerase. (wikipedia.org)
- The use of a master mix, a unique multi-enzyme KLD enzyme mix, and a fast polymerase ensures that, for most plasmids, the mutagenesis reaction is complete in less than two hours. (neb.com)
- Hutchison later produced with his collaborator Michael Smith in 1978 a more flexible approach to site-directed mutagenesis by using oligonucleotides in a primer extension method with DNA polymerase. (wikipedia.org)
- Supplying a wealth of useful details, the guide describes traditional mutagenesis approaches, including site directed mutagenesis by polymerase chain reaction (PCR), primer extension, inverse PCR and cassette-based methods. (idtdna.com)
- When studying protein functions, cassette mutagenesis can allow a scientist to change individual amino acids by introducing different codons or omitting codons. (wikipedia.org)
- High-frequency S-layer protein variation in Campylobacter fetus revealed by sapA mutagenesis. (molecularstation.com)
- Also called site-specific mutagenesis or oligonucleotide-directed mutagenesis , it is used for investigating the structure and biological activity of DNA , RNA , and protein molecules, and for protein engineering . (wikipedia.org)
- Combinatorial mutagenesis and pseudorevertant analysis to characterize regions in loop E of the CP47 protein in Synechocystis sp. (ingentaconnect.com)
- Mutagenesis experiments allow researchers to modulate protein activity and characterize structure- function relationships, which enriches our understanding of basic cellular processes and disease mechanisms and fuels discoveries in new therapies for complex diseases such as cancer. (genengnews.com)
- Ixsys is currently using the Kauffman-Ballivet technology in conjunction with its other proprietary protein engineering technologies, such as Codon-Based Mutagenesis , to optimize the function of antibodies with therapeutic potential. (thefreedictionary.com)
- The guide also covers random mutagenesis, a technique which is often used to optimize protein expression or function by creating unpredictable alterations. (idtdna.com)
- This method was effectively applied for multi-site-directed mutagenesis directly from mouse genomic DNA, as well as for combination, deletion or insertion of DNA fragments. (biomedsearch.com)
- The bookcovers genesis, linker scanning mutagenesis, nested deletion mutagenesis, and a specialized E coli mutator strain. (whsmith.co.uk)
- Identification of a locus involved in meningococcal lipopolysaccharide biosynthesis by deletion mutagenesis. (biomedsearch.com)
- A novel method for insertion/deletion mutagenesis in meningococci was devised. (biomedsearch.com)
- In contrast, NEBaseChanger , a free online primer design tool, has been developed not only to aid in the design of primers for the Q5 Site-Directed Mutagenesis Kit , but also to provide an annealing temperature for Q5 that accounts for mismatched nucleotides. (neb.com)
- For optimum annealing conditions, we propose a length of 30-35 nucleotides for the mutagenesis reverse primer. (thefreedictionary.com)
- I. European patent No. 0 752 008 with the title 'DNA mutagenesis by random fragmentation and reassembly. (epo.org)
- In this volume experts from industrial and academic laboratories describe easily reproducible and up-to-date procedures for site directed and random mutagenesis. (whsmith.co.uk)
- Here, we describe a novel technique for random transposon mutagenesis using a mariner -based transposon into C. jejuni strain 480. (asm.org)
- We have demonstrated, for the first time, random in vivo transposon mutagenesis of C. jejuni . (asm.org)
- By deleting the calcium binding loop from subtilisin, the enzyme was destabilized, and the analysis of the structure and stability of the loop-deleted prototype followed by directed mutagenesis and selection for increased stability resulted in a subtilisin mutant with native-like proteolytic activity but 1000-times greater stability in strongly chelating conditions. (thefreedictionary.com)
- If you have problems with this procedure, you can try 'Round-the-horn site-directed mutagenesis which uses PCR to amplify the desired mutant product. (openwetware.org)
- a) Overlap extension method for site‐directed mutagenesis. (els.net)
- Presently, the overlap extension method, megaprimer method, Quick Change method (Stratagene) are prevalent among numerous PCR-based mutagenesis methods developed commercially or non-commercially. (protocol-online.org)
- In this article, the traditional multi-site-directed mutagenesis method based on overlap extension PCR was improved specifically for complicated templates, such as genomic sequence or complementary DNA. (biomedsearch.com)
- Mutagenesis is largely the outcome of insults to DNA by environmental agents including alkylating agents and by endogenous metabolic oxidative metabolites such as reactive oxygen, and plays an important role in initiation, progression, and ultimately formation of cancer ( Wang, 2001 ). (intechopen.com)
- Организаторы: The Environmental Mutagenesis and Genomics Society. (science-community.org)
- All site‐directed alterations requiring site‐directed mutagenesis technique are done at the DNA level. (els.net)
- Q5® Site‐Directed Mutagenesis Kit. (els.net)
- b) SLiCE‐mediated PCR‐based site‐directed mutagenesis (SLiP site‐directed mutagenesis). (els.net)
- Figure 1: Site-specific mutagenesis proceeds in less than 2 hours. (neb.com)
- NEW YORK (GenomeWeb) - Researchers at the University of Washington have developed a method to enable researchers to perform site-directed, single amino acid mutagenesis. (genomeweb.com)
- Conventional techniques to perform site-directed mutagenesis at five sites (along with accompanying miniprep and sequencing reactions) can take up to 15 days to complete. (genengnews.com)
- I met such problem also when I was using stratagene site-directed mutagenesis kit. (protocol-online.org)
- For more information on the Gibson Assembly Site-Directed Mutagenesis Kit, please visit www. (thefreedictionary.com)
- Site directed mutagenesis of these four residues has indicated that His140 is the only histidine required for catalytic activity. (thefreedictionary.com)
- They point out that enzyme engineering through site-directed mutagenesis will provide additional avenues for refining the enzymes' specificites. (thefreedictionary.com)
- This system replaces the popular GeneTailor™ Site-Directed Mutagenesis System, and has been completely redesigned to be at the leading edge of commercial site-directed mutagenesis products currently available on the market. (thermofisher.com)
- Therefore, this receptor family serves as an excellent example for demonstrating the suitability of site-directed mutagenesis for investigating receptor function and exploring drug action. (intechopen.com)
- In order to develop a more "human-like" uricase for therapeutic use, exon replacement/restoration and site-directed mutagenesis were performed to obtain porcine-human uricase with higher homology to deduced human uricase (dHU) and increased uricolytic activity. (mdpi.com)
- Site-directed mutagenesis can be used to change particular base pairs in a piece of DNA. (openwetware.org)
- The approach described here is adapted from the Stratagene site-directed mutagenesis kit, the manual can be found here . (openwetware.org)
- 1987. Introduction of H-2Dd determinants into the H-2Ld antigen by site directed mutagenesis. (springer.com)
- Since several studies focused on key role of Asp90 in L-ASNase active site, we developed our stud to substitution of Asp90 through rational site directed mutagenesis method. (ukessays.com)