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.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.R Factors: A class of plasmids that transfer antibiotic resistance from one bacterium to another by conjugation.Bacteriocin Plasmids: Plasmids encoding bacterial exotoxins (BACTERIOCINS).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.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Genes, Bacterial: The functional hereditary units of BACTERIA.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.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 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.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.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.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.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)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.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.Chromosomes, Bacterial: Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.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).DNA Replication: The process by which a DNA molecule is duplicated.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.Bacterial Proteins: Proteins found in any species of bacterium.Restriction Mapping: Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.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)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.Sequence Analysis, DNA: A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.Plant Tumor-Inducing Plasmids: Plasmids coding for proteins which induce PLANT TUMORS. The most notable example of a plant tumor inducing plasmid is the Ti plasmid found associated with AGROBACTERIUM TUMEFACIENS.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.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)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).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)Electrophoresis, Agar Gel: Electrophoresis in which agar or agarose gel is used as the diffusion medium.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.Tetracycline: A naphthacene antibiotic that inhibits AMINO ACYL TRNA binding during protein synthesis.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.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.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.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.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.Anti-Bacterial Agents: Substances that reduce the growth or reproduction of BACTERIA.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.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.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.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.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.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.Escherichia coli Proteins: Proteins obtained from ESCHERICHIA COLI.Enterobacteriaceae: A family of gram-negative, facultatively anaerobic, rod-shaped bacteria that do not form endospores. Its organisms are distributed worldwide with some being saprophytes and others being plant and animal parasites. Many species are of considerable economic importance due to their pathogenic effects on agriculture and livestock.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.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.Tetracycline Resistance: Nonsusceptibility of bacteria to the action of TETRACYCLINE which inhibits aminoacyl-tRNA binding to the 30S ribosomal subunit during protein synthesis.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)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.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.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.Chromosome Mapping: Any method used for determining the location of and relative distances between genes on a chromosome.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.Salmonella: A genus of gram-negative, facultatively anaerobic, rod-shaped bacteria that utilizes citrate as a sole carbon source. It is pathogenic for humans, causing enteric fevers, gastroenteritis, and bacteremia. Food poisoning is the most common clinical manifestation. Organisms within this genus are separated on the basis of antigenic characteristics, sugar fermentation patterns, and bacteriophage susceptibility.Pseudomonas: A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in nature. Some species are pathogenic for humans, animals, and plants.Streptomycin: An antibiotic produced by the soil actinomycete Streptomyces griseus. It acts by inhibiting the initiation and elongation processes during protein synthesis.Klebsiella pneumoniae: Gram-negative, non-motile, capsulated, gas-producing rods found widely in nature and associated with urinary and respiratory infections in humans.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).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.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).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).Cell Line: Established cell cultures that have the potential to propagate indefinitely.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.Penicillinase: A beta-lactamase preferentially cleaving penicillins. (Dorland, 28th ed) EC 3.5.2.-.Gene Expression Regulation, Bacterial: Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria.DNA, Viral: Deoxyribonucleic acid that makes up the genetic material of viruses.Drug Resistance, Bacterial: The ability of 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).Genome, Bacterial: The genetic complement of a BACTERIA as represented in its DNA.Drug Resistance, Multiple, Bacterial: The ability of bacteria to resist or to become tolerant to several structurally and functionally distinct drugs simultaneously. This resistance may be acquired through gene mutation or foreign DNA in transmissible plasmids (R FACTORS).Chloramphenicol: An antibiotic first isolated from cultures of Streptomyces venequelae in 1947 but now produced synthetically. It has a relatively simple structure and was the first broad-spectrum antibiotic to be discovered. It acts by interfering with bacterial protein synthesis and is mainly bacteriostatic. (From Martindale, The Extra Pharmacopoeia, 29th ed, p106)Lactose Factors: Plasmids which determine the ability of a bacterium to ferment lactose.Electrophoresis, Gel, Pulsed-Field: Gel electrophoresis in which the direction of the electric field is changed periodically. This technique is similar to other electrophoretic methods normally used to separate double-stranded DNA molecules ranging in size up to tens of thousands of base-pairs. However, by alternating the electric field direction one is able to separate DNA molecules up to several million base-pairs in length.Salmonella typhimurium: A serotype of Salmonella enterica that is a frequent agent of Salmonella gastroenteritis in humans. It also causes PARATYPHOID FEVER.Nucleic Acid Conformation: The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.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.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.Microbial Sensitivity Tests: Any tests that demonstrate the relative efficacy of different chemotherapeutic agents against specific microorganisms (i.e., bacteria, fungi, viruses).Chloramphenicol Resistance: Nonsusceptibility of bacteria to the action of CHLORAMPHENICOL, a potent inhibitor of protein synthesis in the 50S ribosomal subunit where amino acids are added to nascent bacterial polypeptides.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.Phenotype: The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.DNA, Fungal: Deoxyribonucleic acid that makes up the genetic material of fungi.Gene Order: The sequential location of genes on a chromosome.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 126.96.36.199.Phylogeny: The relationships of groups of organisms as reflected by their genetic makeup.Enterococcus faecalis: A species of gram-positive, coccoid bacteria commonly isolated from clinical specimens and the human intestinal tract. Most strains are nonhemolytic.Nebramycin: A complex of antibiotic substances produced by Streptomyces tenebrarius.Molecular Weight: The sum of the weight of all the atoms in a molecule.Kanamycin Resistance: Nonsusceptibility of bacteria to the antibiotic KANAMYCIN, which can bind to their 70S ribosomes and cause misreading of messenger RNA.Bacteriophages: Viruses whose hosts are bacterial cells.Escherichia coli Infections: Infections with bacteria of the species ESCHERICHIA COLI.Ampicillin: Semi-synthetic derivative of penicillin that functions as an orally active broad-spectrum antibiotic.Deoxyribonuclease EcoRI: One of the Type II site-specific deoxyribonucleases (EC 188.8.131.52). 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.-.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.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 184.108.40.206.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.Trimethoprim: A pyrimidine inhibitor of dihydrofolate reductase, it is an antibacterial related to PYRIMETHAMINE. It is potentiated by SULFONAMIDES and the TRIMETHOPRIM, SULFAMETHOXAZOLE DRUG COMBINATION is the form most often used. It is sometimes used alone as an antimalarial. TRIMETHOPRIM RESISTANCE has been reported.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.Genes, Viral: The functional hereditary units of VIRUSES.Mercury: A silver metallic element that exists as a liquid at room temperature. It has the atomic symbol Hg (from hydrargyrum, liquid silver), atomic number 80, and atomic weight 200.59. Mercury is used in many industrial applications and its salts have been employed therapeutically as purgatives, antisyphilitics, disinfectants, and astringents. It can be absorbed through the skin and mucous membranes which leads to MERCURY POISONING. Because of its toxicity, the clinical use of mercury and mercurials is diminishing.Trimethoprim Resistance: Nonsusceptibility of bacteria to the action of TRIMETHOPRIM.Integrons: DNA elements that include the component genes and insertion site for a site-specific recombination system that enables them to capture mobile gene cassettes.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.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.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.Spiroplasma citri: The type species of gram-negative bacteria in the genus SPIROPLASMA, family SPIROPLASMATACEAE, causing citrus stubborn disease.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.DNA Helicases: Proteins that catalyze the unwinding of duplex DNA during replication by binding cooperatively to single-stranded regions of DNA or to short regions of duplex DNA that are undergoing transient opening. In addition DNA helicases are DNA-dependent ATPases that harness the free energy of ATP hydrolysis to translocate DNA strands.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.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.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.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.Rhodococcus equi: A species of RHODOCOCCUS found in soil, herbivore dung, and in the intestinal tract of cows, horses, sheep, and pigs. It causes bronchopneumonia in foals and can be responsible for infection in humans compromised by immunosuppressive drug therapy, lymphoma, or AIDS.Viral Proteins: Proteins found in any species of virus.Tellurium: Tellurium. An element that is a member of the chalcogen family. It has the atomic symbol Te, atomic number 52, and atomic weight 127.60. It has been used as a coloring agent and in the manufacture of electrical equipment. Exposure may cause nausea, vomiting, and CNS depression.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.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.Gene Expression: The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.Bacillus subtilis: A species of gram-positive bacteria that is a common soil and water saprophyte.Coliphages: Viruses whose host is Escherichia coli.Penicillin Resistance: Nonsusceptibility of an organism to the action of penicillins.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.Pseudomonas aeruginosa: A species of gram-negative, aerobic, rod-shaped bacteria commonly isolated from clinical specimens (wound, burn, and urinary tract infections). It is also found widely distributed in soil and water. P. aeruginosa is a major agent of nosocomial infection.Mutagenesis: Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.Shigella sonnei: A lactose-fermenting bacterium causing dysentery.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.Gentamicins: A complex of closely related aminoglycosides obtained from MICROMONOSPORA purpurea and related species. They are broad-spectrum antibiotics, but may cause ear and kidney damage. They act to inhibit PROTEIN BIOSYNTHESIS.Interspersed Repetitive Sequences: Copies of transposable elements interspersed throughout the genome, some of which are still active and often referred to as "jumping genes". There are two classes of interspersed repetitive elements. Class I elements (or RETROELEMENTS - such as retrotransposons, retroviruses, LONG INTERSPERSED NUCLEOTIDE ELEMENTS and SHORT INTERSPERSED NUCLEOTIDE ELEMENTS) transpose via reverse transcription of an RNA intermediate. Class II elements (or DNA TRANSPOSABLE ELEMENTS - such as transposons, Tn elements, insertion sequence elements and mobile gene cassettes of bacterial integrons) transpose directly from one site in the DNA to another.Chromosome Deletion: Actual loss of portion of a chromosome.Deoxyribonuclease HindIII: One of the Type II site-specific deoxyribonucleases (EC 220.127.116.11). 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.-.Staphylococcus aureus: Potentially pathogenic bacteria found in nasal membranes, skin, hair follicles, and perineum of warm-blooded animals. They may cause a wide range of infections and intoxications.Bacteria: One of the three domains of life (the others being Eukarya and ARCHAEA), also called Eubacteria. They are unicellular prokaryotic microorganisms which generally possess rigid cell walls, multiply by cell division, and exhibit three principal forms: round or coccal, rodlike or bacillary, and spiral or spirochetal. Bacteria can be classified by their response to OXYGEN: aerobic, anaerobic, or facultatively anaerobic; by the mode by which they obtain their energy: chemotrophy (via chemical reaction) or PHOTOTROPHY (via light reaction); for chemotrophs by their source of chemical energy: CHEMOLITHOTROPHY (from inorganic compounds) or chemoorganotrophy (from organic compounds); and by their source for CARBON; NITROGEN; etc.; HETEROTROPHY (from organic sources) or AUTOTROPHY (from CARBON DIOXIDE). They can also be classified by whether or not they stain (based on the structure of their CELL WALLS) with CRYSTAL VIOLET dye: gram-negative or gram-positive.Bacterial Toxins: Toxic substances formed in or elaborated by bacteria; they are usually proteins with high molecular weight and antigenicity; some are used as antibiotics and some to skin test for the presence of or susceptibility to certain diseases.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.Thiamphenicol: A methylsulfonyl analog of CHLORAMPHENICOL. It is an antibiotic and immunosuppressive agent.Enterobacteriaceae Infections: Infections with bacteria of the family ENTEROBACTERIACEAE.Klebsiella Infections: Infections with bacteria of the genus KLEBSIELLA.Hemolysin Factors: Plasmids controlling the synthesis of hemolysin by bacteria.RNA Phages: Bacteriophages whose genetic material is RNA, which is single-stranded in all except the Pseudomonas phage phi 6 (BACTERIOPHAGE PHI 6). All RNA phages infect their host bacteria via the host's surface pili. Some frequently encountered RNA phages are: BF23, F2, R17, fr, PhiCb5, PhiCb12r, PhiCb8r, PhiCb23r, 7s, PP7, Q beta phage, MS2 phage, and BACTERIOPHAGE PHI 6.Clostridium perfringens: The most common etiologic agent of GAS GANGRENE. It is differentiable into several distinct types based on the distribution of twelve different toxins.Virus Replication: The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle.Ampicillin Resistance: Nonsusceptibility of a microbe to the action of ampicillin, a penicillin derivative that interferes with cell wall synthesis.Virulence Factors: Those components of an organism that determine its capacity to cause disease but are not required for its viability per se. Two classes have been characterized: TOXINS, BIOLOGICAL and surface adhesion molecules that effect the ability of the microorganism to invade and colonize a host. (From Davis et al., Microbiology, 4th ed. p486)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.Klebsiella oxytoca: A species of gram-negative bacteria causing URINARY TRACT INFECTIONS and SEPTICEMIA.beta-Lactam Resistance: Nonsusceptibility of bacteria to the action of the beta-lactam antibiotics. Mechanisms responsible for beta-lactam resistance may be degradation of antibiotics by BETA-LACTAMASES, failure of antibiotics to penetrate, or low-affinity binding of antibiotics to targets.Genes, Fungal: The functional hereditary units of FUNGI.Serotyping: Process of determining and distinguishing species of bacteria or viruses based on antigens they share.
Human HIST1H2BH Gene ORF cDNA clone expression plasmid, N-HA tag | SinoBiological... expression plasmid, mutiple tags. Optimized for high expression in mammalian cells. Save up to 60%. ... Each tube contains lyophilized plasmid.. Storage. :. The lyophilized plasmid can be stored at room temperature for three months ... Human HIST1H2BH Gene ORF cDNA clone expression plasmid, N-HA tag on other vectors Human HIST1H2BH Gene ORF cDNA clone ... expression plasmid, C-GFPSpark tag. HG15799-ACG. $. 225. Human HIST1H2BH Gene ORF cDNA clone expression plasmid, C-OFPSpark tag ...
Three nuclear genes suppress a yeast mitochondrial splice defect when present in high copy number | SpringerLinkA gene bank of a yeast wild type DNA in the high copy number vector YEp 13 was screened for recombinant plasmids which suppress ... Restriction mapping and cross-hybridization of the inserts revealed that these 17 plasmids contain three different inserts, all ... A total of 17 recombinant plasmids with similar suppressor activity were found. ...
Investigation of diversity of plasmids carrying the blaTEM-52 gene - DTU OrbitSeven out of 10 blaTEM-52 plasmids tested in conjugation experiments were shown to be capable of self-transfer to a plasmid- ... Detected IncI1 plasmids underwent further plasmid multilocus sequence typing. RESULTS: RFLP profiles demonstrated dissemination ... Investigation of diversity of plasmids carrying the blaTEM-52 gene. Publication: Research - peer-review › Journal article - ... Some of the blaTEM-52 plasmids carry replicons that differ from the classical ones. Two novel replicons were detected, IncX1A ...
GenElute Plasmid DNA Miniprep Kit - 350 Purifications | Sigma-AldrichBuy and find information on the GenElute Plasmid Miniprep Kit with 350 purifications great for plasmid DNA isolation from E. ... GenElute™ Plasmid Miniprep Kit sufficient for 350 purifications Synonym: Gen Elute, GenElute™ Plasmid Kit ... Purified plasmid DNA in less than 30 minutes • Purify up to 15 μg of plasmid DNA • No detectable genomic DNA or RNA ... The GenElute Plasmid Miniprep Kit offers a simple, rapid, and cost-effective method for isolating plasmid DNA from recombinant ...
ASMscience | Color PlatesChapter 19) Cloning by gap repair into a linear plasmid. A linear plasmid with "ends-in" homology is generated by PCR with ... Chapter 19) Cloning by gap repair into a linear plasmid. A linear plasmid with "ends-in" homology is generated by PCR with ... a gene or region can be rescued directly from the chromosome onto a plasmid (b). Recombineering restores a circular plasmid ... a gene or region can be rescued directly from the chromosome onto a plasmid (b). Recombineering restores a circular plasmid ...
co-expression of 2 plasmids in E.coli - Molecular Biology - BioForumEach plasmid will carry different gene of interest of course, with 2 different promoters. Also the 2 plasmids have different ... I want to transform 2 plasmids in E.coli in order to co-express 2 different proteins. ... co-expression of 2 plasmids in E.coli - posted in Molecular Biology: Hi everyone!, I would like to describe you briefly the ... Each plasmid will carry different gene of interest of course, with 2 different promoters. Also the 2 plasmids have different ...
Disappearing plasmidsBluescript or low copy plasmids with the same size inserts. When we ,screen our minipreps, we have high amounts of plasmid DNA ... Disappearing plasmids. Arthur Wohlwill U55850 at uic.edu Sat Aug 31 15:12:12 EST 1996 *Previous message: Qiagen refuses to tell ... Disappearing plasmids ,Date: 2 Sep 1996 03:27:23 GMT ,Off and on our laboratory has experienced an annoying problem only with , ... most or all the plasmid. This happens even though we have streaked for ,single colony isolation. Why is this happening? We ...
Human ENSA / Endosulfine alpha Gene ORF cDNA clone expression plasmid, N-Myc tag | SinoBiological... expression plasmid, mutiple tags. Optimized for high expression in mammalian cells. Save up to 60%. ... Each tube contains lyophilized plasmid.. Storage. :. The lyophilized plasmid can be stored at room temperature for three months ... Human ENSA / Endosulfine alpha Gene ORF cDNA clone expression plasmid, N-Myc tag on other vectors Human ENSA / Endosulfine ... Human ENSA / Endosulfine alpha Gene ORF cDNA clone expression plasmid, C-OFPSpark tag. HG14321-ACR. $. 225. ...
Rat IL7/IL-7/Interleukin-7 Gene ORF cDNA clone expression plasmid, C-HA tag | SinoBiological... expression plasmid, mutiple tags. Optimized for high expression in mammalian cells. Save up to 60%. ... Rat SORD Gene ORF cDNA clone expression plasmid, N-OFPSpark tag. *Human PACRG Gene ORF cDNA clone expression plasmid, N-Myc tag ... Each tube contains lyophilized plasmid.. Storage. :. The lyophilized plasmid can be stored at room temperature for three months ... Mouse TXNDC12 Gene ORF cDNA clone expression plasmid, C-HA tag. *Mouse RBM28 Gene ORF cDNA clone expression plasmid, N-OFPSpark ...
Using pBABE plasmids for regular transfection - Molecular CloningUsing pBABE plasmids for regular transfection - (Apr/10/2014 ). Im confused regarding the promoter on the pBABE puro plasmids. ...
マウス BLZF1 Gene ORF cDNA clone expression plasmid, N-Myc タグ | SinoBiological... expression plasmid, mutiple tags. Optimized for high expression in mammalian cells. Save up to 60%. ... Mouse BLZF1 Gene cDNA clone plasmid. MG52016-G. JPY. 18140. マウス BLZF1 Gene ORF cDNA clone expression plasmid, N-Flag タグ. ... マウス BLZF1 Gene ORF cDNA clone expression plasmid, N-Myc タグ on other vectors マウス BLZF1 Gene ORF cDNA clone expression plasmid, C ... マウス BLZF1 Gene ORF cDNA clone expression plasmid, C-OFPSpark タグ. MG52016-ACR. JPY. 54410. ...
Rat RPS6 Gene ORF cDNA clone expression plasmid, N-Myc tag | SinoBiological... expression plasmid, mutiple tags. Optimized for high expression in mammalian cells. Save up to 60%. ... Rat RPS6 Gene ORF cDNA clone expression plasmid, N-Myc tag on other vectors Rat RPS6 Gene ORF cDNA clone expression plasmid, C- ... Each tube contains lyophilized plasmid.. Storage. :. The lyophilized plasmid can be stored at room temperature for three months ... Rat RPS6 Gene ORF cDNA clone expression plasmid, C-OFPSpark tag. RG80730-ACR. $. 225. ...
cloned plasmid expression less than original plasmid - Molecular BiologyYou should be transfecting the same number of molecules of plasmid A vs. plasmid B, which will vary unless the sizes of plasmid ... Since your plasmid with insert is larger than the plasmid without, it will require more plasmid mass to yield the same number ... cloned plasmid expression less than original plasmid - after transfection (Jun/23/2010 ). Hi all,. I have cloned a gene into a ... 150 ng/well of plasmid A + 150 ng/well naked vector (total 300 ng/well). vs.. 200 ng/well of plasmid B (equimolar amount) + 100 ...
GenElute HP Plasmid DNA Maxiprep Kit - 25 Purifications | Sigma-AldrichBuy and find information on the GenElute HP plasmid DNA maxiprep kit with 25 purifications great for plasmid purification at ... GenElute™ HP Plasmid Maxiprep Kit sufficient for 25 purifications Synonym: GenElute™ HP Plasmid Maxiprep Kit ... GenElute™ HP Plasmid Maxiprep Procedure [VIDEO] The GenElute HP Plasmid Maxiprep kit offers a simple, rapid, and cost-effective ... The GenElute HP Plasmid Maxiprep Kit offers a simple, rapid, and cost effective method for isolating plasmid DNA from ...
PCR of entire plasmid followed by self-ligation for mutagenesis- what issues mig - Molecular BiologyPCR of entire plasmid followed by self-ligation for mutagenesis- what issues mig - (Apr/21/2012 ). I've used the quickchange ... The major problem is that PCR tends not to be very efficient for many plasmids due to the large(ish) size of the products. ... I had read that some people found plasmids that seemed to be derived from pcr products with "truncated" ends possibly due to ... will this method reliably lead to generation of plasmids with expected junction between the two ends of the pcr product? Has ...
AID 431834 - Induction of human recombinant topoisomerase 1-mediated cleavage of 32P-labeled linearized pRYG plasmid DNA at 2.5...Induction of human recombinant topoisomerase 1-mediated cleavage of 32P-labeled linearized pRYG plasmid DNA at 2.5 uM after 30 ...
Sequence of Plasmid P1 and R1I am looking , for any site or paper with the complete sequence and description about , the parent plasmids P1 and R1. I would ... Sequence of Plasmid P1 and R1. x x x at santabanta.com Mon Oct 2 04:17:21 EST 2006 *Previous message: Hairpin template in a PCR ... I presume we are talking about E.coli plasmids or in the case of P1 - bacteriophage P1? Nothing in Genbank? Pubmed? Duncan -- I ... Sequence of Plasmid P1 and R1 Date: Fri, 29 Sep 2006 11:40:01 +0100 Historians believe that in newspost ,mailman. ...
Relationship between plasmid size and amount of lipofectamine needed? - Tissue and Cell CultureOr does the amount of lipofectamine just vary by the plasmid? A large plasmid may need less lipofectamine than a smaller ... Relationship between plasmid size and amount of lipofectamine needed? - (Jul/07/2014 ). Is there a correlation between DNA: ... lipofectamine ratio, plasmid size and transfection efficiency? Is it as simple as a small plasmid needs less lipofectamine and ... In my experience, it is relatively rare to have to go outside the 3:1 used for most plasmids. I don't think there is a ...
Addgene: Plasmids related to Trpv4Addgene is a nonprofit plasmid repository. We archive and distribute high quality plasmids from your colleagues. ... Plasmids containing this gene, or a homologous gene.. <"clearfix">t' data-page-length="" data-b-auto-width="false" > ID. ... You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more ...
Addgene: - - Lab PlasmidsLab Plasmids Create a link to this page The - - Lab has deposited plasmids at Addgene for distribution to the research ... Addgene is a nonprofit plasmid repository. We archive and distribute high quality plasmids from your colleagues. ... You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more ... Addgene is a nonprofit plasmid repository dedicated to improving life science research. ...
Plasmid DNA Isolation | Thermo Fisher ScientificInvitrogen Plasmid Isolation kits offer a range options to eliminate the challenges raised by this level of workflow processing ... They are designed to isolate plasmid DNA at the purity and scale you need and are available in a variety of configurations to ... Multi sample processing is often required to complete both plasmid isolation and subsequent downstream experimentation. ... Plasmid isolation is a basic technique performed in most molecular biology laboratories. ...
Addgene: Cohen Lab VIP plasmidsAdam Cohen Lab: Cohen Lab VIP plasmids Unpublished Plasmids from Article. <"clearfix">t' data-page-length="" data-b-auto-width ... Addgene is a nonprofit plasmid repository. We archive and distribute high quality plasmids from your colleagues. ... You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more ...
DNA Protocols & Applications - QIAGENIt also deals with common plasmid DNA procedures, including how to make and transform competent cells, how to culture and ... handle plasmid-containing cells, and commonly used techniques for analysis of genomic DNA. ... section describes considerations for isolation and quantification of both genomic DNA from different sample sources and plasmid ... Plasmid DNA. Bacterial plasmids are closed circular molecules of double-stranded DNA that range in size from 1 to ,200 kb. They ...
Addgene plasmid requestsThe following plasmids have been submitted to Addgene: pET32a-TRX. pET32a-HD16Q. pET32a-HD25Q. pET32a-HD39Q. pET32a-HD46Q. ... If you need any other plasmids from our lab, please send an e-mail to Pamela Bjorkman - (bjorkman[at]caltech.edu) with: ...
Triparental mating: Triparental mating is a form of Bacterial conjugation where a conjugative plasmid present in one bacterial strain assists the transfer of a mobilizable plasmid present in a second bacterial strain into a third bacterial strain. Plasmids are introduced into bacteria for such purposes as transformation, cloning, or transposon mutagenesis.List of strains of Escherichia coli: Escherichia coli is a well studied bacterium that was first identified by Theodor Escherich, after whom it was later named.Symmetry element: A symmetry element is a point of reference about which symmetry operations can take place. In particular, symmetry elements can be centers of inversion, axes of rotation and mirror planes.Coles PhillipsRestriction fragment: A restriction fragment is a DNA fragment resulting from the cutting of a DNA strand by a restriction enzyme (restriction endonucleases), a process called restriction. Each restriction enzyme is highly specific, recognising a particular short DNA sequence, or restriction site, and cutting both DNA strands at specific points within this site.Avery–MacLeod–McCarty experimentLigation-independent cloning: Ligation-independent cloning (LIC) is a form of molecular cloning that is able to be performed without the use of restriction endonucleases or DNA ligase. This allows genes that have restriction sites to be cloned without worry of chopping up the insert.Composite transposon: A composite transposon is similar in function to simple transposons and Insertion Sequence (IS) elements in that it has protein coding DNA segments flanked by inverted, repeated sequences that can be recognized by transposase enzymes. A composite transposon, however, is flanked by two separate IS elements which may or may not be exact replicas.Circular bacterial chromosome: A circular bacterial chromosome is a bacterial chromosome in the form of a molecule of circular DNA. Unlike the linear DNA of most eukaryotes, typical bacterial chromosomes are circular.DNA re-replication: DNA re-replication (or simply rereplication) is an undesirable and possibly fatal occurrence in eukaryotic cells in which the genome is replicated more than once per cell cycle. Rereplication is believed to lead to genomic instability and has been implicated in the pathologies of a variety of human cancers.Ferric uptake regulator family: In molecular biology, the ferric uptake regulator (FUR) family of proteins includes metal ion uptake regulator proteins. These are responsible for controlling the intracellular concentration of iron in many bacteria.Multiple cloning site: A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites - a standard feature of engineered plasmids. Restriction sites within an MCS are typically unique, occurring only once within a given plasmid.DNA sequencer: A DNA sequencer is a scientific instrument used to automate the DNA sequencing process. Given a sample of DNA, a DNA sequencer is used to determine the order of the four bases: G (guanine), C (cytosine), A (adenine) and T (thymine).Recombination (cosmology): In cosmology, recombination refers to the epoch at which charged electrons and protons first became bound to form electrically neutral hydrogen atoms.Note that the term recombination is a misnomer, considering that it represents the first time that electrically neutral hydrogen formed.CccDNA: cccDNA (covalently closed circular DNA) is a special DNA structure that arises during the propagation of some viruses in the cell nucleus and may remain permanently there. It is a double-stranded DNA that originates in a linear form that is ligated by means of DNA ligase to a covalently closed ring.Horizontal gene transfer in evolutionOrigin of replication: The origin of replication (also called the replication origin) is a particular sequence in a genome at which replication is initiated.Technical Glossary Edward K.Silent mutation: Silent mutations are mutations in DNA that do not significantly alter the phenotype of the organism in which they occur. Silent mutations can occur in non-coding regions (outside of genes or within introns), or they may occur within exons.Beta-lactamaseRNA transfection: RNA transfection is the process of deliberately introducing RNA into a living cell. RNA can be purified from cells after lysis or synthesized from free nucleotides either chemically, or enzymatically using an RNA polymerase to transcribe a DNA template.Operon: In genetics, an operon is a functioning unit of genomic DNA containing a cluster of genes under the control of a single promoter. The genes are transcribed together into an mRNA strand and either translated together in the cytoplasm, or undergo trans-splicing to create monocistronic mRNAs that are translated separately, i.Protein primary structure: The primary structure of a peptide or protein is the linear sequence of its amino acid structural units, and partly comprises its overall biomolecular structure. By convention, the primary structure of a protein is reported starting from the amino-terminal (N) end to the carboxyl-terminal (C) end.BacitracinGC box: In molecular biology, a GC box is a distinct pattern of nucleotides found in the promoter region of some eukaryotic genes upstream of the TATA box and approximately 110 bases upstream from the transcription initiation site. It has a consensus sequence GGGCGG which is position dependent and orientation independent.Eukaryotic transcription: Eukaryotic transcription is the elaborate process that eukaryotic cells use to copy genetic information stored in DNA into units of RNA replica. Gene transcription occurs in both eukaryotic and prokaryotic cells.RhizobiaEscherichia coli (molecular biology): Escherichia coli (; commonly abbreviated E. coli) is a gammaproteobacterium commonly found in the lower intestine of warm-blooded organisms (endotherms).Colicin: A colicin is a type of bacteriocin produced by and toxic to some strains of Escherichia coli.* Feldgarden, M.PBR322: pBR322 is a plasmid and was one of the first widely used E. coli cloning vectors.Opine: Opine biosynthesis is catalyzed by specific enzymes encoded by genes contained in a small segment of DNA (known as the T-DNA, for 'transfer DNA'), which is part of the Ti plasmid, inserted by the bacterium into the plant genome. The opines are used by the bacterium as an important source of nitrogen and energy.Virulence: Virulence is, by MeSH definition, the degree of pathogenicity within a group or species of parasites as indicated by case fatality rates and/or the ability of the organism to invade the tissues of the host. The pathogenicity of an organism - its ability to cause disease - is determined by its virulence factors.Thermal cyclerNaked DNA: Naked DNA is histone-free DNA that is passed from cell to cell during a gene transfer process called transformation or transfection. In transformation, purified or naked DNA is taken up by the recipient cell which will give the recipient cell a new characteristic or phenotype.Chromosome regionsGene electrotransfer: Gene electrotransfer is a versatile biotechnology technique that enables the transfer of genetic material into prokaryotic or eukaryotic cells. It is based on a physical method named electroporation, where a transient increase in the permeability of cell membrane is achieved when submitted to short and intense electric pulses, thus enabling the transport of large molecules (naked plasmid DNA, antisense oligonucleotides, siRNA) into cells that otherwise cannot permeate through the cell membrane.Bismuth sulfite agar: Bismuth sulfite agar is a type of agar media used to isolate Salmonella species. It uses glucose as a primary source of carbon.Pseudomonas alkanolytica: Pseudomonas alkanolytica is a Gram-negative soil bacterium that produces Coenzyme A. Because this organism is patented,Nakao Y, Kuno M.StreptomycinKlebsiella pneumoniaOpen reading frame: In molecular genetics, an open reading frame (ORF) is the part of a reading frame that has the potential to code for a protein or peptide. An ORF is a continuous stretch of codons that do not contain a stop codon (usually UAA, UAG or UGA).Streptomyces kanamyceticus: Streptomyces kanamyceticus is a bacterial species in the genus Streptomyces. It is the species from which the antibiotic kanamycin is isolated.DNA condensation: DNA condensation refers to the process of compacting DNA molecules in vitro or in vivo. Mechanistic details of DNA packing are essential for its functioning in the process of gene regulation in living systems.Direct repeat: Direct repeats are a type of genetic sequence that consists of two or more repeats of a specific sequence.Resistome: The resistome is a proposed expression by Gerard D. Wright for the collection of all the antibiotic resistance genes and their precursors in both pathogenic and non-pathogenic bacteria.Global microbial identifier: The genomic epidemiological database for global identification of microorganisms or global microbial identifier (GMI) is a platform for storing whole genome sequencing (WGS) data of microorganisms, for the identification of relevant genes and for the comparison of genomes to detect and track-and-trace infectious disease outbreaks and emerging pathogens. The database holds two types of information: 1) genomic information of microorganisms, linked to, 2) metadata of those microorganism such as epidemiological details.Chloramphenicol acetyltransferasePulsenet: PulseNet is a network run by the Centers for Disease Control and Prevention (CDC) which brings together public health and food regulatory agency laboratories around the United States.http://www.Nucleic acid structure: Nucleic acid structure refers to the structure of nucleic acids such as DNA and RNA. Chemically speaking, DNA and RNA are very similar.DNA-binding proteinPhenotype microarray: The phenotype microarray approach is a technology for high-throughput phenotyping of cells.Branching order of bacterial phyla (Gupta, 2001): There are several models of the Branching order of bacterial phyla, one of these was proposed in 2001 by Gupta based on conserved indels or protein, termed "protein signatures", an alternative approach to molecular phylogeny. Some problematic exceptions and conflicts are present to these conserved indels, however, they are in agreement with several groupings of classes and phyla.Enterococcus faecalis: Enterococcus faecalis – formerly classified as part of the group D Streptococcus system – is a Gram-positive, commensal bacterium inhabiting the gastrointestinal tracts of humans and other mammals. Like other species in the genus Enterococcus, E.Molar mass distribution: In linear polymers the individual polymer chains rarely have exactly the same degree of polymerization and molar mass, and there is always a distribution around an average value. The molar mass distribution (or molecular weight distribution) in a polymer describes the relationship between the number of moles of each polymer species (Ni) and the molar mass (Mi) of that species.CTXφ Bacteriophage: The CTXφ bacteriophage is a filamentous bacteriophage that contains the genetic material needed by the Vibrio cholerae bacterium for the production of cholera toxin, or CT. CTXφ is a positive virus with single-stranded DNA (ssDNA).AmpicillinRestriction site: Restriction sites, or restriction recognition sites, are locations on a DNA molecule containing specific (4-8 base pairs in length) sequences of nucleotides, which are recognized by restriction enzymes. These are generally palindromic sequences (because restriction enzymes usually bind as homodimers), and a particular restriction enzyme may cut the sequence between two nucleotides within its recognition site, or somewhere nearby.Streptomyces peucetius: Streptomyces peucetius ATCC 27952Homing endonuclease: The homing endonucleases are a collection of endonucleases encoded either as freestanding genes within introns, as fusions with host proteins, or as self-splicing inteins. They catalyze the hydrolysis of genomic DNA within the cells that synthesize them, but do so at very few, or even singular, locations.Chromosome engineering: Chromosome engineering is "the controlled generation of chromosomal deletions, inversions, or translocations with defined endpoints." For: By combining chromosomal translocation, chromosomal inversion,and chromosomal deletion, chromosome engineering has been shown to identify the underlying genes that cause certain diseases in mice.Trimethoprim
(1/43669) Stabilization of poly-L-lysine/DNA polyplexes for in vivo gene delivery to the liver.
We are developing a self-assembling non-viral in vivo gene delivery vehicle based on poly-l-lysine and plasmid DNA. We have characterized poly-l-lysines of different chain lengths for DNA condensation and strength of DNA binding. Poly-l-lysine chains >20 residues bound DNA efficiently in physiological saline, while shorter chains did not. Attachment of asialoorosomucoid to PLL increased the PLL chain length required for efficient DNA binding in saline and for efficient DNA condensation. By electron microscopy, poly-l-lysine/DNA polyplexes appeared as toroids 25-50 nm in diameter or rods 40-80 nm long; conjugation of asialoorosomucoid to the polylysine component increased the size of resulting polyplexes to 50-90 nm. In water, poly-l-lysine and asialoorosomucoid-PLL polyplexes have effective diameters of 46 and 87.6 nm, respectively. Polyplexes containing only poly-l-lysine and DNA aggregated in physiological saline at all charge ratios and aggregated at neutral charge ratios in water. Attachment of asialoorosomucoid lessened, but did not eliminate, the aggregation of PLL polyplexes, and did not result in efficient delivery of polyplexes to hepatocytes. Conjugation of polyethylene glycol to poly-l-lysine sterically stabilized resulting polyplexes at neutral charge ratios by shielding the surfaces. For efficient in vivo gene delivery, polyplexes will need to be sterically stabilized to prevent aggregation and interaction with serum components. (+info)
(2/43669) Cloning and characterisation of a novel ompB operon from Vibrio cholerae 569B.
The ompB operon of Vibrio cholerae 569B has been cloned and fully sequenced. The operon encodes two proteins, OmpR and EnvZ, which share sequence identity with the OmpR and EnvZ proteins of a variety of other bacteria. Although the order of the ompR and envZ genes of V. cholerae is similar to that of the ompB operon of E. coli, S. typhimurium and X. nematophilus, the Vibrio operon exhibits a number of novel features. The structural organisation and features of the V. cholerae ompB operon are described. (+info)
(3/43669) Co-expression of glutathione S-transferase with methionine aminopeptidase: a system of producing enriched N-terminal processed proteins in Escherichia coli.
We describe here an Escherichia coli expression system that produces recombinant proteins enriched in the N-terminal processed form, by using glutathione S-transferase cGSTM1-1 and rGSTT1-1 as models, where c and r refer to chick and rat respectively. Approximately 90% of the cGSTM1-1 or rGSTT1-1 overexpressed in E. coli under the control of a phoA promoter retained the initiator methionine residue that was absent from the mature isoenzymes isolated from tissues. The amount of initiator methionine was decreased to 40% of the expressed cGSTM1-1 when the isoenzyme was co-expressed with an exogenous methionine aminopeptidase gene under the control of a separate phoA promoter. The recombinant proteins expressed were mainly methionine aminopeptidase. The yield of cGSTM1-1 was decreased to 10% of that expressed in the absence of the exogenous methionine aminopeptidase gene. By replacing the phoA with its natural promoter, the expression of methionine aminopeptidase decreased drastically. The yield of the co-expressed cGSTM1-1 was approx. 60% of that in the absence of the exogenous methionine aminopeptidase gene; approx. 65% of the initiator methionine residues were removed from the enzyme. Under similar conditions, N-terminal processing was observed in approx. 70% of the recombinant rGSTT1-1 expressed. By increasing the concentration of phosphate in the growth medium, the amount of initiator methionine on cGSTM1-1 was decreased to 14% of the overexpressed isoenzymes, whereas no further improvement could be observed for rGSTT1-1. The initiator methionine residue does not affect the enzymic activities of either cGSTM1-1 or rGSTT1-1. However, the epoxidase activity and the 4-nitrobenzyl chloride-conjugating activity of the purified recombinant rGSTT1-1 are markedly higher that those reported recently for the same isoenzyme isolated from rat livers. (+info)
(4/43669) Expression of the plague plasminogen activator in Yersinia pseudotuberculosis and Escherichia coli.
Enteropathogenic yersiniae (Yersinia pseudotuberculosis and Yersinia enterocolitica) typically cause chronic disease as opposed to the closely related Yersinia pestis, the causative agent of bubonic plague. It is established that this difference reflects, in part, carriage by Y. pestis of a unique 9.6-kb pesticin or Pst plasmid (pPCP) encoding plasminogen activator (Pla) rather than distinctions between shared approximately 70-kb low-calcium-response, or Lcr, plasmids (pCD in Y. pestis and pYV in enteropathogenic yersiniae) encoding cytotoxic Yops and anti-inflammatory V antigen. Pla is known to exist as a combination of 32.6-kDa (alpha-Pla) and slightly smaller (beta-Pla) outer membrane proteins, of which at least one promotes bacterial dissemination in vivo and degradation of Yops in vitro. We show here that only alpha-Pla accumulates in Escherichia coli LE392/pPCP1 cultivated in enriched medium and that either autolysis or extraction of this isolate with 1.0 M NaCl results in release of soluble alpha and beta forms possessing biological activity. This process also converted cell-bound alpha-Pla to beta-Pla and smaller forms in Y. pestis KIM/pPCP1 and Y. pseudotuberculosis PB1/+/pPCP1 but did not promote solubilization. Pla-mediated posttranslational hydrolysis of pulse-labeled Yops in Y. pseudotuberculosis PB1/+/pPCP1 occurred more slowly than that in Y. pestis but was otherwise similar except for accumulation of stable degradation products of YadA, a pYV-mediated fibrillar adhesin not encoded in frame by pCD. Carriage of pPCP by Y. pseudotuberculosis did not significantly influence virulence in mice. (+info)
(5/43669) The virulence plasmid-encoded impCAB operon enhances survival and induced mutagenesis in Shigella flexneri after exposure to UV radiation.
Upon exposure to UV radiation, Shigella flexneri SA100 displayed survival and mutation frequencies comparable to those of Escherichia coli AB1157, which contains a functional UmuDC error-prone DNA repair system. Survival of SA100 after UV irradiation was associated with the presence of the 220-kb virulence plasmid, pVP. This plasmid encodes homologues of ImpA and ImpB, which comprise an error-prone DNA repair system encoded on plasmid TP110 that was initially identified in Salmonella typhimurium, and ImpC, encoded upstream of ImpA and ImpB. Although the impB gene was present in representatives of all four species of Shigella, not all isolates tested contained the gene. Shigella isolates that lacked impB were more sensitive to UV radiation than isolates that contained impB. The nucleotide sequence of a 2.4-kb DNA fragment containing the imp operon from S. flexneri SA100 pVP was 96% identical to the imp operon from the plasmid TP110. An SA100 derivative with a mutation in the impB gene had reduced survival following UV irradiation and less UV-induced mutagenesis relative to the parental strain. We also found that S. flexneri contained a chromosomally encoded umuDC operon; however, the umuDC promoter was not induced by exposure to UV radiation. This suggests that the imp operon but not the umuDC operon contributes to survival and induced mutagenesis in S. flexneri following exposure to UV radiation. (+info)
(6/43669) Molecular and evolutionary analysis of Borrelia burgdorferi 297 circular plasmid-encoded lipoproteins with OspE- and OspF-like leader peptides.
We previously described two OspE and three OspF homologs in Borrelia burgdorferi 297 (D. R. Akins, S. F. Porcella, T. G. Popova, D. Shevchenko, S. I. Baker, M. Li, M. V. Norgard, and J. D. Radolf, Mol. Microbiol. 18:507-520, 1995; D. R. Akins, K. W. Bourell, M. J. Caimano, M. V. Norgard, and J. D. Radolf, J. Clin. Investig. 101:2240-2250, 1998). In this study, we characterized four additional lipoproteins with OspE/F-like leader peptides (Elps) and demonstrated that all are encoded on plasmids homologous to cp32 and cp18 from the B31 and N40 strains, respectively. Statistical analysis of sequence similarities using the binary comparison algorithm revealed that the nine lipoproteins from strain 297, as well as the OspE, OspF, and Erp proteins from the N40 and B31 strains, fall into three distinct families. Based upon the observation that these lipoproteins all contain highly conserved leader peptides, we now propose that the ancestors of each of the three families arose from gene fusion events which joined a common N terminus to unrelated proteins. Additionally, further sequence analysis of the strain 297 circular plasmids revealed that rearrangements appear to have played an important role in generating sequence diversity among the members of these three families and that recombinational events in the downstream flanking regions appear to have occurred independently of those within the lipoprotein-encoding genes. The association of hypervariable regions with genes which are differentially expressed and/or subject to immunological pressures suggests that the Lyme disease spirochete has exploited recombinatorial processes to foster its parasitic strategy and enhance its immunoevasiveness. (+info)
(7/43669) Reverse transcription-nested polymerase chain reaction for detecting p40 RNA of Borna disease virus, without risk of plasmid contamination.
Several methods for the detection of Borna disease virus (BDV) RNA have been reported, one being the reverse transcription-nested polymerase chain reaction (RT-nested PCR) method. However, due to the possibility of contamination of the cloned DNA in a reaction tube, false-positive results might be obtained by RT-nested PCR. To detect only BDV RNA without anxiety of contamination, we developed an RT-nested PCR system using "mRNA selective PCR kit". Using this system, cDNA of BDV p40 in the plasmid (up to 5 x 10(7) molecules) was not amplified. BDV specific sequence was amplified from total RNA (more than 50 pg) of MDCK/BDV cells, which were persistently infected with BDV. These results indicate that this mRNA selective RT-nested PCR system can specifically amplify target RNA as distinguished from plasmid contaminated. (+info)
(8/43669) Diversity of rhizobia associated with Amorpha fruticosa isolated from Chinese soils and description of Mesorhizobium amorphae sp. nov.
Fifty-five Chinese isolates from nodules of Amorpha fruticosa were characterized and compared with the type strains of the species and genera of bacteria which form nitrogen-fixing symbioses with leguminous host plants. A polyphasic approach, which included RFLP of PCR-amplified 16S rRNA genes, multilocus enzyme electrophoresis (MLEE), DNA-DNA hybridization, 16S rRNA gene sequencing, electrophoretic plasmid profiles, cross-nodulation and a phenotypic study, was used in the comparative analysis. The isolates originated from several different sites in China and they varied in their phenotypic and genetic characteristics. The majority of the isolates had moderate to slow growth rates, produced acid on YMA and harboured a 930 kb symbiotic plasmid (pSym). Five different RFLP patterns were identified among the 16S rRNA genes of all the isolates. Isolates grouped by PCR-RFLP of the 16S rRNA genes were also separated into groups by variation in MLEE profiles and by DNA-DNA hybridization. A representative isolate from each of these DNA homology groups had a separate position in a phylogenetic tree as determined from sequencing analysis of the 16S rRNA genes. A new species, Mesorhizobium amorphae, is proposed for the majority of the isolates, which belonged to a moderately slow- to slow-growing, acid-producing group based upon their distinct phylogenetic position, their unique electrophoretic type, their low DNA homology with reference strains representing the species within the genus Mesorhizobium and their distinct phenotypic features. Strain ACCC 19665 was chosen as the type strain for M. amorphae sp. nov. (+info)
- A large plasmid may need less lipofectamine than a smaller plasmid or vice versa. (protocol-online.org)
- Use a plasmid vector to transform bacteria with genes for Green Fluorescent Protein (GFP) and antibiotic resistance in a controlled experiment. (vernier.com)
- One technique to correct genetic defects in bacteria is to introduce the corrected gene back into bacteria utilising plasmids, a technique known as a-complementation. (astralscientific.com.au)
- High-frequency conjugal plasmid transfer from gram-negative Escherichia coli to various gram-positive coryneform bacteria. (uni-bielefeld.de)
- Schäfer A, Kalinowski J, Simon R, Seep-Feldhaus AH, Pühler A. High-frequency conjugal plasmid transfer from gram-negative Escherichia coli to various gram-positive coryneform bacteria. (uni-bielefeld.de)
- Frame 6 shows the membrane proteins encoded by the tra genes of the conjugative plasmid of the donor bacterium fusing the membranes of the two bacteria, creating an opening between them. (asmscience.org)
- OBJECTIVES: To investigate the diversity of plasmids that carry blaTEM-52 genes among Escherichia coli and Salmonella enterica originating from animals, meat products and humans. (dtu.dk)
- Bio-Rad's pGLO plasmid incorporates the arabinose promoter, but the genes involved in the breakdown of arabinose have been replaced with the jellyfish gene encoding GFP. (bio-rad.com)
- A conjugative plasmid is self-transmissible, in that it possesses all the necessary genes for that plasmid to transmit itself to another bacterium by conjugation. (asmscience.org)
- Conjugation genes known as tra genes enable the bacterium to form a mating pair with another organism, while oriT (origin of transfer) sequences determine where on the plasmid DNA transfer is initiated by serving as the replication start site where DNA replication enzymes will nick the DNA to initiate DNA replication and transfer. (asmscience.org)
- In addition, mobilizable plasmids that lack the tra genes for self-transmissibility but possess the oriT for initiation of DNA transfer may also be transferred by conjugation if the bacterium containing them also possesses a conjugative plasmid. (asmscience.org)
- The tra genes of the conjugative plasmid enable a mating pair to form and the oriT of the mobilizable plasmid enable the DNA to replicate as it moves through the conjugative bridge. (asmscience.org)
- METHODS: A collection of 22 blaTEM-52-encoding plasmids was characterized by restriction fragment length polymorphism (RFLP), replicon typing (by PCR or replicon sequencing), susceptibility testing, assessment of plasmid ability to self-transfer by conjugation and typing of the genetic environment of the blaTEM-52 gene. (dtu.dk)
- Seven out of 10 blaTEM-52 plasmids tested in conjugation experiments were shown to be capable of self-transfer to a plasmid-free E. coli recipient. (dtu.dk)
- This animation shows bacterial conjugation resulting in the transfer of a conjugative plasmid and a mobilizable plasmid. (asmscience.org)
- Conjugation involves a donor bacterium that contains a conjugative plasmid and a recipient cell that does not. (asmscience.org)
- We report on the mobilization of shuttle plasmids from gram-negative Escherichia coli to gram-positive corynebacteria mediated by P-type transfer functions. (uni-bielefeld.de)
- Introduction of plasmids into corynebacteria was markedly enhanced after heat treatment of the recipient cells. (uni-bielefeld.de)
- High-frequency plasmid transfer was also observed when the restriction system of the recipient was mutated. (uni-bielefeld.de)
- Frames 4 and 5 show the donor bacterium (with chromosome and conjugative plasmid) binding the recipient bacterium that contains a chromosome but lacks the conjugative plasmid. (asmscience.org)
- Frames 7, 8, 9 and 10 show the conjugative plasmid being nicked at the oriT by nuclease products of the tra gene, this nuclease also has helicase activity and will unwind the strand that is now being transferred to the recipient bacterium. (asmscience.org)
- Bio-Rad's exclusive pGLO plasmid is constructed with the jellyfish gene that encodes green fluorescent protein (GFP), an antibiotic-resistance gene that encodes β-lactamase protein, and the araC gene encoding a regulator protein that turns the GFP gene on and off. (bio-rad.com)
- 5 µg) of single copy plasmids (size=12.9 Kbp)? (protocol-online.org)
- Relationship between plasmid size and amount of lipofectamine needed? (protocol-online.org)
- Is there a correlation between DNA:lipofectamine ratio, plasmid size and transfection efficiency? (protocol-online.org)
- I feel that it is more complex and needs to be determined empirically or the reagent protocols would specify plasmid size. (protocol-online.org)
- I don't think there is a relationship between plasmid size and amount of lipofectamine, but perhaps there is something related to the overall charge on the plasmid. (protocol-online.org)
- RESULTS: RFLP profiles demonstrated dissemination of blaTEM-52 in Denmark (imported meat from Germany), France, Belgium and the Netherlands from 2000 to 2006 by mainly two different plasmids, one encoding blaTEM-52b (IncX1A, 45 kb) and the other blaTEM-52c (IncI1, 80 kb). (dtu.dk)
- In addition, blaTEM-52b was also found to be located on various other plasmids belonging to IncA/C and IncL/M, while blaTEM-52c was found on IncN-like as well as on IncR plasmids. (dtu.dk)