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.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Sequence Analysis, DNA: A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.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.DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.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.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.Genes, Bacterial: The functional hereditary units of BACTERIA.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).DNA Restriction Enzymes: Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.Conjugation, Genetic: A parasexual process in BACTERIA; ALGAE; FUNGI; and ciliate EUKARYOTA for achieving exchange of chromosome material during fusion of two cells. In bacteria, this is a uni-directional transfer of genetic material; in protozoa it is a bi-directional exchange. In algae and fungi, it is a form of sexual reproduction, with the union of male and female gametes.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.Nucleic Acid Hybridization: Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)Restriction Mapping: Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.R Factors: A class of plasmids that transfer antibiotic resistance from one bacterium to another by conjugation.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.Bacterial Proteins: Proteins found in any species of bacterium.Repetitive Sequences, Nucleic Acid: Sequences of DNA or RNA that occur in multiple copies. There are several types: INTERSPERSED REPETITIVE SEQUENCES are copies of transposable elements (DNA TRANSPOSABLE ELEMENTS or RETROELEMENTS) dispersed throughout the genome. TERMINAL REPEAT SEQUENCES flank both ends of another sequence, for example, the long terminal repeats (LTRs) on RETROVIRUSES. Variations may be direct repeats, those occurring in the same direction, or inverted repeats, those opposite to each other in direction. TANDEM REPEAT SEQUENCES are copies which lie adjacent to each other, direct or inverted (INVERTED REPEAT SEQUENCES).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.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.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.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.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.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.DNA Replication: The process by which a DNA molecule is duplicated.DNA, Viral: Deoxyribonucleic acid that makes up the genetic material of viruses.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.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.Phylogeny: The relationships of groups of organisms as reflected by their genetic makeup.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.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.Chromosome Mapping: Any method used for determining the location of and relative distances between genes on a chromosome.Chromosomes, Bacterial: Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.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.Nucleic Acid Conformation: The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.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, Fungal: Deoxyribonucleic acid that makes up the genetic material of fungi.Extrachromosomal Inheritance: Vertical transmission of hereditary characters by DNA from cytoplasmic organelles such as MITOCHONDRIA; CHLOROPLASTS; and PLASTIDS, or from PLASMIDS or viral episomal DNA.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.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.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.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).F Factor: A plasmid whose presence in the cell, either extrachromosomal or integrated into the BACTERIAL CHROMOSOME, determines the "sex" of the bacterium, host chromosome mobilization, transfer via conjugation (CONJUGATION, GENETIC) of genetic material, and the formation of SEX PILI.Replicon: Any DNA sequence capable of independent replication or a molecule that possesses a REPLICATION ORIGIN and which is therefore potentially capable of being replicated in a suitable cell. (Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)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.DNA, Circular: Any of the covalently closed DNA molecules found in bacteria, many viruses, mitochondria, plastids, and plasmids. Small, polydisperse circular DNA's have also been observed in a number of eukaryotic organisms and are suggested to have homology with chromosomal DNA and the capacity to be inserted into, and excised from, chromosomal DNA. It is a fragment of DNA formed by a process of looping out and deletion, containing a constant region of the mu heavy chain and the 3'-part of the mu switch region. Circular DNA is a normal product of rearrangement among gene segments encoding the variable regions of immunoglobulin light and heavy chains, as well as the T-cell receptor. (Riger et al., Glossary of Genetics, 5th ed & Segen, Dictionary of Modern Medicine, 1992)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.Electrophoresis, Agar Gel: Electrophoresis in which agar or agarose gel is used as the diffusion medium.Bacteriocin Plasmids: Plasmids encoding bacterial exotoxins (BACTERIOCINS).Drug Resistance, Microbial: The ability of microorganisms, especially bacteria, to resist or to become tolerant to chemotherapeutic agents, antimicrobial agents, or antibiotics. This resistance may be acquired through gene mutation or foreign DNA in transmissible plasmids (R FACTORS).Genes, Regulator: Genes which regulate or circumscribe the activity of other genes; specifically, genes which code for PROTEINS or RNAs which have GENE EXPRESSION REGULATION functions.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.Gene Expression Regulation, Bacterial: Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria.Escherichia coli Proteins: Proteins obtained from ESCHERICHIA COLI.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.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.Genetic Variation: Genotypic differences observed among individuals in a population.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.DNA, Superhelical: Circular duplex DNA isolated from viruses, bacteria and mitochondria in supercoiled or supertwisted form. This superhelical DNA is endowed with free energy. During transcription, the magnitude of RNA initiation is proportional to the DNA superhelicity.Base Composition: The relative amounts of the PURINES and PYRIMIDINES in a nucleic acid.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Replication Origin: A unique DNA sequence of a replicon at which DNA REPLICATION is initiated and proceeds bidirectionally or unidirectionally. It contains the sites where the first separation of the complementary strands occurs, a primer RNA is synthesized, and the switch from primer RNA to DNA synthesis takes place. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)DNA, Mitochondrial: Double-stranded DNA of MITOCHONDRIA. In eukaryotes, the mitochondrial GENOME is circular and codes for ribosomal RNAs, transfer RNAs, and about 10 proteins.Transcription Factors: Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.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.Rhizobium: A genus of gram-negative, aerobic, rod-shaped bacteria that activate PLANT ROOT NODULATION in leguminous plants. Members of this genus are nitrogen-fixing and common soil inhabitants.Evolution, Molecular: The process of cumulative change at the level of DNA; RNA; and PROTEINS, over successive generations.Regulatory Sequences, Nucleic Acid: Nucleic acid sequences involved in regulating the expression of genes.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.DNA Probes: Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA 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 DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections.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.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.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.Chromosome Deletion: Actual loss of portion of a chromosome.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).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.Bacteriophage lambda: A temperate inducible phage and type species of the genus lambda-like viruses, in the family SIPHOVIRIDAE. Its natural host is E. coli K12. Its VIRION contains linear double-stranded DNA with single-stranded 12-base 5' sticky ends. The DNA circularizes on infection.Vaccines, DNA: Recombinant DNA vectors encoding antigens administered for the prevention or treatment of disease. The host cells take up the DNA, express the antigen, and present it to the immune system in a manner similar to that which would occur during natural infection. This induces humoral and cellular immune responses against the encoded antigens. The vector is called naked DNA because there is no need for complex formulations or delivery agents; the plasmid is injected in saline or other buffers.Electroporation: A technique in which electric pulses of intensity in kilovolts per centimeter and of microsecond-to-millisecond duration cause a temporary loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA.DNA, Ribosomal: DNA sequences encoding RIBOSOMAL RNA and the segments of DNA separating the individual ribosomal RNA genes, referred to as RIBOSOMAL SPACER DNA.Genes, Fungal: The functional hereditary units of FUNGI.Molecular Weight: The sum of the weight of all the atoms in a molecule.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.Deoxyribonuclease EcoRI: One of the Type II site-specific deoxyribonucleases (EC 18.104.22.168). It recognizes and cleaves the sequence G/AATTC at the slash. EcoRI is from E coliRY13. Several isoschizomers have been identified. EC 3.1.21.-.Phenotype: The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.Biological Evolution: The process of cumulative change over successive generations through which organisms acquire their distinguishing morphological and physiological characteristics.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)Pseudomonas: A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in nature. Some species are pathogenic for humans, animals, and plants.Gene Expression: The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.Mutagenesis: Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.DNA, Satellite: Highly repetitive DNA sequences found in HETEROCHROMATIN, mainly near centromeres. They are composed of simple sequences (very short) (see MINISATELLITE REPEATS) repeated in tandem many times to form large blocks of sequence. Additionally, following the accumulation of mutations, these blocks of repeats have been repeated in tandem themselves. The degree of repetition is on the order of 1000 to 10 million at each locus. Loci are few, usually one or two per chromosome. They were called satellites since in density gradients, they often sediment as distinct, satellite bands separate from the bulk of genomic DNA owing to a distinct BASE COMPOSITION.Genes, Viral: The functional hereditary units of VIRUSES.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)Gene Amplification: A selective increase in the number of copies of a gene coding for a specific protein without a proportional increase in other genes. It occurs naturally via the excision of a copy of the repeating sequence from the chromosome and its extrachromosomal replication in a plasmid, or via the production of an RNA transcript of the entire repeating sequence of ribosomal RNA followed by the reverse transcription of the molecule to produce an additional copy of the original DNA sequence. Laboratory techniques have been introduced for inducing disproportional replication by unequal crossing over, uptake of DNA from lysed cells, or generation of extrachromosomal sequences from rolling circle replication.Tetracycline: A naphthacene antibiotic that inhibits AMINO ACYL TRNA binding during protein synthesis.Gene Transfer, Horizontal: The naturally occurring transmission of genetic information between organisms, related or unrelated, circumventing parent-to-offspring transmission. Horizontal gene transfer may occur via a variety of naturally occurring processes such as GENETIC CONJUGATION; GENETIC TRANSDUCTION; and TRANSFECTION. It may result in a change of the recipient organism's genetic composition (TRANSFORMATION, GENETIC).Gene Transfer Techniques: The introduction of functional (usually cloned) GENES into cells. A variety of techniques and naturally occurring processes are used for the gene transfer such as cell hybridization, LIPOSOMES or microcell-mediated gene transfer, ELECTROPORATION, chromosome-mediated gene transfer, TRANSFECTION, and GENETIC TRANSDUCTION. Gene transfer may result in genetically transformed cells and individual organisms.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.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.Software: Sequential operating programs and data which instruct the functioning of a digital computer.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.Bacillus subtilis: A species of gram-positive bacteria that is a common soil and water saprophyte.DNA, Plant: Deoxyribonucleic acid that makes up the genetic material of plants.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.Recombinant Proteins: Proteins prepared by recombinant DNA technology.RNA, Bacterial: Ribonucleic acid in bacteria having regulatory and catalytic roles as well as involvement in protein synthesis.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.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.DNA Mutational Analysis: Biochemical identification of mutational changes in a nucleotide sequence.Single-Strand Specific DNA and RNA Endonucleases: Enzymes that catalyze the endonucleolytic cleavage of single-stranded regions of DNA or RNA molecules while leaving the double-stranded regions intact. They are particularly useful in the laboratory for producing "blunt-ended" DNA molecules from DNA with single-stranded ends and for sensitive GENETIC TECHNIQUES such as NUCLEASE PROTECTION ASSAYS that involve the detection of single-stranded DNA and RNA.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.Bacteriophages: Viruses whose hosts are bacterial cells.Kinetics: The rate dynamics in chemical or physical systems.Nucleic Acid Renaturation: The reformation of all, or part of, the native conformation of a nucleic acid molecule after the molecule has undergone denaturation.Sequence Deletion: Deletion of sequences of nucleic acids from the genetic material of an individual.Chromosomes: In a prokaryotic cell or in the nucleus of a eukaryotic cell, a structure consisting of or containing DNA which carries the genetic information essential to the cell. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)Plant Tumors: A localized proliferation of plant tissue forming a swelling or outgrowth, commonly with a characteristic shape and unlike any organ of the normal plant. Plant tumors or galls usually form in response to the action of a pathogen or a pest. (Holliday, P., A Dictionary of Plant Pathology, 1989, p330)Deoxyribonuclease I: An enzyme capable of hydrolyzing highly polymerized DNA by splitting phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide. This catalyzes endonucleolytic cleavage of DNA yielding 5'-phosphodi- and oligonucleotide end-products. The enzyme has a preference for double-stranded DNA.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.Salmonella typhimurium: A serotype of Salmonella enterica that is a frequent agent of Salmonella gastroenteritis in humans. It also causes PARATYPHOID FEVER.DNA, Single-Stranded: A single chain of deoxyribonucleotides that occurs in some bacteria and viruses. It usually exists as a covalently closed circle.Genome, Bacterial: The genetic complement of a BACTERIA as represented in its DNA.Chloramphenicol O-Acetyltransferase: An enzyme that catalyzes the acetylation of chloramphenicol to yield chloramphenicol 3-acetate. Since chloramphenicol 3-acetate does not bind to bacterial ribosomes and is not an inhibitor of peptidyltransferase, the enzyme is responsible for the naturally occurring chloramphenicol resistance in bacteria. The enzyme, for which variants are known, is found in both gram-negative and gram-positive bacteria. EC 22.214.171.124.Kanamycin: Antibiotic complex produced by Streptomyces kanamyceticus from Japanese soil. Comprises 3 components: kanamycin A, the major component, and kanamycins B and C, the minor components.Anti-Bacterial Agents: Substances that reduce the growth or reproduction of BACTERIA.Simian virus 40: A species of POLYOMAVIRUS originally isolated from Rhesus monkey kidney tissue. It produces malignancy in human and newborn hamster kidney cell cultures.Bacterial Outer Membrane Proteins: Proteins isolated from the outer membrane of Gram-negative bacteria.Deoxyribonuclease HindIII: One of the Type II site-specific deoxyribonucleases (EC 126.96.36.199). It recognizes and cleaves the sequence A/AGCTT at the slash. HindIII is from Haemophilus influenzae R(d). Numerous isoschizomers have been identified. EC 3.1.21.-.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.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.beta-Lactamases: Enzymes found in many bacteria which catalyze the hydrolysis of the amide bond in the beta-lactam ring. Well known antibiotics destroyed by these enzymes are penicillins and cephalosporins.Cosmids: Plasmids containing at least one cos (cohesive-end site) of PHAGE LAMBDA. They are used as cloning vehicles.Agrobacterium tumefaciens: A species of gram-negative, aerobic bacteria isolated from soil and the stems, leafs, and roots of plants. Some biotypes are pathogenic and cause the formation of PLANT TUMORS in a wide variety of higher plants. The species is a major research tool in biotechnology.Polymorphism, Restriction Fragment Length: Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment.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.Coliphages: Viruses whose host is Escherichia coli.Centromere: The clear constricted portion of the chromosome at which the chromatids are joined and by which the chromosome is attached to the spindle during cell division.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.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.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.Genotype: The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS.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.Enterococcus faecalis: A species of gram-positive, coccoid bacteria commonly isolated from clinical specimens and the human intestinal tract. Most strains are nonhemolytic.DNA, Ribosomal Spacer: The intergenic DNA segments that are between the ribosomal RNA genes (internal transcribed spacers) and between the tandemly repeated units of rDNA (external transcribed spacers and nontranscribed spacers).Genetic Therapy: Techniques and strategies which include the use of coding sequences and other conventional or radical means to transform or modify cells for the purpose of treating or reversing disease conditions.Trans-Activators: Diffusible gene products that act on homologous or heterologous molecules of viral or cellular DNA to regulate the expression of proteins.Algorithms: A procedure consisting of a sequence of algebraic formulas and/or logical steps to calculate or determine a given task.Deoxyribonucleases, Type II Site-Specific: Enzyme systems containing a single subunit and requiring only magnesium for endonucleolytic activity. The corresponding modification methylases are separate enzymes. The systems recognize specific short DNA sequences and cleave either within, or at a short specific distance from, the recognition sequence to give specific double-stranded fragments with terminal 5'-phosphates. Enzymes from different microorganisms with the same specificity are called isoschizomers. EC 188.8.131.52.Electrophoresis, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.Endonucleases: Enzymes that catalyze the hydrolysis of the internal bonds and thereby the formation of polynucleotides or oligonucleotides from ribo- or deoxyribonucleotide chains. EC 3.1.-.Colicins: Bacteriocins elaborated by strains of Escherichia coli and related species. They are proteins or protein-lipopolysaccharide complexes lethal to other strains of the same species.Lactococcus lactis: A non-pathogenic species of LACTOCOCCUS found in DAIRY PRODUCTS and responsible for the souring of MILK and the production of LACTIC ACID.Gene Order: The sequential location of genes on a chromosome.Nucleosomes: The repeating structural units of chromatin, each consisting of approximately 200 base pairs of DNA wound around a protein core. This core is composed of the histones H2A, H2B, H3, and H4.Genome: The genetic complement of an organism, including all of its GENES, as represented in its DNA, or in some cases, its RNA.RNA, Ribosomal: The most abundant form of RNA. Together with proteins, it forms the ribosomes, playing a structural role and also a role in ribosomal binding of mRNA and tRNAs. Individual chains are conventionally designated by their sedimentation coefficients. In eukaryotes, four large chains exist, synthesized in the nucleolus and constituting about 50% of the ribosome. (Dorland, 28th ed)Cell Nucleus: Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)Bacteriocins: Substances elaborated by specific strains of bacteria that are lethal against other strains of the same or related species. They are protein or lipopolysaccharide-protein complexes used in taxonomy studies of bacteria.Plants: Multicellular, eukaryotic life forms of kingdom Plantae (sensu lato), comprising the VIRIDIPLANTAE; RHODOPHYTA; and GLAUCOPHYTA; all of which acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations.Enhancer Elements, Genetic: Cis-acting DNA sequences which can increase transcription of genes. Enhancers can usually function in either orientation and at various distances from a promoter.Chickens: Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.Deoxyribonuclease BamHI: One of the Type II site-specific deoxyribonucleases (EC 184.108.40.206). It recognizes and cleaves the sequence G/GATCC at the slash. BamHI is from Bacillus amyloliquefaciens N. Numerous isoschizomers have been identified. EC 3.1.21.-.Tetracycline Resistance: Nonsusceptibility of bacteria to the action of TETRACYCLINE which inhibits aminoacyl-tRNA binding to the 30S ribosomal subunit during protein synthesis.DNA Nucleotidyltransferases: Enzymes that catalyze the incorporation of deoxyribonucleotides into a chain of DNA. EC 2.7.7.-.RNA: A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)Sequence Homology: The degree of similarity between sequences. Studies of AMINO ACID SEQUENCE HOMOLOGY and NUCLEIC ACID SEQUENCE HOMOLOGY provide useful information about the genetic relatedness of genes, gene products, and species.Pseudomonas putida: A species of gram-negative, aerobic bacteria isolated from soil and water as well as clinical specimens. Occasionally it is an opportunistic pathogen.Polymorphism, Genetic: The regular and simultaneous occurrence in a single interbreeding population of two or more discontinuous genotypes. The concept includes differences in genotypes ranging in size from a single nucleotide site (POLYMORPHISM, SINGLE NUCLEOTIDE) to large nucleotide sequences visible at a chromosomal level.Introns: Sequences of DNA in the genes that are located between the EXONS. They are transcribed along with the exons but are removed from the primary gene transcript by RNA SPLICING to leave mature RNA. Some introns code for separate genes.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.Computational Biology: A field of biology concerned with the development of techniques for the collection and manipulation of biological data, and the use of such data to make biological discoveries or predictions. This field encompasses all computational methods and theories for solving biological problems including manipulation of models and datasets.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.Transduction, Genetic: The transfer of bacterial DNA by phages from an infected bacterium to another bacterium. This also refers to the transfer of genes into eukaryotic cells by viruses. This naturally occurring process is routinely employed as a GENE TRANSFER TECHNIQUE.Shigella flexneri: A bacterium which is one of the etiologic agents of bacillary dysentery (DYSENTERY, BACILLARY) and sometimes of infantile gastroenteritis.DNA, Chloroplast: Deoxyribonucleic acid that makes up the genetic material of CHLOROPLASTS.Klebsiella pneumoniae: Gram-negative, non-motile, capsulated, gas-producing rods found widely in nature and associated with urinary and respiratory infections in humans.Genetic Markers: A phenotypically recognizable genetic trait which can be used to identify a genetic locus, a linkage group, or a recombination event.Nucleic Acid Denaturation: Disruption of the secondary structure of nucleic acids by heat, extreme pH or chemical treatment. Double strand DNA is "melted" by dissociation of the non-covalent hydrogen bonds and hydrophobic interactions. Denatured DNA appears to be a single-stranded flexible structure. The effects of denaturation on RNA are similar though less pronounced and largely reversible.Substrate Specificity: A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.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.Kanamycin Resistance: Nonsusceptibility of bacteria to the antibiotic KANAMYCIN, which can bind to their 70S ribosomes and cause misreading of messenger RNA.DNA Footprinting: A method for determining the sequence specificity of DNA-binding proteins. DNA footprinting utilizes a DNA damaging agent (either a chemical reagent or a nuclease) which cleaves DNA at every base pair. DNA cleavage is inhibited where the ligand binds to DNA. (from Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)Globins: A superfamily of proteins containing the globin fold which is composed of 6-8 alpha helices arranged in a characterstic HEME enclosing structure.Chromatin: The material of CHROMOSOMES. It is a complex of DNA; HISTONES; and nonhistone proteins (CHROMOSOMAL PROTEINS, NON-HISTONE) found within the nucleus of a cell.DNA-Directed RNA Polymerases: Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992).RNA, Ribosomal, 16S: Constituent of 30S subunit prokaryotic ribosomes containing 1600 nucleotides and 21 proteins. 16S rRNA is involved in initiation of polypeptide synthesis.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.Nucleotide Mapping: Two-dimensional separation and analysis of nucleotides.Luciferases: Enzymes that oxidize certain LUMINESCENT AGENTS to emit light (PHYSICAL LUMINESCENCE). The luciferases from different organisms have evolved differently so have different structures and substrates.Genetic Code: The meaning ascribed to the BASE SEQUENCE with respect to how it is translated into AMINO ACID SEQUENCE. The start, stop, and order of amino acids of a protein is specified by consecutive triplets of nucleotides called codons (CODON).Penicillinase: A beta-lactamase preferentially cleaving penicillins. (Dorland, 28th ed) EC 3.5.2.-.Reading Frames: The three possible sequences of CODONS by which GENETIC TRANSLATION may occur from one nucleotide sequence. A segment of mRNA 5'AUCCGA3' could be translated as 5'AUC.. or 5'UCC.. or 5'CCG.., depending on the location of the START CODON.Bacteriophage P1: A species of temperate bacteriophage in the genus P1-like viruses, family MYOVIRIDAE, which infects E. coli. It is the largest of the COLIPHAGES and consists of double-stranded DNA, terminally redundant, and circularly permuted.Lysogeny: The phenomenon by which a temperate phage incorporates itself into the DNA of a bacterial host, establishing a kind of symbiotic relation between PROPHAGE and bacterium which results in the perpetuation of the prophage in all the descendants of the bacterium. Upon induction (VIRUS ACTIVATION) by various agents, such as ultraviolet radiation, the phage is released, which then becomes virulent and lyses the bacterium.Genomics: The systematic study of the complete DNA sequences (GENOME) of organisms.Exons: The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA.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.DNA, Intergenic: Any of the DNA in between gene-coding DNA, including untranslated regions, 5' and 3' flanking regions, INTRONS, non-functional pseudogenes, and non-functional repetitive sequences. This DNA may or may not encode regulatory functions.Fungal Proteins: Proteins found in any species of fungus.Gene Dosage: The number of copies of a given gene present in the cell of an organism. An increase in gene dosage (by GENE DUPLICATION for example) can result in higher levels of gene product formation. GENE DOSAGE COMPENSATION mechanisms result in adjustments to the level GENE EXPRESSION when there are changes or differences in gene dosage.Polyomavirus: A genus of potentially oncogenic viruses of the family POLYOMAVIRIDAE. These viruses are normally present in their natural hosts as latent infections. The virus is oncogenic in hosts different from the species of origin.
Freeware DNA cloning, sequence analysis and plasmid/DNA plotting software. *. Čermák V. "Restriction Analyzer". Retrieved 2016- ... and the position of their DNA cleavage site relative to the target sequence. DNA sequence analyses of restriction ... Recognition sequences in DNA differ for each restriction enzyme, producing differences in the length, sequence and strand ... but the forward and backward sequences are found in complementary DNA strands (i.e., of double-stranded DNA), as in GTATAC ( ...
i have aslo included the DNA sequence of the plasmid for this article. i have also included two references that can be verified ... The speedy deletion might be due to the large DNA sequence data i have added. please delete the notice if no one has any ... I see nothing wrong with having the sequence. What the article needs is, first, full citations in the usual academic style to ...
... is a database that can be used to remove vector contamination from DNA sequences. Plasmid Cochrane, Guy R; Galperin ...
A cosmid is a type of hybrid plasmid that contains a Lambda phage cos sequence. Cosmids (cos sites + plasmid = cosmids) DNA ... Cos sequences are ~200 base pairs long and essential for packaging. They contain a cosN site where DNA is nicked at each strand ... Plasmids become unstable after a certain amount of DNA has been inserted into them, because their increased size is more ... Target DNA: the genomic DNA to be cloned has to be cut into the appropriate size range of restriction fragments. This is ...
Freeware DNA cloning, sequence analysis and plasmid/DNA plotting software Čermák V. "Restriction Analyzer". Retrieved 2016-08- ... the nature of their target sequence, and the position of their DNA cleavage site relative to the target sequence. DNA sequence ... All types of enzymes recognize specific short DNA sequences and carry out the endonucleolytic cleavage of DNA to give specific ... Recognition sequences in DNA differ for each restriction enzyme, producing differences in the length, sequence and strand ...
DNA sequence identity. Chlamydiaceae species have varying inclusion morphology, varying extrachromosomal plasmid content, and ...
Then, the DNA sequence can be inserted back to a random location of the genome. DNA transposons are a DNA segment that can move ... Plasmids of bacteria are a transferable genetic element through bacterial conjugation. This is a mechanism of horizontal gene ... are DNA sequences that can move locations within a genome, which includes retrotransposons and DNA transposons. ... MGEs such as plasmids by a horizontal transmission are generally beneficial to an organism. The ability of transferring ...
DNA sequence and mechanism of transfer". Journal of Bacteriology. 182 (1): 81-90. doi:10.1128/jb.182.1.81-90.2000. PMC 94243 . ... He has found two circular chromosomes or plasmids in Rhodobacter with Kaplan S in 1989. He received an award as cum laude from ... Kwong, S. M.; Yeo, C. C.; Suwanto, A; Poh, C. L. (2000). "Characterization of the endogenous plasmid from Pseudomonas ... Seumahu, C. A.; Suwanto, A.; Rusmana, I.; Solihin, D. D. (2012). "Comparison of DNA Extraction Methods for Microbial Community ...
Ori is the DNA sequence that signals for the origin of replication, sometimes referred to simply as origin. In E. coli, ori is ... The plasmid ori sequences are similar to oriC, and are called oriV (origin of vegetative replication). During conjugation, the ... Mitochondrial DNA in many organisms has two ori sequences. In humans, they are called oriH and oriL for the heavy and light ... sequence of the FAT plasmid. Bacteria have a single origin for replication. Eukaryotes have multiple replicons, each with an ...
The idea arose that different DNA sequences could be inserted into a plasmid and that these foreign sequences would be carried ... The desired products (vector DNA covalently linked to foreign DNA) will be present, but other sequences (e.g. foreign DNA ... vector DNA with no recombinant sequence inserted). In these vectors, foreign DNA is inserted into a sequence that encodes an ... termed recombinant DNA. By recombining DNA segments of interest with vector DNA, such as bacteriophage or plasmids, which ...
Plasmid DNA sequences can be manufactured using the hTERT promoter followed by genes encoding for specific proteins. The ... This addition of repetitive DNA sequences prevents degradation of the chromosomal ends following multiple rounds of replication ... Jan HM, Wei MF, Peng CL, Lin SJ, Lai PS, Shieh MJ (January 2012). "The use of polyethylenimine-DNA to topically deliver hTERT ... were seen in vitro when combining the use of antisense hTERT sequences with the introduction of a tumor-suppressing plasmid by ...
The plasmid is transformed into bacteria and the identity of the insert is confirmed by DNA sequencing. Cloning Molecular ... After a good number of bacterial colonies have grown, they can be miniprepped to harvest the plasmid DNA. In order to ensure ... In molecular biology, subcloning is a technique used to move a particular DNA sequence from a parent vector to a destination ... The plasmid is often transformed into a bacterium like E. coli. Ideally when the bacterium divides the plasmid should also be ...
DNA sequence of the region from 89.2 to 92.8 minutes. Nucleic Acids Res. 1993 Nov 25;21(23):5408-17.y Brosius J. Plasmid ... Some of his scientific contributions involve the first genetic sequencing of a ribosomal RNA operon, the design of plasmids for ... There, he sequenced the first large ribosomal RNAs via their genes utilizing the Maxam-Gilbert sequencing method. It took ~2.5 ... Protein sequence analysis: automated microsequencing. Science. 1983 Feb 11;219(4585):650-9. Brosius J, Dull TJ, Sleeter DD, ...
"DNA sequence and comparison of virulence plasmids from Rhodococcus equi ATCC 33701 and 103". Infection and Immunity. 68 (12): ... These circular plasmids consist of a conserved backbone responsible for replication and bacterial conjugation of the plasmid. ... All strains isolated from foals and the majority of human, cattle, and pig isolates contain a large plasmid. This plasmid has ... This portion of the plasmid is highly conserved and found in nonpathogenic Rhodococci plasmids. In addition to the conserved ...
Every BioBrick part is a DNA sequence which is carried by a circular plasmid, which acts a vector. The vector acts as a ... T5 exonuclease attacks the 5' ends of sequences, creating single-stranded DNA in the ends of all sequences where the different ... which digests methylated DNA like plasmids. Eliminating the template plasmids with DpnI leaves only the insert to be amplified ... ends of the DNA part respectively. These standard sequences encode specific restriction enzyme sites. The prefix sequence ...
Restriction endonucleases and DNA ligase are then used to clone the sequences into bacterial plasmids. The cloned bacteria are ... This sscDNA is converted into a double stranded DNA with the help of DNA polymerase. However, for DNA polymerase to synthesize ... cDNA library lacks the non-coding and regulatory elements found in genomic DNA. Genomic DNA libraries provide more detailed ... Both the cDNA and the linker have blunt ends which can be ligated together using a high concentration of T4 DNA ligase. Then ...
For decoy DNA delivery. Decoy DNA is an exogenous double-strand DNA (dsDNA), which can mimic a promoter sequence that can ... For plasmid delivery. Individual genes can be inserted into specific sites on plasmids, and recombinant plasmids can be ... A method using macro-branched TAT has been proposed for plasmid DNA delivery into various cell lines and showed significant ... Nucleic acid-based macromolecules such as siRNA, antisense oligonucleotide, decoy DNA, and plasmid have been realized as ...
Cosmid vectors are plasmids that contain a small region of bacteriophage λ DNA called the cos sequence. This sequence allows ... A plasmid is a double stranded circular DNA molecule commonly used for molecular cloning. Plasmids are generally 2 to 4 ... for use in DNA sequencing. The importance of this success contributed to the ever-increasing demand for sequencing genomes to ... which are more manageable for sequencing. Once a clone from a genomic library is sequenced, the sequence can be used to screen ...
repE for plasmid replication and regulation of copy number. parA and parB for partitioning F plasmid DNA to daughter cells ... A short piece of the organism's DNA is amplified as an insert in BACs, and then sequenced. Finally, the sequenced parts are ... A bacterial artificial chromosome (BAC) is a DNA construct, based on a functional fertility plasmid (or F-plasmid), used for ... A similar cloning vector called a PAC has also been produced from the DNA of P1 bacteriophage. BACs are often used to sequence ...
The linear DNA genome of Lautropia mirabilis type strain ATCC 51599 is completely sequenced. There is only one chromosome which ... has a length of approximately 3.15 Mb , and no plasmids have been discovered . This genome size is small compared to other ...
OriC (Origin of Replication): The sequence starting with which the plasmid-DNA will be replicated in the recipient cell. tra- ... The F plasmid contains only F factor DNA and no DNA from the bacterial genome. F' (F-prime) bacteria are formed by incorrect ... The first DNA helicase ever described is encoded on the F-plasmid and is responsible for initiating plasmid transfer. It was ... Unlike other plasmids, F factor is constitutive for transfer proteins due to the gene traJ. The F plasmid belongs to a class of ...
DNA sequences to create recombinant DNA. Plasmids, discovered in 1952, became important tools for transferring information ... The T-DNA region of this plasmid is responsible for insertion of the DNA. The DNA to be inserted is cloned into a binary vector ... the plasmid construct must also contain T-DNA. Agrobacterium naturally inserts DNA from a tumor-inducing plasmid into any ... If the DNA sequence of the gene and the organism is known, restriction enzymes that will cut the DNA either side of the gene ...
The DNA sequence of the VRSA vanA gene was identical to that of a vancomycin-resistant strain of Enterococcus faecalis ... The vanA gene was later found to be encoded within a transposon located on a plasmid carried by the VRSA isolate. This ...
... (PAM) is a 2-6 base pair DNA sequence immediately following the DNA sequence targeted by the Cas9 ... But the protospacer in the invading virus or plasmid will contain the PAM sequence, and will thus be cleaved by the Cas9 ... Cas9 will not successfully bind to or cleave the target DNA sequence if it is not followed by the PAM sequence. PAM is an ... crRNA complex in recognizing gene sequences having a PAM sequence at the 3'-end. The canonical PAM is the sequence 5'-NGG-3' ...
... s are directly repeated DNA sequences which play an important role in regulation of plasmid copy number in bacterial ... This operator contains half of the iteron sequence making it able to bind the dimer and promote gene expression. Plasmids ... If a plasmid contains an extra supply of iterons outside of the saturation site it has been shown this can decrease plasmid ... Dhruba K. Chattoraj (2000). "Control of plasmid DNA replication by iterons: no longer paradoxical". Molecular Biology. 37 (3): ...
... and first sequenced in 1986-when two Japanese research teams sequenced the chloroplast DNA of liverwort and tobacco. Since ... but blank plasmids, with no coding DNA, have also been found. ... Over time, base changes in the DNA sequence can arise from ... DNA replicationEdit. Leading model of cpDNA replicationEdit. Chloroplast DNA replication via multiple D loop mechanisms. ... Chloroplast DNA Interactive gene map of chloroplast DNA from Nicotiana tabacum. Segments with labels on the inside reside on ...
Escherichia coli B171 plasmid pB171 DNA, complete sequence Escherichia coli B171 plasmid pB171 DNA, complete sequence. gi, ... Escherichia coli B171 plasmid pB171 DNA, complete sequence. NCBI Reference Sequence: NC_002142.1 ... Find in this Sequence Finds sub-sequences or patterns in the sequence and highlights the matching regions. The tool works with ... Finds sub-sequence or patterns in the sequence and highlights the matching region. The tool works with standard single letter ...
PCC 7120 plasmid pCC7120beta DNA, complete sequence Nostoc sp. PCC 7120 plasmid pCC7120beta DNA, complete sequence. gi,17158637 ... PCC 7120 plasmid pCC7120beta DNA, complete sequence. NCBI Reference Sequence: NC_003240.1 ... Find in this Sequence Finds sub-sequences or patterns in the sequence and highlights the matching regions. The tool works with ... Finds sub-sequence or patterns in the sequence and highlights the matching region. The tool works with standard single letter ...
High-frequency germ-line transmission of plasmid DNA sequences injected into fertilized zebrafish eggs. P Culp, C Nüsslein- ... High-frequency germ-line transmission of plasmid DNA sequences injected into fertilized zebrafish eggs ... High-frequency germ-line transmission of plasmid DNA sequences injected into fertilized zebrafish eggs ... High-frequency germ-line transmission of plasmid DNA sequences injected into fertilized zebrafish eggs ...
Plasmid DNA Sequencing with Universal Primers - (Mar/08/2007 ). Hi all, Ive been having some trouble with sequencing. The gene ... giving bad sequence reads. I use 1ul for plasmid DNA eg read lenghts of 900bp+. 0.5ul for PCR prdocuts.. read lenghts around ... Also use of sequencing buffer is a must for reliable results. The next important thing the amount of DNA and Big Dye terminator ... the problem is probably due to the purity of your plasmid. The DNA has to be the cleanest you can possibly get it to be. No RNA ...
... plasmids, or cDNA in as little as 90 minutes, with a low DNA input requirement. ... Prepare sequencing libraries for small genomes, PCR amplicons, ... 16s rRNA Sequencing, Amplicon Sequencing , De Novo Sequencing ... Shotgun Sequencing , Whole-Genome Sequencing. Amplicon Sequencing , De Novo Sequencing , Shotgun Sequencing , Whole-Genome ... Prepare sequencing libraries for small genomes, PCR amplicons, plasmids, or cDNA in as little as 90 minutes, with a low DNA ...
... DNA Sequence of a ColV Plasmid and Prevalence of Selected Plasmid-Encoded Virulence Genes among Avian Escherichia coli Strains ... DNA Sequence of a ColV Plasmid and Prevalence of Selected Plasmid-Encoded Virulence Genes among Avian Escherichia coli Strains ...
Bacillus anthracis can be identified on the basis of the detection of virulence factor genes located on two plasmids, pXO1 and ... and characterization of Bacillus anthracis by multiplex PCR analysis of sequences on plasmids pXO1 and pXO2 and chromosomal DNA ... Bacillus anthracis can be identified on the basis of the detection of virulence factor genes located on two plasmids, pXO1 and ... and simultaneous confirmation of the species identity independent of plasmid content. The assay amplifies lef, cya, pag (pXO1) ...
The DNA sequence in the region of the fusion between bla and tRNAArg5 genes is displayed above the plasmid map. The tRNAArg5 ... The DNA sequence in the region of the fusion between bla and tRNAArg5 genes is displayed above the plasmid map. The tRNAArg5 ... The DNA sequence in the region of the fusion between bla and tRNAArg5 genes is displayed above the plasmid map. The tRNAArg5 ... To test this we used plasmid constructs that were identical, except that the UTR DNA sequence was the wild‐type (pRL11), LV‐1 ...
The presence of pXO2 ORF sequences in other Bacillus species suggests the possibility that certain pXO2 plasmid gene functions ... Dot-blot DNA hybridizations between pXO2 ORF fragments and total genomic DNA from AWO6 were consistent with the PCR assay ... Sequences similar to the B. anthracis cap genes or their regulator, acpA, were not detected among any of the examined isolates ... The presence of pXO2 sequences in the other Bacillus isolates did not correlate with genomic relatedness established by AFLP ...
DNA sequencing. Libraries were prepared from nebulized, size-fractionated plasmid DNA (63) in the M13 Janus vector (24). DNA ... enterocolitica plasmids differ from those of LCR plasmids from the other two yersiniae (12, 32, 68, 83). DNA sequencing has ... DNA Sequencing and Analysis of the Low-Ca2+-Response Plasmid pCD1 of Yersinia pestis KIM5. Robert D. Perry,1,* Susan C. Straley ... we have sequenced one entire LCR plasmid. We chose to sequence the LCR plasmid pCD1 of Y. pestis KIM5 (for Kurdistan Iran man) ...
DNA sequencing of a plasmid with virulence from marine fish pathogen Vibrio anguillarum ... The whole length of obtained pEIB1 DNA sequence was 66 164 bp, and the overall G+C content of DNA sequence is 42.7%. This ... Virulence plasmid pJM1 prevents the conjugal entry of plasmid DNA into the marine fish pathogen Vibrio anguillarum 775. Journal ... DNA sequencing of a plasmid with virulence from marine fish pathogen Vibrio anguillarum. Wu, H-Zhen.; Zhang, H-Zhan.; Lü, C-Xia ...
Genomic sequencing is supposed to reveal the entire genetic makeup of an organism. For infectious disease specialists, the ... Sequencing efforts miss extrachromosomal DNA (phages and plasmids) crucial to bacterias disease causing power. Genomic ... Extensive sequencing of chromosomal DNA has been performed for a variety of pathogenic organisms, but these sequences fail to ... and strands of self-replicating DNA, known as plasmids, often picked up from other bacteria. These phages and plasmids can ...
DNA Sequencing and Analysis of the Low-Ca2+-Response Plasmid pCD1 of Yersinia pestis KIM5. Robert D. Perry, Susan C. Straley, ... DNA Sequencing and Analysis of the Low-Ca2+-Response Plasmid pCD1 of Yersinia pestis KIM5 ... DNA Sequencing and Analysis of the Low-Ca2+-Response Plasmid pCD1 of Yersinia pestis KIM5 ... DNA Sequencing and Analysis of the Low-Ca2+-Response Plasmid pCD1 of Yersinia pestis KIM5 ...
A 22kb region of the 90kb virulence-associated plasmid, pIP1350, of Typhimurium strain C52 was cloned into the mobilizable ... Cloning and expression of plasmid DNA sequences involved in Salmonella serotype typhimurium virulence. ... Transposon Tn5 insertion mutagenesis demonstrated the existence of two DNA sequences in pIP1352 designated VirA and VirB, both ... yielding plasmid pIP1352. This recombinant plasmid restored full virulence to plasmidless strain C53 in a mouse model. ...
... and cloned directly into a plasmid vector. It is only necessary to screen 10-1000 colonies and recombinant DNA is ready for ... Genomic DNA is digested with multiple enzymes cutting outside the fragment to be cloned, selected by electroelution from an ... The use of this technique is demonstrated for the cloning of a sequence from within the human alpha-globin complex that was ... We describe a simple method to directly clone any DNA fragment for which a flanking restriction enzyme map is known. ...
... to correctly predict and emphasize the performance of DNA sequencing techniques as well as in de novo DNA sequencing and its ... On the other hand, the Sanger DNA sequencing method is still considered to be the most reliable; it is a reliable choice for ... face some difficulties in terms of the correct prediction of DNA sequencing outcomes without the implementation of ... virtual modeling to build all possible consensus sequences from smaller DNA fragments. In silico and in vitro experiments were ...
DNA Sequencing.. Nucleotide sequence of a ≈23-kb DNA segment of P. aeruginosa PAK genome was determined by cosmid walking. The ... Plasmid pPT244 (21) containing the PAK fliC gene was used to complement the PAKfliC mutant. The plasmid used for the ... The sequences of the flgK genes in PAK and PAO1 share 86% sequence identity, with two segments corresponding to the first 250 ... Nucleotide Sequence Accession Number.. The nucleotide sequence of the glycosylation island is part of that in GenBank accession ...
Once the plasmid prep is complete, we can perform the sequencing reaction for you. Please indicate clearly which primer(s) you ... This service utilizes magnetic bead technology for high-throughput purification of plasmid DNA from E. coli. It works well for ... For Low Copy Plasmid Prep, please grow your culture in culture tubes using Terrific broth or 2YT broth + antibiotic (NO LB ... For low copy plasmids, please see the special instruction below for growing the cultures. ...
Generates DNA/plasmid plots, virtual agarose gel plots, XY-homology plots - and more! ... analyses DNA for restriction enzymes, homing enzymes and primer sites, dam and dcm methylation sites, finds and translates open ... pDRAW32 DNA analysis software by AcaClone software. pDRAW32 on Windows 7 RC pDRAW32 on OS-X with Wineskin. pDRAW32 on Ubuntu ...
Services , DNA Sequencing , Plasmids. Bacterial plasmids are an essential tool in modern molecular biology research and there ... By undertaking DNA sequencing on your plasmids, we can rapidly provide the key information that enables you to move forward ... DNA Quality. High quality plasmid DNA is essential to achieve the best results. We would recommend a good quality mini-prep, ... We cannot stress too highly the need for plasmid DNA of the highest quality. Even when using commercial plasmid isolation kits ...
Plasmid DNA and DNA sequencing:. Plasmid DNA was isolated using a QIAGEN (Chatsworth, CA) kit. Plasmid DNA was sequenced using ... repaired DNA. Additional libraries were constructed by digesting either MIC DNA or whole-cell unisomic 1 DNA to completion with ... Each 25-μl PCR reaction contained 5 μl of plasmid DNA (5 ng/μl), 2.5 μl of 10× PCR buffer (see recipe above), 2.5 μl of 10 mm ... Nucleotide sequences were determined using an ABI 310 Genetic Analyzer. Sequences were aligned using the BLAST 2 sequences ...
title = "DNA sequence of a plasmid-encoded dihydrofolate reductase",. abstract = "The sequence of the methotrexate-resistant ... DNA sequence of a plasmid-encoded dihydrofolate reductase. In: MGG Molecular & General Genetics. 1981 ; Vol. 181, No. 4. pp. ... Swift G, McCarthy BJ, Heffron F. DNA sequence of a plasmid-encoded dihydrofolate reductase. MGG Molecular & General Genetics. ... DNA sequence of a plasmid-encoded dihydrofolate reductase. / Swift, Galvin; McCarthy, Brian J.; Heffron, Fred. ...
The hybridization of plasmid DNA digested by BsaI, having restriction site within the fosB sequence, demonstrated that the ... The hybridization of plasmid DNA digested by BsaI, having restriction site within the fosB sequence, demonstrated that the ... real-time sequencing. We found that these two plasmids are essentially identical (99.99% nucleotide sequence identity), which ... real-time sequencing. We found that these two plasmids are essentially identical (99.99% nucleotide sequence identity), which ...
Plasmid DNA was sequenced using Pacbio RS II and annotated using RAST.Results: K. pneumoniae RJ119, carrying both blaOXA-48 and ... Plasmid DNA was sequenced using Pacbio RS II and annotated using RAST. Results: K. pneumoniae RJ119, carrying both blaOXA-48 ... blaNDM-1 was located on a 335,317-bp conjugative plasmid, which was a fusion of a blaNDM-1-harboring InA/C plasmid pNDM-US ( ... blaNDM-1 was located on a 335,317-bp conjugative plasmid, which was a fusion of a blaNDM-1-harboring InA/C plasmid pNDM-US ( ...
To mine the gene information relevant to GlcNAc metabolism, the DNA sequences of dasR-dasA-dasBCD-nagB and nagKA in S. ... The dre sequence was proposed by Rigali et al. . Based on MAST analysis, the DNA sequences of the dre motif in the promoter ... The corresponding plasmids were sequenced by Genewiz (Suzhou, China) and the sequences were deposited into the GenBank. The ... d WebLogo of the dre element, and nucleotide sequence of dre motifs in S. verticillus. The DNA binding sequences of DasR were ...
BacteriaPrimersGeneGenomeChromosomesRestrictionPrimerHybridizationGenomic DNANucleotideHigh-throughputDifferent plasmidsEncodeDouble-strandedCopy number plasmidsSpeciesRecombinant plasmidsCDNA2017SimilarityYieldCycle SequencingNext-Generation SequencingAbstractMidiprep KitConjugative plasmidMolecularSensitivity and SpecifiMiniprep KitPurification of DNABindsDetectionConstructsReactionPBluescriptColiTemplateRepeat sequencesCircularSpecific DNA uptake sMethodsReagentStrandsCopiesFacilityReactionsBL21Mammalian cellIncompatibilityMiniprepsTris-EDTA
- Dot-blot DNA hybridizations between pXO2 ORF fragments and total genomic DNA from AWO6 were consistent with the PCR assay results for this isolate and also revealed nine additional ORFs shared between these two bacteria. (biomedcentral.com)
- The presence of pXO2 ORF sequences in other Bacillus species suggests the possibility that certain pXO2 plasmid gene functions may also be present in other closely related bacteria. (biomedcentral.com)
- As a result, the DNA profile of a pathogenic bacteria may be incomplete,' says Vincent Fischetti, head of the Laboratory of Bacterial Pathogenesis and Immunology. (typepad.com)
- Extrachromosomal DNA can include bacteria-infecting viruses, known as phages, and strands of self-replicating DNA, known as plasmids, often picked up from other bacteria. (typepad.com)
- From this information, PCRs identifying novel features of pCT were designed and applied to isolates from several countries, showing that the plasmid has disseminated worldwide in bacteria from humans and animals. (cdc.gov)
- Complete DNA sequences can be used as a platform to develop rapid epidemiologic tools to identify and trace the spread of plasmids in clinically relevant pathogens, thus facilitating a better understanding of their distribution and ability to transfer between bacteria of humans and animals. (cdc.gov)
- CRISPR was discovered in bacteria and function as a defense mechanism against invading viral DNA [ 1 ]. (springer.com)
- In the best-studied naturally competent gram-negative bacteria, Haemophilus influenzae , Haemophilus parainfluenzae , Neisseria gonorrhoeae , and Neisseria meningitidis , double-stranded donor DNA fragments are preferentially bound into a DNase-resistant state if they contain a short recognition sequence abundant in their own genomes, the uptake signal sequence (USS) ( 10 , 11 ). (asm.org)
- StrataPrep midiprep kit purification of plasmid DNA 1 is based on alkaline lysis of bacteria and binding of the liberated plasmid DNA to a glass fiber matrix contained in a midispin cup. (bio-medicine.org)
- Transformation in bacteria is a complex process involving uptake of naked extracellular DNA followed by homologous recombination (HR). Different reproductive barriers have evolved in diverse transformation-competent bacteria, which distinguish in favour of acquisition and recombination of homologous DNA sequences and discriminate against heterologous and potentially hazardous DNA . (prolekare.cz)
- For bacterial species identification, the 16S ribosomal RNA gene can be amplified from all bacteria non-specifically, without amplifying eukaryotic host DNA, or viruses. (nanoporetech.com)
- 132, 912922, 1977), plasmids containing many copies of a protein gene can be used to cause bacteria to make that protein in large quantities. (google.com)
- Specifically, bacteria use restriction enzymes to cut DNA at specific sites. (livestrong.com)
- Researchers can make Escherichia coli bacteria, for example, produce the human insulin protein by inserting the DNA for the protein into bacterial genomes. (livestrong.com)
- Perhaps by targeting the DNA minor groove, we might make antibiotics more effective against VRSA and other drug-resistant bacteria," he says. (innovations-report.com)
- Design and test primers for this sequence using Primer-BLAST. (nih.gov)
- You either have 2 plasmids or 2 primers. (protocol-online.org)
- A standard single reaction consists of one plasmid, PCR fragment, or BAC DNA template sequenced with one primer to produce one chromatogram (A single template sequenced with both forward and reverse primers would comprise two reactions, for example). (albany.edu)
- For common plasmid vectors we provide a number of standard sequencing primers free-of-charge (see list below) or customers can provide custom-designed primers. (albany.edu)
- We provide standard sequencing primers (see below) at no charge. (albany.edu)
- Templates prepared or purified by the facility staff are guaranteed to produce good sequence with standard, reliable primers, barring sequence specific problems (GC rich templates, secondary structure, internal repeats). (upenn.edu)
- I'm a little confused about the notation of the sequencing primers listed on the plasmid map for Invitrogen's pCR 2.1 and 4.0 used in TOPO TA cloning. (protocol-online.org)
- If those primers are actually in forward or reverse orientation on a plasmid map have no importance at all. (protocol-online.org)
- Just a quick note: there are several versions of both the M13 forward and M13 reverse primers, which differ significantly in sequence. (protocol-online.org)
- phage434: M13 primers are included with the kit, and the plasmid map specifies the sequence. (protocol-online.org)
- In total, the sequence of a ca. 7-kb segment of the 9.8-kb plasmid pVM111 was determined by primer walking starting with M13 universal and reverse primers (Stratagene). (asm.org)
- If PCR is performed using Oxford Nanopore's modified primers, sequencing adapters can be attached rapidly following amplification, by chemical ligation. (nanoporetech.com)
- Conservation of plasmid sequences can provide clues about the origin of the pXO2 plasmid and about potentially conserved gene functions. (biomedcentral.com)
- The yopM gene is longer than previously thought (by a sequence encoding two leucine-rich repeats), the ORF upstream of ypkA-yopJ is discussed as a potential Syc gene, and a previously undescribed ORF downstream of yopE was identified as being potentially significant. (pubmedcentralcanada.ca)
- The sequence of the methotrexate-resistant dihydrofolate reductase (DHFR) gene borne by the plasmid R-388 was determined. (elsevier.com)
- This first complete nucleotide sequence of a plasmid carrying two copies of fosB in VRE suggests that the fosB gene can transfer to multiple loci of plasmids by the IS L3 family transposase TnpA, possibly in the form of circular intermediates, leading to the dissemination of high fosfomycin resistance in VRE. (frontiersin.org)
- To mine the gene information relevant to GlcNAc metabolism, the DNA sequences of dasR - dasA - dasBCD - nagB and nagKA in S. verticillus were determined by chromosome walking. (springer.com)
- The sequence was a 93,629-bp plasmid encoding a single antimicrobial drug resistance gene, bla CTX-M-14 . (cdc.gov)
- Bacterial plasmids are key vectors of horizontal gene transfer, mediating the mobilization of genetic material from 1 bacterium to another. (cdc.gov)
- Despite promising reports of the utility of CRISPR-Cas9 for in vivo gene editing, a principal problem in implementing this new process is delivery of high molecular weight DNA into cells. (springer.com)
- 10 ] studied sustained marker gene expression using plasmid DNA in PLGA nanoparticles and liposomes, which concluded that while nanoparticles resulted in a much lower level of gene transfection in vitro, it produced almost two orders of magnitude more successful transfection in vivo than with liposomes. (springer.com)
- Besides a conserved backbone related to pAgK84 of Agrobacterium radiobacter K84, pAMI2 carries a three-gene cluster coding for the protein DmfR, which has sequence similarities to members of the LuxR family of transcription regulators, and two subunits (DmfA1 and DmfA2) of N , N -dimethylformamidase, an enzyme of high substrate specificity that catalyzes the first step in the degradation of DMF. (pubmedcentralcanada.ca)
- The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. (nih.gov)
- Gene sequencing/mapping. (bccresearch.com)
- The organisationally variable forms of the naturally occurring border sequences amongst Ti-plasmid types are differentially responsive to gene products of vir loci concerned with T-strand production. (who.int)
- The interaction of border sequences from different strains of Agrobacterium with vir proteins encoded by various helper plasmids revealed that functional differences do exist amongst vir gene products contained in the type of helper plasmids used. (who.int)
- We propose a DNA-based therapy for pancreatic cancer using a nonviral vector, expressing the diphtheria toxin A chain under the control of the H19 gene regulatory sequences. (hindawi.com)
- Our group previously reported the use of DNA-based therapy for cancer treatment which drives the expression of diphtheria toxin A chain by the regulatory sequence of the H19 gene only into cancer cells [ 5 , 6 ]. (hindawi.com)
- The present work tested the feasibility of using the regulatory sequences of the H19 gene to express the diphtheria toxin A subunit for a DNA-based treatment for pancreatic cancer. (hindawi.com)
- To clarify the gene organization of R64 transfer region, we determined its entire nucleotide sequence. (nii.ac.jp)
- DNA inversions are mediated by rci gene lacated adjacent to shufflon region. (nii.ac.jp)
- nuc gene encoding an EDTA-resistant nuclease and sog gene encoding DNA primase make an operon together with traH-P. This operon is under control of traBC gene. (nii.ac.jp)
- Consequently, the region between the ColE1 origin and the stop codon of the adjacent npt1 gene, which is convergent and confers resistance to kanamycin, was replaced with the corresponding sequence from pBluescript © ( Hellens and Mullineaux 2000 ), a commercially available and widely used cloning vector that has no reported issues with instability ( Alting-Mees and Short 1989 ). (g3journal.org)
- Both the transposon and plasmid were integrated in the proximity of the mecA gene, the latter being flanked by a pair of insertion sequence IS 431 elements. (asm.org)
- The mecA gene encoding PBP2′ has been cloned from a Japanese MRSA strain by exploiting a tobramycin resistance gene which is closely linked to mecA as a selective marker, and its sequence was determined ( 38 ). (asm.org)
- Methods and Results- Targeted deletion of the exon 4 coding sequence in the VDR gene resulted in an increase in myocyte size and left ventricular weight/body weight versus controls both at baseline and following a 7-day infusion of isoproterenol. (ahajournals.org)
- The first tetracycline resistance ( tet ) gene of hybridization class H was detected in 1993 on plasmid pVM111 ( 4 ) from a Pasteurella multocida isolate obtained in 1975 from the tissues of a turkey in California that had died of avian cholera ( 5 ). (asm.org)
- While restriction maps and sequence data for the regions flanking the tetR - tet (H) gene region were available for these plasmids, the corresponding data are still missing for pVM111. (asm.org)
- The telomeres of Tetrahymena ribosomal DNA are not sufficient for stabilizing linear DNA in Xenopus oocytes," Gene 56:313-319 (1987). (freepatentsonline.com)
- In particular, interspecies recombination with heterologous DNA in single cellular organisms could cause gene disruptions and/or disturb sensitive cellular processes, which could in turn have adverse phenotypic consequences. (prolekare.cz)
- As with the whole-genome species ID approach shown in Fig. 1, we have found bead-beating to lyse cells rapidly, yielding DNA with a sufficiently high fragment length for amplification of the 1.5 kb 16S gene. (nanoporetech.com)
- J. R. Sadler and colleagues (Sadler, et al, Gene, 3, 211-232, 1978) were able to prepare from the plasmid pMB9 recombinant plasmids containing from one to four tandem repeats of a synthetic lac operator. (google.com)
- An operator in this context is a DNA segment which controls the expression of a gene. (google.com)
- Later attempts by Sadler and his associates to prepare plasmids containing larger numbers of copies of the operator employed the plasmids containing four operators as starting material (Sadler, et al, Gene, 8, 279-300, 1980). (google.com)
- Before the plasmid gene for drug resistance can be passed, it must be processed for the transfer. (innovations-report.com)
- The final chapter deals with the use of oligonucleotides for the identification and isolation of specific gene sequences. (elsevier.com)
- Scalable throughput and flexibility for virtually any genome, sequencing method, and scale of project. (illumina.com)
- A fast, integrated workflow for a wide range of applications, from human whole-genome sequencing to amplicons, plasmids, and microbial species. (illumina.com)
- Note that sequencing Nextera XT libraries on certain instruments - the HiSeq 1000/2000, HiSeq 1500/2500 in High Output mode, Genome Analyzer, or HiScanSQ systems - also requires one of the following sequencing primer boxes. (illumina.com)
- The whole-genome shotgun sequencing and auto-assembly phase of the Tetrahymena Genome Initiative has recently been completed at The Institute for Genome Research (TIGR). (genetics.org)
- The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. (nih.gov)
- In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. (nih.gov)
- Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. (nih.gov)
- The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. (nih.gov)
- The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead. (nih.gov)
- Genome sequencing/mapping. (bccresearch.com)
- The rapid speed of sequencing attained with modern DNA sequencing technology has been instrumental in the sequencing of complete DNA sequences, or genomes of numerous types and species of life, including the human genome and other complete DNA sequences of many animal, plant, and microbial species. (wikipedia.org)
- Publications] Shoji Fujitani: 'A unique repetitive DNA sequence in the Myxococcus xanthus genome. (nii.ac.jp)
- An earlier version of pGreenII was reported to be unstable, whereby it acquired DNA from the genome of E. coli prior to transfer of plasmid into A. tumefaciens ( Hellens and Mullineaux 2000 ). (g3journal.org)
- The evolutionary pathways of the chlamydial genome and plasmid imply that inheritance of the plasmid is tightly linked with its cognate chromosome. (biomedcentral.com)
- The region, designated mec DNA, is speculated to have originated from the genome of another bacterial species and become integrated into the chromosome of the S. aureus cell in the past. (asm.org)
- Plasmid pLPV-K38 contains a full-length genome of LPV-K38 and was used as a positive control template for PCR analysis. (asm.org)
- Each of the DUS dialects is remarkably conserved within each species and is distributed consistent with a robust Neisseriaceae phylogeny based on core genome sequences. (prolekare.cz)
- We have sequenced the genome of the endangered European eel using the MinION by Oxford Nanopore, and assembled these data using a novel algorithm specifically designed for large eukaryotic genomes. (nanoporetech.com)
- Specific restriction enzymes, called restriction endonucleases, insert segments of DNA into existing segments of DNA, essentially making them a part of an organism's genome. (livestrong.com)
- A plasmid is a section of bacterial DNA that isn't included in the larger genome, or DNA sequence. (livestrong.com)
- They focused on a S. aureus plasmid - a circular loop of double-stranded DNA within the Staph cell separate from the genome - called plW1043 that codes for drug resistance and can be transferred via conjugation ("mating" that involves genetic material passing through a tube from a donor bacterium to a recipient). (innovations-report.com)
- By providing unique sequence from the natural ends of macronuclear chromosomes, Cbs junctions characterized in the work reported here will serve as useful sequence tags for relating macro- and micronuclear genetic, physical, and sequence maps. (genetics.org)
- Analysis of chlamydial plasmids and their cognate chromosomes was undertaken to provide insights into the evolutionary relationship between chromosome and plasmid. (biomedcentral.com)
- Phylogenetic analysis of the plasmids and chromosomes are fully congruent. (biomedcentral.com)
- He has found two circular chromosomes or plasmids in Rhodobacter with Kaplan S in 1989. (wikipedia.org)
- DNA sequence of a plasmid pEIB1 associated with virulence from the marine fish pathogen Vibrio anguillarum was determined using the methods of restriction endonuclease digestion, subcloning, and primer walking. (eurekamag.com)
- We describe a simple method to directly clone any DNA fragment for which a flanking restriction enzyme map is known. (ox.ac.uk)
- The hybridization of plasmid DNA digested by Bsa I, having restriction site within the fosB sequence, demonstrated that the presence of multiple copies of fosB per plasmid is common. (frontiersin.org)
- A restriction enzyme or restriction endonuclease is an enzyme that cleaves DNA into fragments at or near specific recognition sites within the molecule known as restriction sites . (wikipedia.org)
- Restriction enzymes are commonly classified into five types, which differ in their structure and whether they cut their DNA substrate at their recognition site, or if the recognition and cleavage sites are separate from one another. (wikipedia.org)
- To cut DNA, all restriction enzymes make two incisions, once through each sugar-phosphate backbone (i.e. each strand) of the DNA double helix. (wikipedia.org)
- In the 1960s, it was shown in work done in the laboratories of Werner Arber and Matthew Meselson that the restriction is caused by an enzymatic cleavage of the phage DNA, and the enzyme involved was therefore termed a restriction enzyme. (wikipedia.org)
- The restriction enzymes studied by Arber and Meselson were type I restriction enzymes, which cleave DNA randomly away from the recognition site. (wikipedia.org)
- Restriction enzymes of this type are more useful for laboratory work as they cleave DNA at the site of their recognition sequence and are the most commonly used as a molecular biology tool. (wikipedia.org)
- Later, Daniel Nathans and Kathleen Danna showed that cleavage of simian virus 40 (SV40) DNA by restriction enzymes yields specific fragments that can be separated using polyacrylamide gel electrophoresis , thus showing that restriction enzymes can also be used for mapping DNA. (wikipedia.org)
- Restriction enzymes recognize a specific sequence of nucleotides and produce a double-stranded cut in the DNA. (wikipedia.org)
- The quality of the DNA ensures robust performance in applications such as sequencing, restriction analyses, and cloning. (bio-medicine.org)
- The isolated DNA can be used directly in standard applications, including automated sequencing, PCR, restriction analysis, and cloning. (bio-medicine.org)
- The resulting DNA can be used for applications such as restriction digestion, cloning, PCR, site-directed mutagenesis, manual and automated fluorescent sequencing, and mammalian transfection. (bio-medicine.org)
- Plasmid DNA is ready for immediate use in restriction enzyme digestion (Figure 1) cloning, PCR, transcription, conventional and automated sequencing (Figure 2) . (sigmaaldrich.com)
- Plasmid DNA purified with GenElute Plasmid Miniprep Kit is suitable for restriction enzyme digestions. (sigmaaldrich.com)
- Restriction digestions of pUC19 purified from JM109 using the GenElute Plasmid Miniprep Kit. (sigmaaldrich.com)
- Pure plasmid DNA suitable for restriction mapping, PCR analysis, cloning experiments, and sequence analysis was obtained by alkaline lysis with subsequent purification by affinity chromatography on Midi columns (Qiagen, Hilden, Germany) ( 9 ). (asm.org)
- Genetic reagents for generating plasmids containing multiples of a DNA segment are prepared by modification of the restriction sites of plasmid rings. (google.com)
- The plasmid rings with modified restriction sites are also used in linear form to obtain head-to-tail joining of multiple DNA segments into plasmid rings for further cloning and/or expression of the DNA segment-directed protein. (google.com)
- The critical restriction site sequences of the modified plasmid rings can be prepared as a reagent which permits the sequence to be introduced into any plasmid. (google.com)
- 5. A genetic reagent for generating plasmids containing multiple copies of DNA segments, comprising a pBR322 plasmid characterized by containing a sequence of three contiguous restriction sites in which the first and third sites are Ava I sites having non-symmetrically arranged base pairs, the second site is an EcoR I site, and said plasmid ring contains no other sites cleavable by either Ava I EcoR I restriction endonucleases. (google.com)
- Restriction enzymes cut DNA at specific sites. (livestrong.com)
- Bacterial species use restriction enzymes to help protect themselves against foreign DNA. (livestrong.com)
- Restriction enzymes literally "restrict" the ability of the virus to infect a bacterial cell by recognizing and chopping up viral DNA before it can do damage to the cell. (livestrong.com)
- Scientists can also use restriction enzymes to map plasmids, through a process called restriction mapping. (livestrong.com)
- Scientists can use restriction enzymes to break a plasmid into chunks, and then test the chunks to determine the DNA sequence of each. (livestrong.com)
- Restriction enzymes of this type are more useful for laboratory work as they cleave DNA at the site of their recognition sequence. (wikipedia.org)
- EcoRI digestion produces "sticky" ends, whereas SmaI restriction enzyme cleavage produces "blunt" ends: Recognition sequences in DNA differ for each restriction enzyme, producing differences in the length, sequence and strand orientation (5' end or 3' end) of a sticky-end "overhang" of an enzyme restriction. (wikipedia.org)
- Nothing - bad primer, loss of DNA, bad cycling conditions. (protocol-online.org)
- PCR was performed using template DNA from 11 Bacillus isolates that vary in relatedness to B. anthracis with primer sets designed to amplify DNA fragments from 47 different pXO2 ORFs. (biomedcentral.com)
- increased concentration, but reduced total volume per reaction (i.e. one template sequenced with one primer). (dnaseq.co.uk)
- Cycle Sequencing was performed using 500 ng template, a T7 sequencing primer and ABI BigDye™ terminator chemistry. (sigmaaldrich.com)
- This option is best when your template/primer combinations are below the threshold for discounted economy sequencing. (albany.edu)
- For primer walking projects we recommend that a substantial amount of DNA (nominally 4-5 μg per kb for each direction) be treated this way and be used for the whole project. (upenn.edu)
- Plasmid, M13 primer that is 5' to the insert site is called the reverse primer and the M13 primer that is 3' to the insert is called the forward primer. (protocol-online.org)
- PCR, forward primer has the same sequence as the 5' to 3' template and is 5' to the region being amplifying. (protocol-online.org)
- While the reverse primer sequence is complementary to the 5' to 3' sequence and is 3' to the region being amplified. (protocol-online.org)
- Seems opposite of sequencing primer notation. (protocol-online.org)
- I used the forward sequencing primer the first time and the sequence matched my reference but when I sequenced with the T3 priming site primer (5' to the insert) I got the complementary sequence to my reference. (protocol-online.org)
- Primer sequences with M and Y - how to order? (protocol-online.org)
- Fragments of cDNA clones were sequenced using a T3 primer from the 5′ end with MegaBase1000 DNA sequencer. (hindawi.com)
- Primer pairs LPR1/LPR2 and LPR3/LPR4 designed to detect LPV regulatory region sequences are given in Table 1 . (asm.org)
- PCR analysis of PBMC DNA with primer pair LPR3/LPR4 yielded a robust PCR-amplified band that was estimated to be about 550 bp in size from the K661 lane A sample, but not from the other samples (Fig. 2 ). (asm.org)
- The same approach with the primer combination sul1 and str2 (the sequence of the forward primer was from the 5′ end of sul2 , and the sequence of the reverse primer was from the 3′ end of strA ) ( 10 ) and pVM111 DNA as the template resulted in an amplicon of ca. 3.5 kb. (asm.org)
- David Hi David, we use (successfully) the following protocoll in our lab: · Master-Mix for every sequence: 4 × 50 ng/kb plasmid 4 × 2 pmol primer 4 × ad 6 µl H2O · Make aliquots of 6 µl each · add 2 µl Reactionsmix (e. g. (bio.net)
- If the strands reassociate to yield an unpaired repeat on the primer strand, the net result is an addition of repeats (following a second round of DNA replication). (asm.org)
- The plasmids harboring bla OXA-48 and bla NDM-1 were analyzed through conjugation experiments, S1-nuclease pulsed-field gel electrophoresis, and hybridization with specific probes. (frontiersin.org)
- Strain PT23 of Pseudomonas syringae pv, tomato contains four native plasmids, designated A, B, C, and D. By DNA hybridization of genomic and plasmid DNA digests from the wild type and a plasmid-cured strain, we determined that c. 61 kb (c. 74%) of pPT23B is repeated in pPT23A and only c. 17 kb (c. 21%) is in single copy in strain PT23. (unavarra.es)
- By DNA hybridization with the origin of replication from a native plasmid of P. syringae pv. (unavarra.es)
- Direct cloning of specific genomic DNA sequences in plasmid libraries following fragment enrichment. (ox.ac.uk)
- Genomic DNA is digested with multiple enzymes cutting outside the fragment to be cloned, selected by electroelution from an agarose gel, and cloned directly into a plasmid vector. (ox.ac.uk)
- Genomic DNA Prep: 96well block format only. (ucberkeleydnasequencing.com)
- We compared commercial kits for extraction of genomic DNA from the Gram-negative bacterium Klebsiella pneumoniae for subsequent Miseq sequencing. (rki.de)
- Indeed, the genomic DNA of A. actinomycetemcomitans was identified in atherosclerotic lesions ( 14 ), suggesting a possible role of this bacterium in cardiovascular disease. (asm.org)
- We regularly use this kit to isolate genomic DNA of bacterial transposon libraries. (biocompare.com)
- The tool works with standard single letter nucleotide or protein codes including ambiguities and can match Prosite patterns in protein sequences. (nih.gov)
- We found that these two plasmids are essentially identical (99.99% nucleotide sequence identity), which proved the possibility of interspecies transmission. (frontiersin.org)
- bla OXA-48 was located on a 61,748-bp IncL/M conjugative plasmid, which displayed overall nucleotide identity (99%) to pKPN-E1-Nr.7. (frontiersin.org)
- DnaSP, DNA Sequence Polymorphism, is a software package for the analysis of nucleotide polymorphism from aligned DNA sequence data. (pcwin.com)
- Publications] Nobuhisa Furuya: 'Nucleotide sequence and function of the oriT operon in IncI1 plasmid R64. (nii.ac.jp)
- Publications] Su-Ryang Kim: 'Nucleotide sequence of the R721 shufflon. (nii.ac.jp)
- There were a number of sequence ambiguities generated from each template, starting arbitrarily from nucleotide 20: one for a StrataPrep midiprep standard, one for a StrataPrep midiprep endotoxin-free, zero for CsCl density gradient centrifugation, and four for anion exchange. (bio-medicine.org)
- Non-replicating vectors containing a nucleotide sequence coding for an F protein of respiratory syncytial virus (RSV) and a promoter for such sequence, preferably a cytomegalovirus promoter, are described for in vivo immunization. (google.es)
- Such non-replicating vectors, including plasmids, also may contain a further nucleotide sequence located adjacent to the RSV F protein encoding sequence to enhance the immunoprotective ability of the RSV F protein when expressed in vivo. (google.es)
- 2. The composition of claim 1 wherein said first nucleotide sequence encodes a full-length RSV F protein having SEO ID No: 2. (google.es)
- 3. The composition of claim 1 wherein said first nucleotide sequence encodes a RSV F protein from which the transmembrane region is absent and having SEQ ID No: 4. (google.es)
- 5. The composition of claim 1 wherein said second nucleotide sequence is that of rabbit β-globin intron II. (google.es)
- The impact of individual single nucleotide transversions in DUS on meningococcal transformation and on DNA binding and uptake is analysed. (prolekare.cz)
- Eggs injected with one of two different plasmids yielded no transgenic fish, but 7-25% (19 of 115 overall) of the eggs injected with the other plasmid transmitted the injected sequences to their offspring (F1). (pnas.org)
- Bacterial plasmids are an essential tool in modern molecular biology research and there are many different plasmids used for various purposes, including basic cloning, expression in a range of cell types and mutagenesis of inserts. (dnaseq.co.uk)
- To describe the genetic environment, transferability, and antibiotic susceptibility of one clinical Klebsiella pneumoniae isolate harboring both bla OXA-48 and bla NDM-1 on different plasmids from a Chinese hospital. (frontiersin.org)
- ColV plasmids, which encode ColV production, typically range in size from 80 to 180 kb ( 53 ) and encode traits such as aerobactin production ( 51 ) and complement resistance ( 31 ). (asm.org)
- The CRISPR-Cas9 system is typically delivered to cells as a single large plasmid or multiple smaller plasmids that encode a target sequence, a CRISPR guide, and Cas9. (springer.com)
- A chapter specifically tackles the protocols of DNA synthesis using double-stranded plasmid DNA as a template. (elsevier.com)
- The inverted repeat palindrome is also a sequence that reads the same forward and backward, but the forward and backward sequences are found in complementary DNA strands (i.e., of double-stranded DNA), as in GTATAC (GTATAC being complementary to CATATG). (wikipedia.org)
- The degree of similarity between the DUS dialect of the recipient species and the donor DNA directly correlates with the level of transformation and DNA binding and uptake. (prolekare.cz)
- The antigenic potential of decorin binding protein A (DbpA) was evaluated in serodiagnosis of human Lyme borreliosis (LB). The dbpA was cloned and sequenced from the three pathogenic Borrelia species common in Europe. (asm.org)
- Chlamydiaceae species have varying inclusion morphology, varying extrachromosomal plasmid content, and varying sulfadiazine resistance. (wikipedia.org)
- An expressed sequence tag or EST is a short sub-sequence of a cDNA (complementary DNA) sequence. (pcwin.com)
- Unidirectional cDNA libraries were constructed using the pBluescript II plasmid system. (hindawi.com)
- In the NOD model, rIL-10 protein, IL-10 fusion protein, expression plasmid, or adoptive transfer with IL-10-cDNA-transduced islet-specific T cell clones have all been shown to prevent the onset of spontaneous diabetes ( 6 , 7 , 8 , 9 ). (jimmunol.org)
- Find regions of similarity between this sequence and other sequences using BLAST. (nih.gov)
- This method was chosen to detect sequences with potential similarity to pXO2 because it is rapid and the reaction products can be readily sequenced. (biomedcentral.com)
- Publications] Nobuhisa Furuya: 'Determination of the nick site at oriT of IncI1 plasmid R64;Global similarity of oriT structures of IncI1 and IncP plasmids. (nii.ac.jp)
- Alternatively, we are happy to process your preps for you to yield DNA of appropriate quality (conditions apply in terms of copy number and plasmid size). (dnaseq.co.uk)
- Nearly 350 g of high-copy number pBluescript II SK(+) plasmid DNA was also recovered from a 100-ml overnight bacterial culture using a single midispin cup, while the low-copy number pCMV gal plasmid yield from a 200-ml overnight culture was 250 g ( Figure 3 ). (bio-medicine.org)
- bla NDM-1 was located on a 335,317-bp conjugative plasmid, which was a fusion of a bla NDM-1 -harboring InA/C plasmid pNDM-US (140,825 bp, 99% identity) and an IncFIB plasmid pKPN-c22 (178,563 bp, 99% identity). (frontiersin.org)
- R64 is a 122-kb conjugative plasmid belonging to the incompatibility group I1. (nii.ac.jp)
- It is only necessary to screen 10-1000 colonies and recombinant DNA is ready for immediate molecular analysis without further subcloning. (ox.ac.uk)
- These enzymes are routinely used for DNA modification in laboratories, and they are a vital tool in molecular cloning . (wikipedia.org)
- It is a molecular biology software that can view and assemble sequences in contigs. (pcwin.com)
- It is a molecular biology software that can view and assemble sequences in contigs.Features: end clip, export sequences, assemblies, code detection. (pcwin.com)
- In October 2006 a new variant of C. trachomatis was described in Sweden that evaded several of the then current commercial molecular diagnostic tests for detecting the microorganism, which were based on the presence of specific plasmid sequences [ 8 ]. (biomedcentral.com)
- High-quality plasmid DNA for all demanding molecular biology applic. (bio-medicine.org)
- A major rate-limiting step in this process can be the extraction of sufficiently pure, high-molecular weight DNA from the sample, and so to address this limitation we evaluated several different methods of cell lysis, including both enzymatic and physical approaches. (nanoporetech.com)
- QuickLyse Miniprep Kit ( For ultra-fast sequencing grad. (bio-medicine.org)
- The QIAGEN QuickLyse Miniprep Kit uses one-step lysis technology for ultra-fast preparation of up to 15 g plasmid DNA. (bio-medicine.org)
- The QuickLyse Miniprep Kit yields s ufficient plasmid DNA for any downstream application, including automated sequencing (Figure 2). (bio-medicine.org)
- GenElute™ Plasmid Miniprep Kit offers a simple, rapid, and cost-effective method for isolating up to 20 µg plasmid DNA from E. coli cultures. (sigmaaldrich.com)
- 600 bases of sequence from pUC-TMV using GenElute Plasmid Miniprep Kit. (sigmaaldrich.com)
- Plasmid DNA binds to the column membrane and is eluted after a single wash step. (bio-medicine.org)
- Upon centrifugation, plasmid DNA binds to the glass fiber matrix in the midispin cup. (bio-medicine.org)
- This occurs when a protein called the Nicking Enzyme of Staphylococci, or NES, binds with its active area, known as the relaxase region, to the donor cell plasmid. (innovations-report.com)
- these can fluoresce and interfere with the detection of sequencing products. (albany.edu)
- Easy to use aligner software for DNA sequence contig assembly, contig editing, and mutation detection for Windows. (pcwin.com)
- The plasmid from the new Swedish variant has a 377 bp deletion in the first predicted coding sequence, abolishing the site used for PCR detection, resulting in negative diagnosis. (biomedcentral.com)
- Using either of two plasmid constructs, we injected uncut DNA into fertilized eggs at the one- or two-cell stage. (pnas.org)
- To test this we used plasmid constructs that were identical, except that the UTR DNA sequence was the wild‐type (pRL11), LV‐1 or LV‐2. (nih.gov)
- I didn't know if this would work, but the potential of programmable DNA constructs was unbelievable," he says. (sciencemag.org)
- Now he could design, build, and test DNA constructs to control the cell's machinery through standard biochemical operations like transcription, translation, or post-translation processes. (sciencemag.org)
- How much DNA is in each reaction? (protocol-online.org)
- If you clean up your own sequencing reactions, this is another point that could ruin your reaction. (protocol-online.org)
- I percipitate my 10ul reaction sample by first diluting it with 90ul of sterile distiled water and 1ul Pellet paint (basically coloured dextran that helps pulls down the DNA into a nice colour pellet). (protocol-online.org)
- Once the plasmid prep is complete, we can perform the sequencing reaction for you. (ucberkeleydnasequencing.com)
- We consider plasmids of up to about 20kb in length to be 'standard' and we recommend our standard sequencing reaction protocol for such plasmids. (dnaseq.co.uk)
- Tris-EDTA buffer), which can inhibit the sequencing reaction. (dnaseq.co.uk)
- If you cannot achieve the stated template concentration, we can compensate by adding more template volume (up to 6 µl) per reaction, but note that we have observed a decline in sequencing quality with DNAs less than 1/4 the recommended concentration, regardless of this adjustment. (albany.edu)
- If your plasmid is larger than this, you should select our "Large Template" option, which is specifically designed to give improved results with large plasmids. (dnaseq.co.uk)
- Template DNA can be prepared and/or purified by the facility staff. (upenn.edu)
- This treatment improves quality of template, producing more reliable sequence and longer reads. (upenn.edu)
- To ensure optimum template concentration, we suggest you to determine the concentration of your template DNA prior to shipment. (genscript.com)
- R64 shufflon consists of four DNA segments, which are flanked and separated by seven 19-bp repeat sequences. (nii.ac.jp)
- The mec DNA was found to be 51,669 bp long, including terminal inverted repeats of 27 bp and a characteristic pair of direct repeat sequences of 15 bp each: one is situated in the right extremity of mec DNA, and the other is situated outside the mec DNA and abuts the left boundary of mec DNA. (asm.org)
- The telomere maintenance mechanism in most immortalized cells and tumor cells utilizes the ribonucleoprotein enzyme telomerase, which compensates for sequential telomere shortening at each cell division by catalyzing the addition of repeat sequences ( 15 ). (asm.org)
- DNA sequencing has developed dramatically in recent years, with many methods now available. (selectscience.net)
- The advent of rapid DNA sequencing methods has greatly accelerated biological and medical research and discovery. (wikipedia.org)
- The first DNA sequences were obtained in the early 1970s by academic researchers using laborious methods based on two-dimensional chromatography . (wikipedia.org)
- DNA sequencing may be used along with DNA profiling methods for forensic identification and paternity testing . (wikipedia.org)
- Plasmid DNA purified by a StrataPrep standard midiprep, a StrataPrep endotoxin-free midiprep, twice-banded CsCl density gradient centrifugation, and anion exchange methods were sequenced at Sequetech Corporation. (bio-medicine.org)
- LipoTAXI transfection reagent 3,4 and calcium phosphate 5 reagent methods were used to transfect StrataPrep endotoxin-free purified DNA into Cos-7, DU 145, and JAR cell lines and resulted in higher transfection efficiencies, compared to anion exchange enndotoxin-free purified plasmid DNA ( Figure 5 ). (bio-medicine.org)
- on both strands, and CC protects the DNA from cleavage by the CfrBI endonuclease. (genome.jp)
- NES then cuts, or "nicks," one strand of the double helix so that it separates into two single strands of DNA. (innovations-report.com)
- After the two strands are replicated, NES reforms the plasmid in both cells, creating two drug-resistant Staph cells that are ready to spread their misery further. (innovations-report.com)
- We realized that a compound that could block this groove, prevent the NES loops from attaching and inhibit the cleaving of the plasmid DNA into single strands could potentially stop conjugal transfer of drug resistance altogether," Edwards says. (innovations-report.com)
- Two single strands of a replicating DNA molecule are shown, with each repeat unit indicated by a rectangle. (asm.org)
- Release studies revealed that most of the DNA was released within the first 24 h and corresponded to ~ 2-3 plasmid copies released per nanoparticle. (springer.com)
- In particular, this invention is concerned with the preparation of plasmids containing many tandem copies of a selected DNA segment. (google.com)
- In addition, the multiple copies of the DNA segments have commercial value in themselves, for example as reagents for recombinant DNA processes. (google.com)
- The DNA sequencing facility at CFG offers DNA sequencing services for a wide range of templates. (albany.edu)
- If used for sequencing, the Facility guarantees the results, i.e. we will repurify and repeat sequencing at no additional charge until successful sequence is obtained. (upenn.edu)
- The facility also offers reliable treatment of marginal or bad templates to make them suitable for automated sequencing. (upenn.edu)