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.
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.
A class of plasmids that transfer antibiotic resistance from one bacterium to another by conjugation.
Plasmids encoding bacterial exotoxins (BACTERIOCINS).
Deoxyribonucleic acid that makes up the genetic material of bacteria.
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
The functional hereditary units of BACTERIA.
Vertical transmission of hereditary characters by DNA from cytoplasmic organelles such as MITOCHONDRIA; CHLOROPLASTS; and PLASTIDS, or from PLASMIDS or viral episomal DNA.
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.
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.
The heritable modification of the properties of a competent bacterium by naked DNA from another source. The uptake of naked DNA is a naturally occuring phenomenon in some bacteria. It is often used as a GENE TRANSFER TECHNIQUE.
The 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.
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.
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)
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.
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.
Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.
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).
The process by which a DNA molecule is duplicated.
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.
Proteins found in any species of bacterium.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
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)
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.
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
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.
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.
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)
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).
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 in which agar or agarose gel is used as the diffusion medium.
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.
A naphthacene antibiotic that inhibits AMINO ACYL TRNA binding during protein synthesis.
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.
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.
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.
In bacteria, a group of metabolically related genes, with a common promoter, whose transcription into a single polycistronic MESSENGER RNA is under the control of an OPERATOR REGION.
The 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.
Substances that reduce the growth or reproduction of BACTERIA.
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.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
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.
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.
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.
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.
Proteins obtained from ESCHERICHIA COLI.
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.
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.
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.
Nonsusceptibility of bacteria to the action of TETRACYCLINE which inhibits aminoacyl-tRNA binding to the 30S ribosomal subunit during protein synthesis.
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)
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.
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.
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.
Any method used for determining the location of and relative distances between genes on a chromosome.
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.
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.
A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in nature. Some species are pathogenic for humans, animals, and plants.
An antibiotic produced by the soil actinomycete Streptomyces griseus. It acts by inhibiting the initiation and elongation processes during protein synthesis.
Gram-negative, non-motile, capsulated, gas-producing rods found widely in nature and associated with urinary and respiratory infections in humans.
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).
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.
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).
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).
Established cell cultures that have the potential to propagate indefinitely.
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.
A beta-lactamase preferentially cleaving penicillins. (Dorland, 28th ed) EC 3.5.2.-.
Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria.
Deoxyribonucleic acid that makes up the genetic material of viruses.
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).
The genetic complement of a BACTERIA as represented in its DNA.
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).
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)
Plasmids which determine the ability of a bacterium to ferment lactose.
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.
A serotype of Salmonella enterica that is a frequent agent of Salmonella gastroenteritis in humans. It also causes PARATYPHOID FEVER.
The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.
Genes which regulate or circumscribe the activity of other genes; specifically, genes which code for PROTEINS or RNAs which have GENE EXPRESSION REGULATION functions.
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.
Any tests that demonstrate the relative efficacy of different chemotherapeutic agents against specific microorganisms (i.e., bacteria, fungi, viruses).
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.
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.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
Deoxyribonucleic acid that makes up the genetic material of fungi.
The sequential location of genes on a chromosome.
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
The relationships of groups of organisms as reflected by their genetic makeup.
A species of gram-positive, coccoid bacteria commonly isolated from clinical specimens and the human intestinal tract. Most strains are nonhemolytic.
A complex of antibiotic substances produced by Streptomyces tenebrarius.
The sum of the weight of all the atoms in a molecule.
Nonsusceptibility of bacteria to the antibiotic KANAMYCIN, which can bind to their 70S ribosomes and cause misreading of messenger RNA.
Viruses whose hosts are bacterial cells.
Infections with bacteria of the species ESCHERICHIA COLI.
Semi-synthetic derivative of penicillin that functions as an orally active broad-spectrum antibiotic.
One of the Type II site-specific deoxyribonucleases (EC 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.-.
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.
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
Directed modification of the gene complement of a living organism by such techniques as altering the DNA, substituting genetic material by means of a virus, transplanting whole nuclei, transplanting cell hybrids, etc.
A 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.
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.
The functional hereditary units of VIRUSES.
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.
Nonsusceptibility of bacteria to the action of TRIMETHOPRIM.
DNA elements that include the component genes and insertion site for a site-specific recombination system that enables them to capture mobile gene cassettes.
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 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.
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.
The type species of gram-negative bacteria in the genus SPIROPLASMA, family SPIROPLASMATACEAE, causing citrus stubborn disease.
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.
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.
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.
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 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 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.
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.
Proteins found in any species of virus.
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 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.
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.
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
A species of gram-positive bacteria that is a common soil and water saprophyte.
Viruses whose host is Escherichia coli.
Nonsusceptibility of an organism to the action of penicillins.
A non-pathogenic species of LACTOCOCCUS found in DAIRY PRODUCTS and responsible for the souring of MILK and the production of LACTIC ACID.
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.
Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.
A lactose-fermenting bacterium causing dysentery.
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.
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.
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.
Actual loss of portion of a chromosome.
One of the Type II site-specific deoxyribonucleases (EC 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.-.
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.
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.
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.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
A methylsulfonyl analog of CHLORAMPHENICOL. It is an antibiotic and immunosuppressive agent.
Infections with bacteria of the family ENTEROBACTERIACEAE.
Infections with bacteria of the genus KLEBSIELLA.
Plasmids controlling the synthesis of hemolysin by bacteria.
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.
The most common etiologic agent of GAS GANGRENE. It is differentiable into several distinct types based on the distribution of twelve different toxins.
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.
Nonsusceptibility of a microbe to the action of ampicillin, a penicillin derivative that interferes with cell wall synthesis.
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)
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.
A species of gram-negative bacteria causing URINARY TRACT INFECTIONS and SEPTICEMIA.
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.
The functional hereditary units of FUNGI.
Process of determining and distinguishing species of bacteria or viruses based on antigens they share.

Stabilization of poly-L-lysine/DNA polyplexes for in vivo gene delivery to the liver. (1/43669)

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)

Cloning and characterisation of a novel ompB operon from Vibrio cholerae 569B. (2/43669)

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)

Co-expression of glutathione S-transferase with methionine aminopeptidase: a system of producing enriched N-terminal processed proteins in Escherichia coli. (3/43669)

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)

Expression of the plague plasminogen activator in Yersinia pseudotuberculosis and Escherichia coli. (4/43669)

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)

The virulence plasmid-encoded impCAB operon enhances survival and induced mutagenesis in Shigella flexneri after exposure to UV radiation. (5/43669)

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)

Molecular and evolutionary analysis of Borrelia burgdorferi 297 circular plasmid-encoded lipoproteins with OspE- and OspF-like leader peptides. (6/43669)

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)

Reverse transcription-nested polymerase chain reaction for detecting p40 RNA of Borna disease virus, without risk of plasmid contamination. (7/43669)

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)

Diversity of rhizobia associated with Amorpha fruticosa isolated from Chinese soils and description of Mesorhizobium amorphae sp. nov. (8/43669)

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)

Antibiotic resistance represents a significant public health problem. When resistance genes are mobile, being carried on plasmids or phages, their spread can be greatly accelerated. Plasmids in particular have been implicated in the spread of antibiotic resistance genes. However, the selective pressures which favour plasmid-carried resistance genes have not been fully established. Here we address this issue with mathematical models of plasmid dynamics in response to different antibiotic treatment regimes. We show that transmission of plasmids is a key factor influencing plasmid-borne antibiotic resistance, but the dosage and interval between treatments is also important. Our results also hold when plasmids carrying the resistance gene are in competition with other plasmids that do not carry the resistance gene. By altering the interval between antibiotic treatments, and the dosage of antibiotic, we show that different treatment regimes can select for either plasmid-carried, or chromosome-carried,
Individual bacterial cells may contain several different types of plasmids and in some cases more than 10 at a time. Plasmids are generally isolated from the bacterial cells in the supercoiled configuration. So far, thousands of different types of plasmids have been isolated. More than 300 different types of naturally occurring plasmids have been isolated from E.coli alone. Though, plasmids are not considered as part of the cells genome, when a bacterial cell divides each daughter cells receives a copy of each plasmid. Plasmids can also be transferred from one bacterial cell to another by the process called conjugation. Plasmids that govern their own transfer by conjugation are called conjugative plasmids but not all plasmids are conjugative.. ...
Despite a fitness cost imposed on bacterial hosts, large conjugative plasmids play a key role in the diffusion of resistance determinants, such as CTX-M extended-spectrum beta-lactamases. Among the large conjugative plasmids, IncF plasmids are the most predominant group, and an F2A1B- IncF-type plasmid encoding a CTX-M-15 variant was recently described as being strongly associated with the emerging worldwide Escherichia coli sequence type 131 (ST131)-O25bH4 H30Rx/C2 sublineage. In this context, we investigated the fitness cost of narrow-range F-type plasmids, including the F2A1B- IncF-type CTX-M-15 plasmid, and of broad-range C-type plasmids in the K-12-like J53-2 E. coli strain. Although all plasmids imposed a significant fitness cost to the bacterial host immediately after conjugation, we show, using an experimental-evolution approach, that a negative impact on the fitness of the host strain was maintained throughout 1,120 generations with the IncC-IncR plasmid, regardless of the presence or absence
Götting C, Thierbach G, Pühler A, Kalinowski J. Versatile low-copy-number plasmids for temperature-inducible overexpression of bacterial genes in Escherichia coli. BIOTECHNIQUES. 1998;24(3):362 ...
The prevalence of 20-30 kb plasmids, almost half of which belong to only three restriction types by RFLP analysis, in staphylococci isolated from sources very distant in time and space suggests that these nonconjugative plasmids are surprisingly widespread for non-self-mobile plasmids. Plasmids in this size range can potentially be transferred by transducing phages (Lindsay and Holden 2006; Malachowa and Deleo 2010; Smillie et al. 2010); most phage genomes identified in staphylococci are ,40 kb, and transduction is thought to be restricted by phage genome size (Smillie et al. 2010). More of these 20-30 kb plasmids may be mobilizable than is apparent with current genome data if they contain mob genes not yet identified as such. However, the scarcity of conjugative plasmids (only 12 in total) implies that mobilization is rare and that staphylococcal plasmid transfer occurs mainly by transduction (Lindsay and Holden 2004; Lindsay 2010). The now larger dataset makes the mechanism of intercellular ...
The presence of the amplified product PR is oraut by electrophoresis in 1% agarose gel.. Example 2. Construction of recombinant plasmid DNA pFastBac-G2R-IgG.. 5-10 μg of plasmid pFastBac [28] hydrolyzing the restriction endonucleases BamHI and HindIII; 1-5 μg of the amplified product corresponding gene G2R UPE without termination codon, hydrolyzing the restriction endonucleases BamHI and XbaI; the plasmid pBluescript-IgG1 hydrolyzing the restriction endonucleases XbaI and HindIII in standard conditions. The resulting fragments are electrophoresis in 1% agarose gel, followed by elution. 0.2 μg of the vector and 0.6 µg of fragments are ligated under standard conditions and received ligase mixture transform competent cells of E. coli strain XL-blue. Cellular Apr-clones grown at 37°C in LB medium containing 100 μg/ml ampicillin, until the stationary phase. Recombinant plasmid DNA secrete according to standard methods and analyzed by restriction endonucleases BamHI, XbaI and HindIII. Clones ...
Although IncP-1 plasmids are important for horizontal gene transfer among bacteria, in particular antibiotic resistance spread, so far only three plasmids from the subgroup IncP-1 alpha have been completely sequenced. In this study we doubled this number. The three IncP-1 alpha plasmids pB5, pB11 and pSP21 were isolated from bacteria of two different sewage treatment plants and sequenced by a combination of next-generation and capillary sequencing technologies. A comparative analysis including the previously analysed IncP-1 alpha plasmids RK2, pTB11 and pBS228 revealed a highly conserved plasmid backbone (at least 99.9% DNA sequence identity) comprising 54 core genes. The accessory elements of the plasmid pB5 constitute a class 1 integron interrupting the parC gene and an IS6100 copy inserted into the integron. In addition, the tetracycline resistance genes tetAR and the ISTB11-like element are located between the klc operon and the trfA-ssb operon. Plasmid pB11 is loaded with a Tn5053-like ...
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. 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. Detected IncI1 plasmids underwent further plasmid multilocus sequence typing. 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). 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 ...
In this semirural community, we found that numerically dominant commensal E. coli strains (showing similar antimicrobial resistance and same antibiotic resistance genes) colonizing children and domestic animals in the same period of time and in the same community are genotypically diverse. We also found that plasmids carrying the same antibiotic resistance genes were distinct, which is consistent with recent reports showing that AMR genes move frequently among different plasmids (28, 29). Our research suggests that a common pool of AMR genes could be cocirculating on different plasmids among different E. coli clones in a community (Table 2)-probably through dissemination mediated by transposons, integrons, or gene cassettes (28, 30). Even when the same resistance gene alleles and same plasmid replicon types were identified across isolates, the plasmids harboring these traits were still distinct. We also found potential evidence of Tn2 participation in mobility of the gene blaTEM-1B, as we found ...
Plasmids, DNA (or rarely RNA) molecules which replicate in cells autonomously (independently of chromosomes) as non-essential genetic elements, play important roles for microbes grown under specific environmental conditions as well as in scientific laboratories and in biotechnology. For example, bacterial plasmids are excellent models in studies on regulation of DNA replication, and their derivatives are the most commonly used vectors in genetic engineering. Detailed mechanisms of replication initiation, which is the crucial process for efficient maintenance of plasmids in cells, have been elucidated for several plasmids. However, to understand plasmid biology, it is necessary to understand regulation of plasmid DNA replication in response to different environmental conditions in which host cells exist. Knowledge of such regulatory processes is also very important for those who use plasmids as expression vectors to produce large amounts of recombinant proteins. Variable conditions in large-scale
Plasmids are important members of the bacterial mobile gene pool, and are among the most important contributors to horizontal gene transfer between bacteria. They typically harbour a wide spectrum of host beneficial traits, such as antibiotic resistance, inserted into their backbones. Although these inserted elements have drawn considerable interest, evolutionary information about the plasmid backbones, which encode plasmid related traits, is sparse. Here we analyse 25 complete backbone genomes from the broad-host-range IncP-1 plasmid family. Phylogenetic analysis reveals seven clades, in which two plasmids that we isolated from a marine biofilm represent a novel clade. We also found that homologous recombination is a prominent feature of the plasmid backbone evolution. Analysis of genomic signatures indicates that the plasmids have adapted to different host bacterial species. Globally circulating IncP-1 plasmids hence contain mosaic structures of segments derived from several parental plasmids that
A plasmid partition system is a mechanism that assures the stable transmission of plasmids during bacterial cell division. Each plasmid has its independent replication system which controls the number of copies of the plasmid in a cell. The higher the copy number is, the more likely the two daughter cells will contain the plasmid. Generally, each molecule of plasmid diffuses randomly, so the probability of having a plasmid-less daughter cell is 21−N, where N is the number of copies. For instance, if there are 2 copies of a plasmid in a cell, there is a 50% chance of having one plasmid-less daughter cell. However, high-copy number plasmids have a cost for the hosting cell. This metabolic burden is lower for low-copy plasmids, but those have a higher probability of plasmid loss after a few generations. To control vertical transmission of plasmids, in addition to controlled-replication systems, bacterial plasmids use different maintenance strategies, such as multimer resolution systems, ...
Plasmid-mediated resistance is the transfer of antibiotic resistance genes which are carried on plasmids. The plasmids can be transferred between bacteria within the same species or between different species via conjugation. Plasmids often carry multiple antibiotic resistance genes, contributing to the spread of multidrug-resistance (MDR). Antibiotic resistance mediated by MDR plasmids severely limits the treatment options for the infections caused by Gram-negative bacteria, especially Enterobacteriaceae family. The global spread of MDR plasmids has been enhanced by selective pressure from antibiotic usage in human and veterinary medicine. Resistance plasmids by definition carry one or more antibiotic resistance genes. They are frequently accompanied by the genes encoding virulence determinants, specific enzymes or resistance to toxic heavy metals. Multiple resistance genes are commonly arranged in the resistance cassettes. The antibiotic resistance genes found on the plasmids confer resistance ...
As I understand it, plasmids, like mitochondria, have their own genetic material and are capable of self-replication. According to Wikipedia: Plasmids are considered replicons, units of DNA capable of replicating autonomously within a suitable host. However, like viruses, they are not classified as life. Plasmids are transmitted from one bacterium to another through conjugation. Unlike viruses, plasmids are naked DNA. However, some classes of plasmids encode the conjugative sex pilus necessary for their own transfer. My understanding of that is that a bacteria gets their plasmids not because of the replication of their circular chromosome, nor because that chromosome have genes to code for the plasmid (I dont really know if thats possible), but because of the self-replication of their own plasmids. So, my question is how the first plasmid got into the first bacteria, if they are not in their chromosomes? Were they a virus other prokaryotic cell that had circular DNA, and got phagocytosed ...
Fragments ofCandida boidinii chromosomal DNA were inserted into the integrative vector YIp-kanr and examined for the presence of sequences promoting autonomous replication of plasmids inSaccharomyces cerevisiae. Restriction maps of two plasmids, designated S6/4 and S6/5, originating from the sameS. cerevisiae transformant, were constructed. Southern hybridization data confirmed that the plasmids carry sequences from theC. boidinii chromosome. Both plasmids transformS. cerevisiae strains at 4-5-fold higher frequency than cloning vectors based on the replication origin of the 2μm plasmid. Mitotic stability of the constructed plasmids is similar to that of the 2μ-based vector pNF2 inS. cerevisiae.
To study thermal adaptations in the cyanobacterium, Synechococcus sp. PCC 7002, we screened about 3,000 mutants for their tolerance to high temperature, and found one, SHT1, that is sensitive to high-temperature stress. The mutant had a modified gene construct in the endogenous plasmid, pAQ1. One of the four ORFs, ORF93, was duplicated, and its mRNA level was higher than in the wild type. At 38°C, the growth of SHT1 was retarded as compared with the wild type, and above 38°C, almost all the cells of SHT1 died. This temperature is much lower than that required for induction of heat shock proteins. Interestingly, in both the wild type and SHT1, the thermal stability of oxygen-evolving machinery increased upon acclimation to high temperatures. These findings indicate that the lack of thermal tolerance in the SHT1 strain is likely independent of the adaptation of the PSII complex and heat shock responses, whereas there are essential contributions of genes in the endogenous plasmid to the ...
SUMMARY: Phylogenetic and epidemiological relatedness among transferable plasmids belonging to the IncC, IncM and IncH incompatibility groups has been studied by DNA-DNA filter hybridization. Hybridization was carried out on nitrocellulose microfilters, at low temperature, in formamide and under paraffin oil. The degree of hybridization among plasmids belonging to the IncC and IncM groups supported the conclusions drawn from genetic classification. Studies on relatedness among plasmids belonging to the IncH group allowed their classification into three phylogenetic sub-groups. Comparison of DNA sequences of three plasmids sharing the same genetic properties and isolated from different bacterial species suggested an epidemiological spread of the same plasmid.
Recreate original plasmid by cutting out insert - posted in Molecular Cloning: Maybe this is a stupid question, but I am going to ask it anyway: I would like to cut out an insert from my plasmid (pET28c) between restriction sites Nhe1 and Not1 and ligate the plasmid back to its orginial state without the insert (no I dont have the original plasmid anymore). However, obviously these restriction sites dont generate compatible sticky ends. Does anyone know how to recreate this...
Objectives: To investigate the genetic features of three plasmids recovered from an MCR-1 and ESBL-producing Escherichia coli strain, HYEC7, and characterize the transmission mechanism of mcr-1. Methods: The genetic profiles of three plasmids were determined by PCR, S1-PFGE, Southern hybridization and WGS analysis. The ability of the mcr-1-bearing plasmid to undergo conjugation was also assessed. The mcr-1-bearing transposon Tn6330 was characterized by PCR and DNA sequencing. Results: Complete sequences of three plasmids were obtained. A non-conjugative phage P7-like plasmid, pHYEC7-mcr1, was found to harbour the mcr-1-bearing transposon Tn6330, which could be excised from the plasmid by generating a circular intermediate harbouring mcr-1 and the ISApl1 element. The insertion of the circular intermediate into another plasmid, pHYEC7-IncHI2, could form pHNSHP45-2, the original IncHI2-type mcr-1-carrying plasmid that was reported. The third plasmid, pHYEC7-110, harboured two replicons, IncX1 and ...
Despite the near-ubiquity of plasmids in bacterial populations and the profound contribution of plasmid-borne genes and infectious gene transfer to the adaptation and evolution of bacteria, the mechanisms responsible for the maintenance of plasmids in bacterial populations are poorly understood. In this report, we address the question of how plasmids manage to persist over evolutionary time. Previous explanations have typically relied upon the ability of plasmids to deliver occasionally-useful genes to the right place at the right time. In contrast, we present a general mathematical proof that if (as suggested by several empirical studies) plasmids are not infectiously transmitted at a rate high enough to be maintained as genetic parasites in single populations, they will not be able to persist indefinately in these populations by carrying genes that are beneficial or sometimes beneficial to their host bacteria. Using more specific mathematical models, along with computer simulations, we ...
uncut plasmid dna vs linearized plasmid gel - posted in Molecular Cloning: Hello, I am going to run a gel comparing my uncut plasmid dna vs linearized plasmid. My insert is 135 bps and my vector is 3Kb. What can I expect to see on my gel, and how many bands can I expect to see. I am assuming the uncut plasmid will have several bands at different sizes and the linearized will have only one, is that right ? I am assuming the total size of my product will now be 3135 bps. Pls advise.
Figure 20. The pBR322 plasmid. Two genes of pBR322 confer resistance to antibiotics to any cell that contains the plasmid. AmpR confers resistance to amplicillin and tetR confers resistance to tetracycline to cells containing the plasmids. AmpR is a gene that codes for the periplasmic enzyme beta-lactamase that cleaves the ring structure found in amphicillin, which is a penicillin antibiotic. TetR is a gene that codes for a protein that modifies the bacterial cell wall and prevents tetracycline from entering the cell.. Multiple restriction endonuclease sites are present where foreign DNA fragments may be inserted.. Relaxed plasmid DNA replication continues in the presence of chloramphenicol. An interesting feature of this plasmid is that relaxed plasmid DNA replication continues even in the presence of an inhibitor of protein synthesis such as chloramphenicol. This feature allows increased yields of plasmid/cell of up to 100-fold ...
Easy Yeast Plasmid Isolation Kit: rescue plasmids from yeast using spin columns. Simple and highly efficient method. Zymolyase included.
One of the disadvantages of circular plasmids and chromosomes is their high sensitivity to rearrangements caused by homologous recombination. Odd numbers of crossing-over occurring during or after replication of a circular replicon result in the formation of a dimeric molecule in which the two copies of the replicon are fused. If they are not converted back to monomers, the dimers of replicons may fail to correctly segregate at the time of cell division. Resolution of multimeric forms of circular plasmids and chromosomes is mediated by site-specific recombination, and the enzymes that catalyze this type of reaction fall into two families of proteins: the serine and tyrosine recombinase families. Here we give an overview of the variety of site-specific resolution systems found on circular plasmids and chromosomes.
Santa Cruz Biotechnology now offers target-specific CRISPR/Cas9 Knockout (KO) Plasmids, CRISPR Double Nickase Plasmids and CRISPR/dCas9 Activation Plasmids for over 18,910 human and 18,340 mouse protein encoding genes. CRISPR/Cas9 KO Plasmids and CRISPR Double Nickase Plasmids enable the identification and cleavage of specific genes encoding a protein of interest thereby eliminating production of that gene product (protein). CRISPR/dCas9 Activation Plasmids activate endogenous gene transcription using a robust synergistic activation mediator (SAM) system. This exciting new technology is a useful tool for studying protein function and signalling pathways ...
Studying the transfer of specific mobile genetic elements in complex environmental matrices remains difficult because suitable molecular tools are not yet available to back up classical culture-dependent approaches. In this report, we show that quantitative PCR could be used to monitor the dissemination of the broad-host-range plasmid pB10 in sediment microcosms. This approach lies in the differential measurement of the host and plasmid DNAs used to inoculate the microcosms, using a particular design of quantitative PCR primers/probes where we took advantage of the mosaic aspect of the bacterial genomes to achieve a highly specific quantitative PCR detection system.
The segregational stability of bacterial, low-copy-number plasmids is promoted primarily by active partition. The plasmid-specified components of the prototypical P1 plasmid partition system consist of two proteins, ParA (44.3 kDa) and ParB (38.5 kDa), which, in conjunction with integration host fac …
In spite of the importance of plasmids in bacterial adaptation we have a poor understanding of their dynamics. It is not known if or how plasmids persist in and spread through (invade) a bacterial population when there is no selection for plasmid-encoded traits. Moreover, the differences in dynamics between spatially structured and mixed populations are poorly understood. Through a joint experimental/theoretical approach we tested the hypothesis that self-transmissible IncP-1 plasmids can invade a bacterial population in the absence of selection when initially very rare, but only in spatially structured habitats and when nutrients are regularly replenished. Using protocols that differed in degree of spatial structure and nutrient levels, the invasiveness of plasmid pB10 in E. coli was monitored during at least 15 days, with an initial fraction of plasmid-bearing (p+) cells as low as 1E-7. To further explore the mechanisms underlying plasmid dynamics we developed a spatially explicit mathematical ...
Mono- and Stereopictres of 5.0 Angstrom coordination sphere of Manganese atom in PDB 3dkx: Crystal Structure of the Replication Initiator Protein Encoded on Plasmid PMV158 (Repb), Trigonal Form, to 2.7 Ang Resolution
Difficult to Express Proteins: Novel Plasmid Technology to Significantly Increase Product Yield in CHO Cells, by Marco Cacciutolo
Mcr-1-harboring Enterobacteriaceae are reported worldwide since their first discovery in 2015. However, a limited number of studies are available that compared full-length plasmid sequences of human and animal origins. In this study, mcr-1-bearing plasmids from seven Escherichia coli isolates recovered from patients (n = 3), poultry meat (n = 2) and turkey meat (n = 2) in Switzerland were further analyzed and compared. Isolates were characterized by multilocus sequence typing (MLST). The mcr-1-bearing plasmids were transferred by transformation into reference strain E. coli DH5α and MCR-1-producing transformants were selected on LB-agar supplemented with 2 mg/L colistin. Purified plasmids were then sequenced and compared. MLST revealed six distinct STs, illustrating the high clonal diversity among mcr-1-positive E. coli isolates of different origins. Two different mcr-1-positive plasmids were identified from a single E. coli ST48 human isolate. All other isolates possessed a
The reason that we stock plasmids that only contain one terminator is because sometimes, despite the increased versatility, researchers may not want unnecessary terminators in their plasmids, for example when size constraints are an issue. The button below takes you to the plasmids with single terminators. If you prefer to use a plasmid with triple terminator, we suggest you search using the Plasmid Seach tool since nearly all our plasmids have triple terminators. The Plasmid Search tool will give you access to our full plasmid catalogue.. Finally, please try the Design Your Own Plasmid Online button below, to see what our cloning system can do for you. And remember that, while our plasmids are designed for easy cloning modifications, we are happy to do it for you if you prefer to outsource the cloning work.. ...
Prokaryotic transcriptomes change not only in response to physiological parameters but also to genetic rearrangements mediated by mobile elements. Plasmids are extrachromosomal genetic elements that replicate autonomously, and many can be transmitted between different strains through conjugation. Plasmids provide benefits to their hosts, such as resistance to antibiotics or degradation of recalcitrant aromatic compounds [1]; however, in several cases, the carriage of a large plasmid results in changes in the transcriptome of the host chromosome [2-4]. Similar to the effects of plasmid carriage on the transcriptional network of the host chromosome, differences in host background can alter the transcription patterns of backbone and accessory genes on a plasmid. Many plasmid backbone genes essential for conjugative transfer, replication initiation, and active partitioning are regulated both autogenously and by host factors [5]. Additionally, a number of plasmid-encoded degradative accessory genes ...
Synthetic biology heavily depends on rapid and simple techniques for DNA engineering, such as Ligase Cycling Reaction (LCR), Gibson assembly and Golden Gate assembly, all of which allow for fast, multi-fragment DNA assembly. A major enhancement of Golden Gate assembly is represented by the Modular Cloning (MoClo) system that allows for simple library propagation and combinatorial construction of genetic circuits from reusable parts. Yet, one limitation of the MoClo system is that all circuits are assembled in low- and medium copy plasmids, while a rapid route to chromosomal integration is lacking. To overcome this bottleneck, here we took advantage of the conditional-replication, integration, and modular (CRIM) plasmids, which can be integrated in single copies into the chromosome of Escherichia coli and related bacteria by site-specific recombination at different phage attachment (att) sites. By combining the modularity of the MoClo system with the CRIM plasmids features we created a set of 32 novel
ID YEP367 preliminary; circular DNA; SYN; 8400 BP. XX AC ATCC37735; XX DT 01-JUL-1993 (Rel. 7, Created) DT 01-JUL-1995 (Rel. 12, Last updated, Version 1) XX DE Saccharomyces/E.coli plasmid vector YEp367 - incomplete. XX KW cloning vector. XX OS Cloning vector OC Artificial sequences; Cloning vehicles. XX RN [1] RC YEp352E from YEp352 & linker RC YEp363A from pNM480 & YEp351 RC YEp353A from pNM480 & YEp352 RC YEp353 from YEp353A & YEp352E RC YEp354A from pNM481 & YEp352 RC YEp354 from YEp354A & YEp352E RC YEp355A from pNM482 & YEp352 RC YEp355 from YEp355A & YEp352E RC YEp356, YEp356R from YEp353 & pUC18 RC YEp357, YEp357R from YEp354 & pUC18 RC YEp358, YEp358R from YEp355 & pUC18 RC YEp363 from YEp363A & YEp353 RC YEp364 from YEp363A & YEp354 RC YEp365 from YEp363A & YEp355 RC YEp366 from YEp363A & YEp356 RC YEp367 from YEp363A & YEp357 RC YEp368 from YEp363A & YEp358 RC YEp366R from YEp363A & YEp356R RC YEp367R from YEp363A & YEp357R RC YEp368R from YEp363A & YEp358R RC YIp353 from YEp353 & ...
Background Prokaryotic plasmids have played out significant roles in the evolution of bacterial genomes and have a great impact on the metabolic functions of the host cell. variable genes (distributed genes and unique genes) than to the chromosomal core genes. Although all the functional categories of the chromosomal genes were exhibited by the plasmid genes, the proportions of each category differed between these two gene sets. The 598 gene families shared between chromosomes and plasmids displayed a uniform distribution between the two groups. A phylogenetic analysis of the shared genes, including the chromosomal core gene set, indicated that gene exchange events between plasmids and chromosomes occurred frequently during the evolutionary histories of the strains and species in this group. Moreover, the shared genes between plasmids and chromosomes usually had different promoter and terminator sequences, suggesting that they are regulated by different elements at the transcriptional level. ...
www.MOLUNA.de Plasmids in Bacteria [4191995] - Structure and Evolution.- Plasmids as Organisms.- Report on a Workshop: Structure and Function.- Evolutionary Relevance of Genetic Rearrangements Involving Plasmids.- Mechanisms of Transposition in Bacteria.- Insertion of Transcriptional Elements Outside the Replication Region Can Interfere with Replication, Maintenance, and Stability of ColE1-Derived Plasmids.- Studies on the Transposition of IS1.- On
ID YEP353 preliminary; circular DNA; SYN; 7944 BP. XX AC U03500; ATCC37725; XX DT 01-JUL-1993 (Rel. 7, Created) DT 01-JUL-1995 (Rel. 12, Last updated, Version 1) XX DE Saccharomyces/E.coli plasmid vector YEp353 - complete. XX KW cloning vector. XX OS Cloning vector OC Artificial sequences; Cloning vehicles. XX RN [1] RP 1-7944 RC YEp352E from YEp352 & linker RC YEp363A from pNM480 & YEp351 RC YEp353A from pNM480 & YEp352 RC YEp353 from YEp353A & YEp352E RC YEp354A from pNM481 & YEp352 RC YEp354 from YEp354A & YEp352E RC YEp355A from pNM482 & YEp352 RC YEp355 from YEp355A & YEp352E RC YEp356, YEp356R from YEp353 & pUC18 RC YEp357, YEp357R from YEp354 & pUC18 RC YEp358, YEp358R from YEp355 & pUC18 RC YEp363 from YEp363A & YEp353 RC YEp364 from YEp363A & YEp354 RC YEp365 from YEp363A & YEp355 RC YEp366 from YEp363A & YEp356 RC YEp367 from YEp363A & YEp357 RC YEp368 from YEp363A & YEp358 RC YEp366R from YEp363A & YEp356R RC YEp367R from YEp363A & YEp357R RC YEp368R from YEp363A & YEp358R RC YIp353 ...
DNA plasmids. Coloured atomic force micrograph of numerous pGL3 plasmids of DNA (deoxyribonucleic acid). A plasmid is a loop of DNA that can exist in a cell independently of the cells chromosomal DNA. Plasmids are important as they can replicate and express genes despite being outside the organisms chromosomes. This means that they can be used to introduce genes to organisms, a process used in gene therapy and genetic engineering. Genes, specific lengths of DNA, determine the development and characteristics of an organism. Atomic force microscopes make an image by moving a sensitive probe over a surface. Magnification: x24,000 at 6x7cm size. - Stock Image G110/0752
The present results concern the recombinant bacteria Escherichia coliHB101(GAPDH) which produces glyceraldehyde 3-phosphate dehydrogenase. An unusual phenomenon was noticed concerning the plasmid...
Many bacteria carry extra DNA molecules beyond their chromosome, so-called plasmids. While plasmids are apriori costly to the cell, they ...
The combination of a p15A and a colE1 origin of replication plasmid is the most common two-plasmid system for use in E. coli. The pSB1A* series of plasmids contain the colE1 plasmid and cannot be co-maintained at stable copy numbers in combination due to plasmid incompatibility. Here, two plasmids in the same cell with origins from the same incompatibility group compete for replication machinery, resulting in unpredictable copy number and often the exclusion of one of the plasmids. This is circumvented by cotransforming plasmids with origins from distinct incompatibility groups. Since no p15A origin plasmid was present in the Registry, we developed part J23001, or pSB3C6 for our studies. This plasmid was used for all our riboregulator experiments for the locked-RFP reporter. Shown here is the variant of the plasmid containing part J01022. Plasmids for constructing basic Biobrick parts ...
TY - JOUR. T1 - Comparative analysis of conjugative plasmids mediating gentamicin resistance in Staphylococcus aureus. AU - Goering, R. V.. AU - Ruff, E. A.. PY - 1983/1/1. Y1 - 1983/1/1. N2 - Five gentamicin-resistant clinical isolates of Staphylococcus aureus were found to contain self-transmissible plasmids of 32 to 37 megadaltons in size. Restriction endonuclease digests of the plasmids were markedly similar to those of reference plasmids of unrelated geographical origin, thus suggesting the significant contribution of common conjugal plasmids to the emergence of gentamicin resistance in S. aureus populations.. AB - Five gentamicin-resistant clinical isolates of Staphylococcus aureus were found to contain self-transmissible plasmids of 32 to 37 megadaltons in size. Restriction endonuclease digests of the plasmids were markedly similar to those of reference plasmids of unrelated geographical origin, thus suggesting the significant contribution of common conjugal plasmids to the emergence of ...
Objectives: To investigate the genetic features of five plasmids recovered from an NDM-5-producing clinical Escherichia coli strain, BJ114, and to characterize the plasmid recombination event that occurred during the conjugation process. Methods: The genetic profiles of the five plasmids were determined by PCR, conjugation, S1-PFGE, Southern hybridization and WGS analysis. Plasmid sequences were analysed with various bioinformatic tools. Results: Complete sequences of five plasmids were obtained. Two small plasmids, pBJ114-141 and pBJ114-46, were speculated to have recombined into a large fusion plasmid, pBJ114T-190. When conjugated to other E. coli strains, some of the fusion plasmids were able to be resolved into the original two single plasmids. A nonconjugative plasmid, pBJ114-96, exhibited a high degree of sequence identity with the phage P7-like plasmid as well as an mcr-1-bearing plasmid. Another plasmid, pBJ114-78, was found to contain multidrug resistance genes and various mobile ...
© 2018 Macmillan Publishers Ltd., part of Springer Nature. Plasmids have a major role in the development of disease caused by enteric bacterial pathogens. Virulence plasmids are usually large (|40 kb) low copy elements and encode genes that promote host-pathogen interactions. Although virulence plasmids provide advantages to bacteria in specific conditions, they often impose fitness costs on their host. In this Review, we discuss virulence plasmids in Enterobacteriaceae that are important causes of diarrhoea in humans, Shigella spp., Salmonella spp., Yersinia spp and pathovars of Escherichia coli. We contrast these plasmids with those that are routinely used in the laboratory and outline the mechanisms by which virulence plasmids are maintained in bacterial populations. We highlight examples of virulence plasmids that encode multiple mechanisms for their maintenance (for example, toxin-antitoxin and partitioning systems) and speculate on how these might contribute to their propagation and success.
Biotechnical production processes often operate with plasmid-based expression systems in well-established prokaryotic and eukaryotic hosts such as Escherichia coli or Saccharomyces cerevisiae, respectively. Genetically engineered organisms produce important chemicals, biopolymers, biofuels and high-value proteins like insulin. In those bioprocesses plasmids in recombinant hosts have an essential impact on productivity. Plasmid-free cells lead to losses in the entire product recovery and decrease the profitability of the whole process. Use of antibiotics in industrial fermentations is not an applicable option to maintain plasmid stability. Especially in pharmaceutical or GMP-based fermentation processes, deployed antibiotics must be inactivated and removed. Several plasmid addiction systems (PAS) were described in the literature. However, not every system has reached a full applicable state. This review compares most known addiction systems and is focusing on biotechnical applications.. ...
TY - JOUR. T1 - Restriction endonuclease analysis of Staphylococcus aureus plasmid DNA from three continents. AU - Doebbeling, Bradley. AU - Pfaller, M. A.. AU - Hollis, R. J.. AU - Boyken, L. D.. AU - Pignatari, A. C.. AU - Herwaldt, L. A.. AU - Wenzel, R. P.. PY - 1992/1. Y1 - 1992/1. N2 - Staphylococcus aureus isolates (n=1201) from 20 centers in Europe, the USA and Brazil were evaluated for the presence of epidemiologic markers. Plasmid typing and restriction endonuclease analysis of plasmid DNA confirmed the presence of an apparently identical plasmid in 13 % of clinical isolates. The plasmid was recovered from all 20 hospitals studied, with an overall frequency of ,10 % on each of the three continents. Since relatively few staphylococcal plasmids may be shared by epidemiologically unrelated strains, there are inherent limitations to this otherwise useful technique. Additionally, these data demonstrate the importance of including unrelated strains of Staphylococcus aureus from the local ...
To further characterize the fosB-carrying plasmids of 19 vancomycin-resistant enterococci, the complete sequences of the fosB- and vanA-containing plasmids of E. faecium (pEMA120) and E. avium (pEA19081) were obtained by single-molecule, real-time sequencing. We found that these two plasmids are essentially identical (99.99% nucleotide sequence identity), which proved the possibility of interspecies transmission. Comparative analysis of the plasmids revealed that the backbone of pEMA120 is 99% similar to a conjugative fosB-negative E. faecium plasmid, pZB18. There is a traE disrupted in the transfer region of pEMA120, in comparison to pZB18 with an intact traE. The difference of their transfer frequencies between pEMA120 and pZB18 suggests this interruption of traE might affect conjugative transfer. Two copies of the fosB gene linked to a tnpA gene, forming an ISL3-like transposon, were found at separate locations within pEMA120, which had not been reported previously. These two fosB-carrying
The sequence analysis of the 7383 bp plasmid pCIZ2 from |i|Enterococcus faecium|/i| L50 enabled the identification of a DNA region involved in its replication. The structural organization of the pCIZ2 replication region is highly similar to those of well-known theta-replicating plasmids. It contains an untranslated region, the putative replication origin (|i|ori|/i|), constituted by two sets of direct repeats of 12 and 22 bp (iterons), and followed by three open-reading frames (|i|orf8|/i| to |i|orf10|/i|). |i|orf8|/i| encodes the replication initiation protein (RepE). The transcriptional start site of the replication locus was identified 13 nucleotides upstream of the |i|repE|/i| start codon. A two-dimensional agarose gel electrophoresis analysis revealed pCIZ2 intermediates profile typical of the theta-type replication mechanism. Subcloning of different DNA fragments of the pCIZ2 replication region in |i|Escherichia coli|/i| and, subsequently, in the plasmidless |i|E. faecium|/i| L50/14-2 allowed the
Gene transfer mediated by plasmids plays an important role in the spread of antibiotic resistance genes. Nowadays, high molecular weight plasmids carrying resistance genes to antibiotics and growth inhibitors are commonly found in pathogenic bacteria. Moreover, conjugative plasmids coding for antibiotic resistance may interact with virulence plasmids thus resulting in plasmid transferring both antibiotic resistance and virulence genes. In this thesis I therefore pursued genomics and proteomics studies on plasmids of two incompatibility group. In the first part of the thesis we determined the complete nucleotide sequences of multidrug resistance plasmid p9134 and its two variants, p9134dT which spontaneously lost tetracycline resistance gene, and p9134dAT which spontaneously lost ampicillin and tetracycline resistance genes. Plasmids were 133 802 bp, 127 291 bp a 109 512 bp in size, respectively, and their basic backbone was similar to that of IncI plasmids. In plasmid p9134 genes coding for ...
ii) Evolution of the blaNDM-1 locus.The mer operon of pAR060302 is part of a complex hybrid transposon inserted into the IncA/C backbone flanked by direct repeats, and related structures are found in other IncA/C plasmids. The strA-strB genes in pYR1 are part of a Tn5393 that was recently described (3, 4). The synteny among the compared plasmids restarted at the DNA primase gene (Fig. 1). IncA/C plasmid pP91278 from P. damselae likely represents the IncA/C scaffold before the acquisition of the accessory genes, since it does not show any insertion between the phage-integrase, rhs, and DNA primase genes (Fig. 1).. An analysis of the pNDM-KN sequence revealed that the rhs gene was deleted at its 3′ and 5′ extremities and was adjacent to the heat shock chaperone groEL-groES cluster (28). In pAR060302, this cluster flanked a class 1 integron that contained the aac(3)-IVa and aadA1 gene cassettes, but it was absent in plasmids pYR1 and pP91278 (Fig. 1). Analysis of the sequences located at the ...
Integrative and conjugative elements (ICEs, also known as conjugative transposons) are mobile elements that are found integrated in a host genome and can excise and transfer to recipient cells via conjugation. ICEs and conjugative plasmids are found in many bacteria and are important agents of horizontal gene transfer and microbial evolution. Conjugative elements are capable of self-transfer and also capable of mobilizing other DNA elements that are not able to self-transfer. Plasmids that can be mobilized by conjugative elements are generally thought to contain an origin of transfer (oriT), from which mobilization initiates, and to encode a mobilization protein (Mob, a relaxase) that nicks a site in oriT and covalently attaches to the DNA to be transferred. Plasmids that do not have both an oriT and a cognate mob are thought to be nonmobilizable. We found that Bacillus subtilis carrying the integrative and conjugative element ICEBs1 can transfer three different plasmids to recipient bacteria at ...
When you want sustained transgene expression without introducing any foreign DNA-such as for model animal and gene therapy development-Minicircle Technology is a great gene expression option. Produced as small excised, circular DNA fragments from a parental plasmid, the non-viral, episomal Minicircle expression cassette is free of any bacterial plasmid DNA sequences, and comes with a variety of promoter and reporter combinations. Their small size facilitates more efficient transfection than whats possible with standard-sized plasmids, and, while Minicircles do not replicate with the host cell, expression lasts for 14 days or longer in dividing cells, and can continue for months in non-dividing cells.. Product Note:. Parental minicircle plasmids and the ZYCY10P3S2T Producer Bacterial Strain are available for purchase by not-for-profit researchers only. Commercial users may purchase pre-made, ready-to-transfect minicircle DNA only. SBI also offers custom parental plasmid cloning and minicircle ...
|jats:title|ABSTRACT|/jats:title||jats:p|OXA-48-like enzymes have emerged as important extended-spectrum β-lactamases/carbapenemases in|jats:named-content xmlns:xlink=http://www.w3.org/1999/xlink content-type=genus-species xlink:type=simple|Escherichia coli|/jats:named-content|sequence type 131 (ST131). We report the structures of the first fully sequenced|jats:italic|bla|/jats:italic||jats:sub|OXA-163|/jats:sub|plasmid and of two other|jats:italic|bla|/jats:italic||jats:sub|OXA-48|/jats:sub|plasmids in this lineage.|jats:italic|bla|/jats:italic||jats:sub|OXA-163|/jats:sub|was located on a 71-kb IncN plasmid with other resistance genes.|jats:italic|bla|/jats:italic||jats:sub|OXA-48|/jats:sub|was present on IncL/M plasmids, genetically similar to other|jats:italic|bla|/jats:italic||jats:sub|OXA-48|/jats:sub|plasmid sequences, and consistent with interspecies/interlineage spread. The presence of|jats:italic|bla|/jats:italic||jats:sub|OXA-48-like|/jats:sub|genes on epidemic plasmids in ST131 is of
This chapter provides a broad overview of many applications of plasmids for genetic analysis, primarily in bacteria. Ever since DNA sequencing became accessible to most research laboratories, reverse genetic analysis has become a standard experimental approach to study bacterial gene function. Similar suicide vectors have also been used for nontargeted insertional mutagenesis by cloning random chromosomal DNA fragments into the plasmid. The use of suicide vectors also allows for easy identification of the insertion mutations. Plasmids that utilize different combinations of double-counter selective markers have been used for diverse applications, including the search for extremely rare suppressor mutations of essential Escherichia coli genes, and to improve the efficiency of allelic exchange on bacterial artificial chromosomes (BACs). Although temperature-sensitive vectors represent the majority of conditionally replicating plasmids, other plasmids that exhibit conditional replication have been described
Non-viral gene therapy is being considered as a treatment for cystic fibrosis. In clinical studies and in studies using the mouse airways as a model, current formulations result in only transient transgene expression. A number of reasons for this have been proposed including the loss of plasmid DNA from cells. The aim of these studies was to investigate why transgene expression from non-viral vectors is transient in the mouse lung. Plasmid DNA encoding the luciferase reporter gene was complexed with the cationic lipid GL67 and delivered to the mouse airways. The persistence of plasmid DNA in the mouse lungs was investigated using quantitative PCR and Southern hybridization. Results showed that intact plasmid DNA persisted in the mouse lung in the absence of any detectable luciferase activity. The de novo methylation of plasmid DNA in vivo was investigated as a potential cause of this transient gene expression but results suggested that plasmid DNA does not become de novo methylated in the mouse lung.
The conditions promoting the persistence of a plasmid carrying a trait that may be mutually beneficial to other cells in its vicinity were studied in structured and unstructured environments. A large plasmid encoding mercury resistance in Pseudomonas fluorescens was used, and the mercury concentration allowing invasion from rare for both plasmid-bearing and plasmid-free cells was determined for different initial inoculum densities in batch-culture structured (filter surface) and unstructured (mixed broth) environments. A range of mercury concentrations were found where both cell types could coexist, the regions being relatively similar in the two types of environment although density-dependent in the unstructured environment. The coexistence is explained in terms of frequency-dependent selection of the mutually beneficial mercury resistance trait, and the dynamics of bacterial growth under batch culture conditions. However, the region of coexistence was complicated by conjugation which increased plasmid
The International Society for Plasmid Biology and other Mobile Genetic Elements (ISPB) was established in 2004. The purpose of the ISPB is to promote the study of plasmids and other mobile genetic elements.. The Society will engage in collective activities that increase the awareness and understanding of the importance of these genetic elements both in the scientific community and in the public arena.. The Society will highlight important discoveries in the plasmid field as well as new and novel applications of plasmids and other mobile genetic elements, and will act as an advocate for increased research support of this field.. ISPB will also promote the importance of retaining the study of plasmids and other mobile genetic elements in undergraduate teaching curriculum.. The Society will provide a forum for the plasmid biology community to work with sequence databases to ensure the highest quality of annotation.. The constitution of ISPB can be found here.. Information about previous conferences ...
SUMMARY: A PUB 110-derived plasmid encoding chloramphenicol resistance, kanamycin resistance and high-temperature α-amylase showed a high degree of segregational instability when inserted into Bacillus subtilis. In an attempt to obtain stable derivatives, the organism was grown in chemostat culture in the presence of chloramphenicol. It was periodically found necessary to increase the concentration of chloramphenicol in the medium feed in order to avoid plasmid loss. Strains were isolated after 19 and 160 generations, which showed high levels of plasmid stability. This characteristic appeared to be genotypic. No detectable difference in plasmid copy number was found between the original and the improved strains. The stability characteristics resided in the host, rather than in the plasmid. Stable isolates possessed elevated MICs for both chloramphenicol and kanamycin. Their maximum specific growth rates were higher than that of the original strain, and similar to that of the plasmid-free parent strain.
BACKGROUND: Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery. METHODS: Enhanced electrically-mediated delivery, and less extensively, liposome complexed delivery, of a plasmid encoding the reporter luciferase was tested in rodent skin. Expression kinetics and tissue damage were explored as well as testing in a second rodent model. RESULTS: Experiments confirm that electroporation alone is more effective in enhancing reporter gene expression than plasmid injection alone, plasmid conjugation with liposomes followed by injection, or than the combination of liposomes and electroporation. However, with two time courses of multiple
Extended-spectrum cephalosporins have been classified by the World Health Organization as critically important antibiotics in human medicine (19). Ceftiofur is an extended-spectrum cephalosporin licensed for the treatment of respiratory infections in pigs and cattle. Various authors have hypothesized that the veterinary use of extended-spectrum cephalosporins may select for extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in animals, resulting in an increased risk of the zoonotic transmission of ESBL-carrying bacteria and plasmids (2, 10, 11, 17, 18).. A statistical association between the prophylactic use of ceftiofur and the occurrence of cefotaxime-resistant E. coli in healthy pigs was recently demonstrated at two Danish pig farms, and most of the isolates were shown to be CTX-M-1 producers (11). In this study, we revisited the farms 1 year after the previous study to investigate the distribution, persistence, and transmission of blaCTX-M-1 between pigs, farm workers, and ...
산타크루즈바이오테크놀러지는 광범위한 유전자편집 제품을 제공하고 있으며 유전자침묵에 쓰이는 CRISPR/Cas9 Knockout 와 CRISPR Double Nickase plasmids를 제공합니다. Ribosomal Protein L39L유전자침묵에는 Ribosomal Protein L39L CRISPR/Cas9 Knockout plasmids 와 Ribosomal Protein L39L Double Nickase Plasmids를 제공합니다. 또한 유전자활성화에 쓰이는 Ribosomal Protein L39L CRISPR/dCas9 Activation Plasmids와 CRISPR Lenti Activation Systems도 제공합니다. 유전자침묵과 활성화는 유전자연구에 유용하게 쓰이며 이는 항체와 결합하여 단백질의 검출에 유용하게 쓰입니다.
산타크루즈바이오테크놀러지는 광범위한 유전자편집 제품을 제공하고 있으며 유전자침묵에 쓰이는 CRISPR/Cas9 Knockout 와 CRISPR Double Nickase plasmids를 제공합니다. Ribosomal Protein L39유전자침묵에는 Ribosomal Protein L39 CRISPR/Cas9 Knockout plasmids 와 Ribosomal Protein L39 Double Nickase Plasmids를 제공합니다. 또한 유전자활성화에 쓰이는 Ribosomal Protein L39 CRISPR/dCas9 Activation Plasmids와 CRISPR Lenti Activation Systems도 제공합니다. 유전자침묵과 활성화는 유전자연구에 유용하게 쓰이며 이는 항체와 결합하여 단백질의 검출에 유용하게 쓰입니다.
Vol 9: Complete Nucleotide Sequence and Analysis of Two Conjugative Broad Host Range Plasmids from a Marine Microbial Biofilm.. This article is from PLoS ONE, volume 9.AbstractThe complete nucleotide sequence of plasmids pMCBF1 and pMCBF6 was deter. Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
TM 10 6670-277 13&P APPENDIX B MAINTENANCE ALLOCATION CHART Section I. INTRODUCTION B 1. General. a. This section provides a general explanation of all maintenance and repair functions authorized at various maintenance categories. b. The Maintenance Allocation Chart (MAC) in Section II designates overall authority and responsibility for the performance of maintenance functions on the identified end item or component. The application of the maintenance functions to the end item or component will be consistent with the capacities and capabilities of the designated maintenance categories. c. Section Ill lists the tools and test equipment (both special tools and common tool sets) required for each maintenance function as referenced from Section Il. d. Section IV contains supplemental instructions and explanatory notes for a particular maintenance function. B 2. Maintenance Functions. Maintenance functions will be limited to and defined as follows: a. Inspect. To determine the serviceability of an ...
We have previously shown that the intramuscular injection of naked plasmid DNA enables foreign gene expression in muscle. Further studies showed that the intravascular delivery of naked plasmid DNA enables high levels of expression not only in muscle but also in hepatocytes. For the liver, this tech …
Bacterial circular chromosomes replicate bidirectionally by the theta mechanism. Circular plasmids replicate by three general mechanisms: theta type, strand displacement, and rolling circle (43). Different machineries are involved in different plasmid replication mechanisms (43). For example, initiation of theta-type plasmid replication generally requires a plasmid-encoded initiator protein. In contrast, initiation of strand displacement replication requires the plasmid-encoded helicase, primase, and initiator protein, and initiation of rolling-circle replication requires the plasmid-encoded protein with DNA strand transferase enzymatic activity. The absence of the above essential proteins for the latter two mechanisms indicated that Pseudoalteromonas chromids replicate by the theta mechanism. Consistent with this, no such proteins in chromids from other bacteria have been reported. Considering the different replication factors involved in different mechanisms, a parsimony assumption is that ...
Objective: Carbapenem-producing organisms have spread worldwide and cause significant morbidity. Horizontal gene transfer of carbapenemases may play a role in this spread. Given that conjugation is influenced by a number of factors, we sought to perform a systematic analysis of blaKPC encoding plasmid transfer into multiple species using hospital isolates. Methods: Plasmids were isolated from patient donor strains from the NIH and University of Virginia and a subset were tagged with GFP and electroporated into a K. pneumoniae patient isolate cured of its blaKPC plasmid. Broth and filter matings were performed, and transconjugants were isolated on selective media. Plasmids tested included those found in multiple species during hospital surveillance. Results: Transfer frequency was dependent on the recipient, temperature, substratum, and the specific plasmid. pKPC-47e was extremely attenuated in conjugation efficiency across all conditions tested compared to pKpQIL. In vitro studies showed a low ...
Interpretive Summary: A novel plasmid (pHoss1) for Listeria monocytogenes was constructed. pHoss1 is very efficient for the generation of in-frame deletion mutants. The L. monocytogenes ispG and ispH genes were in-frame mutated. The L. monocytogenes ispG and ispH genes are not involved in cell adhesion. Technical Abstract: Although several plasmids have been used in Listeria monocytogenes for generating mutants by allelic exchange, construction of L. monocytogenes mutants has been inefficient due to lack of effective selection markers for first and second recombination events. To address this problem, we have developed a new suicide plasmid, pHoss1, by using the pMAD plasmid backbone and anhydrotetracycline selection marker (secY antisense RNA) driven by an inducible Pxyl/tetO promoter. Expression of the secY antisense RNA eliminates merodiploids and selects for the loss of plasmid via a second allelic exchange, which enriches the number of mutants with deleted genes. To assess the effectiveness ...
In the PBE lab we study the role of plasmids as catalysts of bacterial evolution, with a special focus on the evolution of plasmid-mediated antibiotic resistance.. Currently, we have two ongoing projects:. - In vivo evolution of plasmid-mediated resistance.. Conjugative plasmids play a key role in the horizontal spread of antibiotic resistance mechanisms among bacteria. One of the key factors undermining the successful spread of a conjugative plasmid is the initial fitness cost produced by the plasmid in the recipient bacteria. The factors involved in this cost and its potential compensation remain largely unknown. In our lab we are trying to understand the evolutionary and genetic determinants that promote the emergence and establishment of successful associations between bacterial clones and resistance plasmids in vivo.. To do so we study conjugation events between different enterobacteria occurring in the gut of hospitalized patients. We study the cost produced by the plasmids when they first ...
Tagging proteins is a standard method facilitating protein detection, purification or targeting. When tagging a certain protein of interest, it is challenging to predict which tag will give optimal results and will not interfere with protein folding, activity or yields. Ideally, multiple tags and positions are tested which however complicates molecular cloning and expression vector generation. In conventional cloning, tags are either added on PCR primers (requiring a distinct primer and PCR product per tag) or provided on the vector (typically leaving a restriction site scar). Here we report a vector family of 40 plasmids allowing simple, seamless fusions of a single PCR product with various N- and C-terminal tags, signal sequences and promoters. The restriction site free cloning (RSFC) strategy presented in this paper relies on seamless cloning using type IIS restriction endonucleases. After cutting out a stuffer (placeholder) fragment from the vectors, a single PCR product can be directly inserted in
The F plasmid origin needs to be designed. The complete F plasmid with partitioning genes in ~10kb in length. It contains several BioBricks restriction sites in both coding and noncoding regions. Once designed, the F plasmid origin can be assembled with an antibiotic resistance marker and cloned into the vector scaffold to generate a new single copy BioBricks plasmid. Chris Anderson suggested inclusion of the R6K origin in these plasmids (rather than inclusion of a pUC19 origin in the multiple cloning site). The R6K origin is a conditional origin. It only works in the presence of the trans-acting protein Π (encoded by pir) for replication. R6K replicates at a medium copy (15 per cell) in pir+ strains and high copy (250 per cell) in pir-116 (high-copy-number mutant) E. coli hosts. ...
The F plasmid origin needs to be designed. The complete F plasmid with partitioning genes in ~10kb in length. It contains several BioBricks restriction sites in both coding and noncoding regions. Once designed, the F plasmid origin can be assembled with an antibiotic resistance marker and cloned into the vector scaffold to generate a new single copy BioBricks plasmid. Chris Anderson suggested inclusion of the R6K origin in these plasmids (rather than inclusion of a pUC19 origin in the multiple cloning site). The R6K origin is a conditional origin. It only works in the presence of the trans-acting protein Π (encoded by pir) for replication. R6K replicates at a medium copy (15 per cell) in pir+ strains and high copy (250 per cell) in pir-116 (high-copy-number mutant) E. coli hosts. ...
Results: Our goal was to develop a high-throughput recombineering system, primarily for the coupling of genes to epitope tags, which could also be used for deletion of genes in both pathogenic and K-12 E. coli strains. To that end we have designed a series of donor plasmids for use with the lambda-Red recombination system, which when cleaved in vivo by the I-SceI meganuclease generate a discrete linear DNA fragment, allowing for C-terminal tagging of chromosomal genes with a 6xHis, 3xFLAG, 4xProteinA or GFP tag or for the deletion of chromosomal regions. We have enhanced existing protocols and technologies by inclusion of a cassette conferring kanamycin resistance and, crucially, by including the sacB gene on the donor plasmid, so that all but true recombinants are counter-selected on kanamycin and sucrose containing media, thus eliminating the need for extensive screening. This method has the added advantage of limiting the exposure of cells to the potential damaging effects of the lambda-Red ...
This topic contains 2 study abstracts on Vaccination: Plasmid DNA Vaccines indicating they may negatively impact Infertility, Vaccination: Abortion, and Vaccine-induced Toxicity
The pCMV-GLuc 2 Control Plasmid is a mammalian expression vector that encodes the secreted luciferase from the copepod Gaussia princeps as a reporter, under the control of the constitutive CMV (cytomegalovirus) promoter. Gaussia luciferase (GLuc) is a 19 kDa protein encoded by a humanized sequence, and it contains a native signal peptide at the N-terminus that allows it to be secreted from mammalian cells into the cell culture medium. A neomycin resistance gene under the control of an SV40 promoter allows selection for stable integration of the plasmid into the mammalian cell genome using G418.
The mitochondrial gene oxi1, carried on a bacterial plasmid, has been used to transform the mitochondria of a yeast strain lacking mtDNA (rho0). The plasmid DNA behaved in a manner entirely consistent with the known properties of normal yeast rho- mtDNA after its introduction by high-velocity microprojectile bombardment. Like the mtDNA sequences retained in natural rho- strains, the plasmid DNA in the transformants was reiterated into concatemers whose size was indistinguishable from that of wild-type mtDNA. The oxi1 sequences in the transformants were surrounded by restriction sites derived from the plasmid that were not present in wild-type mtDNA. oxi1 genetic information in these synthetic rho- strains could be expressed in diploids either after marker rescue by recombination with rho+ mtDNA carrying an appropriate oxi1 point mutation or in trans during the growth of diploids heteroplasmic for both the plasmid-derived oxi1 sequences and rho+ mtDNA with oxi1 deleted. The ability to ...
I was hoping for suggestions as I am having difficulty getting two plasmids into E.coli at library efficiences. I am hoping to add a M13 derived vector to cells containing a pUC/pBR3222 plasmid, or vice versa. Electroporating the M13 derived vector has been at low efficiency so far, making double electroporations very inefficient. The cells containing these plasmids were less electrocompetent, less than 10^6 col/ ug of the second plasmid. Therefore, this approach would require many electroporations to get a large library. I next looked to make the M13 derived vector infectious by growing with helper phage, but had too much cross over with the helper phage. I am now trying this in a Rec minus strain. I was hoping to electroporate in one plasmid and then add the other plasmid through lambda infection but was unable to locate a non lysing system. Please let me know if you have had similar experiences or any suggestions. Thanks tj t j cradick ucsf dept of immunology please reply to:tjc at ...
The procedure has been used successfully for isolation of high- and low-copy-number plasmids from various Bacillus subtilis strains. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture ...
In the PBE lab we are interested in the evolutionary forces that drive plasmid dynamics in bacterial populations. Plasmids play a crucial role in bacterial ecology and evolution because they can transfer genes horizontally between different bacteria. The most striking example of how plasmids drive bacterial evolution is the global spread of plasmid-mediated antibiotic resistance over the last few decades. Plasmids are arguably the main vehicle for the spread of antibiotic resistance genes among clinically relevant bacteria, contributing to the overwhelming antibiotic resistance crisis we are currently facing.. In our group we try to understand the population genetics of antibiotic resistance plasmids using advanced molecular and evolutionary techniques. Ultimately, we intend to apply the concepts that we learn from the study of the evolution of plasmid-mediated antibiotic resistance to develop more rational intervention strategies to control infectious diseases.. ...
The same plasmid carrying blaCTX-M-14b was identified from an Escherichia coli isolate and an Enterobacter cloacae isolate collected from cattle in the United Kingdom by complete plasmid sequencing. This 35,341-bp plasmid, pSAM7, had an IncX4 backbone that is 99% identical to that of pJIE143 from a human isolate in Australia. PCR screening identified pSAM7-like plasmids in three other E. coli isolates of different multilocus sequence types isolated from cattle on different farms in the United Kingdom.. ...
pRV500 is, to our knowledge, the first plasmid of L. sakei to be entirely sequenced. Careful sequence analysis showed that pRV500 belongs to the pUCL287 family, itself related to class A theta-type plasmids of the pUCL22 and pSC101 families. Experimental evidence of the theta replication mode was obtained for pUCL287 itself (2). Although the minimum length of the pRV500 oriA region remains to be determined, it is likely to involve the same two kinds of DNA repeats as other pUCL287-type plasmids. The involvement of repetitive sequences of 22 bp (iterons) as a target for binding of the Rep protein was demonstrated for pSBO1 (40), and the upstream 11-bp repeated sequences were observed to be essential for pUCL287 replication (3).. Like other plasmids (14, 49), pRV500 appears to be a composite structure containing DNA segments from different sources. Some ORFs are shared by plasmids from both subfamilies pUCL22 and pUCL287; this applies to orf6 in pRV500 and ORFs of unknown function around the gene ...
Members of the family Enterobacteriaceae have been isolated from raw wastewater, identified, and characterized with respect to their plasmid content and antibiotic resistance. Several strains possessing both antibiotic resistance and high-molecular-weight plasmid(s) transferred their resistance characteristics to recipient cells during a 25 h coincubation. Eight were characterized (six Escherichia coli and two Klebsiella pneumoniae); each produced 10(2) to 10(7) transconjugants per ml by the end of the incubation period. They were also able to mobilize pBR325 from a laboratory E. coli strain into plasmid-free recipients to yield 10(2) to 10(7) transconjugants per ml. These transconjugants possessed phenotypic characteristics specified by pBR325, the R plasmid, and the chromosome of the recipient. Many transconjugants exhibited recombinational rearrangements of the acquired plasmid material. ...
Polymerase chain reaction-based replicon typing represents a novel method to describe the dissemination and follow the evolution of resistance plasmids. We used this approach to study 26 epidemiologically unrelated Enterobacteriaceae and demonstrate the dominance of incompatibility (Inc) A/C or Inc N-related plasmids carrying some emerging resistance determinants to extended-spectrum cephalosporins and carbapenems.
Bioz Stars score, Techniques, Protocol Conditions and more for Expression Plasmids, supplied by . Data for Expression Plasmids gathered from related PubMed articles.
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Plasmids can be broadly classified into conjugative plasmids and non-conjugative plasmids. Conjugative plasmids contain a set ... Many plasmids have been created over the years and researchers have given out plasmids to plasmid databases such as the non- ... Virulence plasmids, which turn the bacterium into a pathogen. e.g. Ti plasmid in Agrobacterium tumefaciens Plasmids can belong ... Plasmids are generally circular, but examples of linear plasmids are also known. These linear plasmids require specialized ...
The IncP-1 plasmid group (IncP plasmids in Escherichia coli) of which RK2 is a part has been described as "highly potent, self- ... The RK2 Plasmid is a broad-host-range plasmid belonging to the incP incompatibility group It is notable for its ability to ... Minimal plasmids such as PFF1 are useful for studying the basic mechanisms of plasmid replication and copy number regulation, ... Plasmid, 2010 Nov;64(3):119-34 U Kües and U Stahl: "Replication of plasmids in gram-negative bacteria", Microbiol Rev. 1989 ...
... is a non-coding RNA found in bacterial plasmids including pIP501. RNAIII acts by transcriptional attenuation of ... Page for Plasmid RNAIII at Rfam v t e (Non-coding RNA, All stub articles, Molecular and cellular biology stubs). ... importance of a U-turn loop structure in the target RNA of plasmid pIP501 for efficient inhibition by the antisense RNA". ...
... is a plasmid that contains an inserted piece of foreign DNA. "Everything Bio : Hybrid plasmid". Everythingbio. ... v t e (Articles needing additional references from November 2013, All articles needing additional references, Plasmids, All ...
A typical plasmid DNA yield of a miniprep is 5 to 50 µg depending on the cell strain. Miniprep of a large number of plasmids ... Minipreparation of plasmid DNA is a rapid, small-scale isolation of plasmid DNA from bacteria. It is based on the alkaline ... A plasmid preparation is a method of DNA extraction and purification for plasmid DNA. Many methods have been developed to ... The plasmid DNA yield will vary depending on the plasmid copy number, type and size, the bacterial strain, the growth ...
The R1 Plasmid is a plasmid that was first isolated from Salmonella paratyphi bacteria in 1963. It is a short plasmid, composed ... The R1 plasmid partitioning is a mechanism needed for the inheritance of the R1 plasmid. The par system is composed of the ParR ... The R1 plasmid imparts multi-drug antibiotic resistance to its host bacteria. It's known as a "low copy" plasmid, meaning that ... Cell division takes place, resulting in the partitioned plasmids in two daughter cells. Some genes on the R1 plasmid are: ParM ...
The Ti plasmid is a member of the RepABC plasmid family found in Alphaproteobacteria. These plasmids are often relatively large ... the Ti plasmid is part of a family of plasmids carried by many species of Alphaproteobacteria. Members of this plasmid family ... Some notable early milestones in the studies of Ti plasmids include the mapping of a Ti plasmid in 1978 and the studying of ... virF is a host specificity factor found in some but not all types of Ti plasmids; for example, octopine-type Ti plasmids ...
... is the transfer of antibiotic resistance genes which are carried on plasmids. The plasmids can be ... Other smaller plasmids (usually < 10 kb in size) can be mobilized by a conjugative plasmid (usually > 30 kb) in order to be ... Study investigating physiological effect of pHK01 plasmid in host E.coli J53 found that the plasmid reduced bacterial motility ... are common for resistance plasmids in Enterobacteriaceae. Often multiple beta-lactamase genes are found on the same plasmid ...
Each plasmid has its independent replication system which controls the number of copies of the plasmid in a cell. The higher ... pSK1 is a plasmid from Staphylococcus aureus. This plasmid has a partition system determined by a single gene, par, previously ... For instance, if there are 2 copies of a plasmid in a cell, there is a 50% chance of having one plasmid-less daughter cell. ... A plasmid partition system is a mechanism that ensures the stable inheritance of plasmids during bacterial cell division. ...
When the plasmid concentration is high, RepA plasmids bound to iterons form dimers in between two plasmids, "handcuffing" them ... Low copy number plasmids, also called stringent plasmids, require tighter control of replication. In ColE1 derived plasmids, ... of the plasmid). This is particularly likely with low copy number plasmids. Plasmids can also be incompatible due to shared ... In R1 plasmids RepA can be transcribed from two different promoters. It is made from the first promoter until the plasmid ...
The Bacillus-plasmid RNA motif is a predicted conserved RNA structure usually located in plasmids. It is known in species under ... The fact that the RNA structure is typically found in plasmids suggests that it might be involved in the regulation plasmid ... However, the Bacillus-plasmid RNA motif does not appear to be homologous to any such known RNA. Weinberg Z, Wang JX, Bogue J, ... Page for Bacillus-plasmid RNA at Rfam v t e (Orphaned articles from December 2021, All orphaned articles, Non-coding RNA, All ...
However, many of the plasmids that carry lactis-plasmid RNAs also carry ctRNA-pND324 RNAs, which are involved in plasmid copy ... Therefore lactis-plasmid RNAs might have a different function. R1162-like plasmid antisense RNA Weinberg Z, Wang JX, Bogue J, ... They typically lie near to repB genes, and are almost found in plasmids. This data suggested that lactis-plasmid RNAs ... The lactis-plasmid RNA motif is a conserved RNA structure identified by bioinformatics. The RNAs are restricted to lactic acid ...
... of cardiomyocytes is being examined as a potential treatment for coronary artery disease (a major ... Transfection with HGF plasmids in damaged cardiac tissue also promotes angiogenesis (increased capillary density compared to ... Animal research has demonstrated that administration of HGF cDNA plasmids into ischemic cardiac tissue can increase cardiac ... "Enhanced cardioprotective effects by coexpression of two isoforms of hepatocyte growth factor from naked plasmid DNA in a rat ...
... is a 75-base RNA molecule which negatively regulates the RepI region of the plasmid. The ... "Copy number of the broad host-range plasmid R1162 is determined by the amounts of essential plasmid-encoded proteins". Journal ... Kim K, Meyer RJ (October 1986). "Copy-number of broad host-range plasmid R1162 is regulated by a small RNA". Nucleic Acids ... Page for R1162-like plasmid antisense RNA at Rfam v t e (Antisense RNA, All stub articles, Molecular and cellular biology stubs ...
Smaller independent pieces of DNA, called plasmids, are also found in archaea. Plasmids may be transferred between cells by ... Sota M, Top EM (2008). "Horizontal Gene Transfer Mediated by Plasmids". Plasmids: Current Research and Future Trends. Caister ... ISBN 978-1-904455-27-1. Lipps G (2008). "Archaeal Plasmids". Plasmids: Current Research and Future Trends. Caister Academic ... Schleper C, Holz I, Janekovic D, Murphy J, Zillig W (August 1995). "A multicopy plasmid of the extremely thermophilic archaeon ...
Plasmids containing the Factor IX gene, along with plasmids with a gene that codes for resistance to methotrexate, were ... The recombinant plasmids were inserted into Escherichia coli bacteria, which were "induced to produce 100,000 molecules of ... The gene that was inserted into the plasmid was created by reverse transcription of the mRNA found in pituitary glands to ... The artificial genes were "then inserted... into plasmids... among a group of genes that" are activated by lactose. Thus, the ...
"ABOUT THE WRITER". GENTLE PLASMIDS. JY Yang. 12 January 2017. Retrieved 20 February 2018. Yarian, Karekin (October 11, 2017). " ...
It is used to increase the transfection efficiency of RNA (including mRNA and siRNA) or plasmid DNA into in vitro cell cultures ... DNA plasmids. Lipofectamine consists of a 3:1 mixture of DOSPA (2,3‐dioleoyloxy‐N‐ [2(sperminecarboxamido)ethyl]‐N,N‐dimethyl‐1 ...
Cells harboring the plasmid will survive when exposed to the antibiotic, while those that have failed to take up plasmid ... "plasmid / plasmids , Learn Science at Scitable". www.nature.com. Retrieved 2017-12-06. Shizuya H, Birren B, Kim UJ, Mancino V, ... and a plasmid cloning vector. E. coli and plasmid vectors are in common use because they are technically sophisticated, ... That is, these plasmids could serve as cloning vectors to carry genes. Virtually any DNA sequence can be cloned and amplified, ...
Fertility plasmids, or f plasmids, allow for conjugation to occur whereas resistance plasmids, or r plasmids, contain genes ... Circular bacterial plasmids are classified according to the special functions that the genes encoded on the plasmid provide. ... Circular bacterial plasmids are also the basis for the production of DNA vaccines. Plasmid DNA vaccines are genetically ... The linear plasmids with a covalently attached protein may assist with bacterial conjugation and integration of the plasmids ...
Plasmids in Bacteria. Boston, MA: Springer US. ISBN 1-4613-2447-5. {{cite book}}: ,first1= has generic name (help) Roberts, ...
The G+C base content of the DNA is 46.6%. T. petrophila is also known to contain one of the smallest plasmids. Thermotoga ... "Mobility of plasmids". Microbiology and Molecular Biology Reviews. 74 (3): 434-452. doi:10.1128/MMBR.00020-10. PMC 2937521. ... petrophila RKU1 plasmid (pRKU1) is negatively supercoiled, contains 846 base pairs, and carries only the rep gene. Due to T. ...
As of 2014 Addgene's repository comprised 30,000 plasmids, deposited by 1,700 labs. Its plasmid collection contains plasmids ... Plasmid Cloning Guides Molecular Cloning Guides-References to help scientists design plasmid cloning experiments, including ... The plasmids are available to both academic and industry labs. Noteworthy depositors include:[citation needed] 12 Nobel Prize ... Addgene accepts plasmids from researchers for distribution and archival. Addgene obtains revenues and licensing fees using the ...
Plasmids in Bacteria. Boston, MA: Springer US. ISBN 978-1-4613-2447-8. {{cite book}}: ,first1= has generic name (help) I.W. ...
Kandavelou K, Chandrasegaran S (2008). "Plasmids for Gene Therapy". In Lipps, Georg (ed.). Plasmids: Current Research and ...
Kandavelou K, Chandrasegaran S (2008). "Plasmids for Gene Therapy". Plasmids: Current Research and Future Trends. Norfolk: ... Since ZFN-encoding plasmids could be used to transiently express ZFNs to target a DSB to a specific gene locus in human cells, ... The ZFN-encoding plasmid-based approach has the potential to circumvent all the problems associated with the viral delivery of ... The construction of plasmid vectors is simple and straightforward. Custom-designed ZFNs that combine the non-specific cleavage ...
The two plasmids are pTiC58, responsible for the processes involved in virulence, and pAtC58, once dubbed the "cryptic" plasmid ... To be virulent, the bacterium contains tumour-inducing plasmid (Ti plasmid or pTi), of 200 kbp, which contains the T-DNA and ... PMID 32915125.{{cite journal}}: CS1 maint: uses authors parameter (link) "At plasmid" when talking about related plasmids Smith ... Agrobacterium tumefaciens infects the plant through its Ti plasmid. The Ti plasmid integrates a segment of its DNA, known as T- ...
"Geraldine Seydoux Lab Plasmids". Addgene. (Articles with short description, Short description matches Wikidata, Articles with ...
Maia M.; Sanchez J. M.; Vela G. R. (1988). "Plasmids of Azotobacter vinelandii". Journal of Bacteriology. 170 (4): 1984-1985. ... In addition to chromosomal DNA, Azotobacter can contain plasmids. Azotobacter species are ubiquitous in neutral and weakly ...
The P4 genome can also exist on its own within the host cell and can replicate as a free plasmid. Pruss, G; Goldstein, RN; ... Briani, Federica; Dehò, Gianni; Forti, Francesca; Ghisotti, Daniela (2002). "The Plasmid Status of Satellite Bacteriophage P4 ... ". Plasmid. 45 (1): 1-17. doi:10.1006/plas.2000.1497. PMID 11319927. Claverie JM, Abergel C (2009). "Mimivirus and its ...
... kiviraum/plasmid/plasmid.html , , ( plasp102.zip (239k). ) , , , Authors , , Plasmid Processor program was created during a ... Plasmid Processor is freeware, so you can use and distribute it freely. , , You can download full executable version of Plasmid ... Plasmid Processor is a simple tool for plasmid presentation for scientific and , educational purposes. It features both ... For further information please contact the authors by e-mail, plasmid at uku.fi. , Any comment on program is greatly ...
TetM-containing plasmids may transfer both itself and beta-lactamase plasmids to Neisseria and related species (7). This ... Plasmid-Mediated Antimicrobial Resistance in Neisseria gonorrhoeae -- United States, 1988 and 1989 Because the prevalence of ... An MIC of greater than or equal to 16.0 ug tetracycline/mL identified presumptively an isolate as having plasmid-mediated, high ... Plasmid-mediated resistance to ceftriaxone or spectinomycin has not been observed in N. gonorrhoeae. ...
NEB offers a vast selection DNA plasmids for use in cloning, protein expression, gene expression, and cellular analysis. ... Home Reagents and Molecular Biology Products DNA Plasmid & Substrates Products DNA Plasmids & Substrates. NEB offers a ... DNA Plasmids & Substrates. Close Lambda DNA Close Lambda DNA (dam-) Close M13mp18 RF I DNA Close M13mp18 Single-stranded DNA ... selection of common and specialized DNA plasmids for use in cloning experiments and applications such as protein expression, ...
Redasoft Plasmid allows you to draw plasmids, vector maps and constructs quickly and easily. Circular and linear maps can be ... Need to draw plasmids? vectors? constructs?. Danny Reda reda at globalserve.net Fri Mar 26 16:27:37 EST 1999 *Previous message ... Plasmid has many useful and time-saving features. Learn more about it at http://www.redasoft.com. *Previous message: ...
Pluripotency via plasmids. Researchers are one step closer to reprogramming stem cells that are safe for use in the clinic with ... but his approach involved laborious and repeated insertions with plasmid vectors, and it was only shown to work in mouse cells ... stemcells.wisc.edu/faculty/thomson.html of the University of Wisconsin-Madison has extended the plasmid technique to reprogram ...
... and gigaprep sizes for small to large scale purification of plasmid DNA for molecular cloning, transformation, and recombinant ... Plasmid DNA purification kits offered in miniprep, midiprep, maxiprep, ... GenElute™ HP plasmid prep kits. GenElute™ HP plasmid purification kits yield high quality plasmid DNA in less than 30 minutes ... GenElute™ plasmid miniprep kits offer simple, rapid, and cost-effective methods for isolating plasmid DNA from E. coli cultures ...
ORF of cylicin, basic protein of sperm head cytoskeleton 2 (CYLC2) in pENTER vector with CMV promoter and C-terminal FLAG and His tags.
Explore the breadth of our plasmid prep offerings to discover the right solution for your needs. ... We offer a range of kits to extract plasmid DNA at several scales and by multiple processing methods, including completely ... Plasmid Purification Compare Plasmid Purification Kits. We offer a range of kits to extract plasmid DNA at several scales and ... PureYield™ Plasmid Miniprep System. 600ul to 3ml. 10 minutes. Fully automated purification of transfection-grade plasmid, with ...
Human ORAI1 cDNA Clone BC013386 pENTR223.1 or pUC Plasmid from Cusabio. Cat Number: CSB-CL846601HU2. USA, UK & Europe ... Cusabio Customized Plasmid Human ORAI1 cDNA Clone BC013386 pENTR223.1 or pUC Plasmid. (No reviews yet) Write a Review Write a ... 10 μg plasmid + 200μl Glycerol. Gene:. ORAI1. Gene ID:. 84876. Accession #:. BC013386. Plasmid:. Vector will be determined ... Cusabio Customized Plasmid. Human ORAI1 cDNA Clone BC013386 pENTR223.1 or pUC Plasmid. ...
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Plasmids containing this gene, or a homologous gene.. ID. Plasmid. Gene/Insert. PI. ... Addgene is a nonprofit plasmid repository.. We store 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 ... How can I be notified when a plasmid from a specific lab or paper is available? ...
Beckman Coulter Life Sciences offers EMnetik Plasmid Purification Kit, a revolutionary bead-based DNA cleanup kit using the ... Benefits of the EMnetik Plasmid Prep System:. *Plasmid recovery of 4-7 µg shown for a high copy plasmid ... EMnetik Plasmid Purification Kit, C68445. Product No:C68445. DNA Cleanup for Genetic Engineering. The EMnetik Plasmid ... a user can complete a plasmid prep without having to handle columns. ...
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Other sequences of Citrobacter freundii ...
Therefore, Platon analyzes the natural distribution biases of certain protein coding genes between chromosomes and plasmids. ... Platon detects plasmid contigs within bacterial draft genomes from WGS short-read assemblies. ... Platon - identification and characterization of bacterial plasmid contigs. Platon detects plasmid contigs within bacterial ... Platon - identification and characterization of bacterial plasmid contigs Platon detects plasmid contigs within bacterial draft ...
... which is constituted by pairs of plasmids, generically indicated as pSiMPl,sup,x,/sup,_N and pSiMPl,sup,x,/sup,_C, which can be ... We recently developed the SiMPl plasmid toolbox, which is constituted by pairs of plasmids, generically indicated as pSiMPlx_N ... whereby each enzyme fragment is expressed from one of the plasmids in the pair. pSiMPl plasmids are currently available for use ... Expanding the SiMPl Plasmid Toolbox for Use with Spectinomycin/Streptomycin ACS Omega. 2021 May 21;6(22):14148-14153. doi: ...
Eukaryotic Plasmids (1 Lecture) The yeast plasmid: a highly persistent selfish DNA element The yeast plasmid: a highly ... Introduction and General Properties of Plasmids (2 Lectures) Introduction to plasmid biology Introduction to plasmid biology ... Evolution of plasmids and their role in bacterial diversity… Evolution of plasmids and their role in bacterial diversity and ... Plasmids, integrons and the spread of antibiotic resistance Plasmids, integrons and the spread of antibiotic resistance ...
Analytical PCR experiments ideally use inside probes for monitoring the amplification response and particular detection of the amplicon. Such inside probes must be designed in shut context with the amplification primers, and will require further issues for the detection of genetic variations. Right here we describe Edesign, a brand new on-line and stand-alone software for […]. Continue Reading ...
Add 0.1 volume of 3 M sodium acetate and 3 volume of ethanol into the plasmid solution. Spin at 14,000 rpm for 5 min at 4 C. ... 1. Shake E. coli harboring plasmid for 16-24 hr at 37 C in 3.3 ml of TB containing appropriate antibiotics. (when using ... Extract the plasmid solution with chloroform to remove PEG. Take aquaous phase. Extract the aquaous phase with phenol. Take ... This improves the yield of plasmids.). 2. Collect the bacterial cells in a microfugte tube by spinning for 1 min at 10,000 rpm ...
Shehabi, A.A., Mahafzah, A.M. & Al Khalili, K.Z. (‎2004)‎. Antimicrobial resistance and plasmid profiles of urinary Escherichia ... Antimicrobial resistance and plasmid profiles of urinary Escherichia coli isolates from Jordanian patients. ... We investigated antimicrobial resistance patterns and plasmid profiles of uropathogenic Escherichia coli isolates from ... transferable R-plasmid of 28 kb was found in most E. coli isolates [‎67%]‎ that were resistant to at least ampicillin, ...
Fc-fusion protein expression plasmid - Engineered human IgG2, reduced CDC pFUSE-hIgG4e1-Fc Fc-fusion protein expression plasmid ... pFUSE-hIgG1e1-Fc Fc-fusion protein expression plasmid - Engineered human IgG1, increased half-life pFUSE-hIgG1e2-Fc Fc-fusion ... pFUSE-mIgG1e2-Fc Fc-fusion protein expression plasmid - Engineered murine IgG1, increased protein A affinity pFUSE-mIgG2Ae1-Fc ... protein expression plasmid - Engineered human IgG1, increased half-life pFUSE-hIgG1e4-Fc Fc-fusion protein expression plasmid ...
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R391: a conjugative integrating mosaic comprised of phage, plasmid, and transposon elements. ...
Our platform approach to GMP Plasmid DNA production minimizes the time needed from project set-up up to Drug Substance or Drug ... Plasmid QC Test Package Example. The QC tests for plasmids include e.g. the following:. Plasmid QA Documentation Package ... Production of AAV plasmids. Three separate E. coli processes are needed to produce the necessary AAV plasmids for transfection ... Then the AAV plasmids are purified with chromatographic methods. Finally, the plasmids will be analyzed e.g. for their identity ...
Plasmid Kits collection, the ZymoPURE™ Plasmid Miniprep Kit features a spin column-based method for the purification of up to ... 100 µg of ultra-pure endotoxin-free plasmid DNA in less than 15 minutes. The unique spin-column design also provides zero ... Part of the ZymoPURE™ Plasmid Kits collection, the ZymoPURE™ Plasmid Miniprep Kit features a spin column-based method for the ... Q7: Is there a protocol for low-copy number plasmid DNA? Yes, the low-copy protocol can be found in the Appendix of the kit ...
Zhong Z, Caspi R, Mincer T, Helinski D, Knauf V, Boardman K, Wilkinson JE, Shea T, DeLoughery C, Toukdarian A. A 50-kb plasmid ... A 50,709-bp cryptic plasmid isolated from a marine Micrococcus has been sequenced and found to contain a number of putative ... A 50-kb plasmid rich in mobile gene sequences isolated from a marine micrococcus. ... The coding regions for 11 putative transposases comprise approximately 17% of the total plasmid sequence. The majority of these ...
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Paav Plasmid, packaging plasmid, Packaging Vector, Pacyc Plasmid, Pacyc177, Pam Sequence Crispr, Pax2 Antibody, Pbad, Pbad ... Pbs Plasmid, Pcag Gfp, Pcaggs Vector, Pcdna, pcdna 3, Pcdna 3.1, pcdna 3.4, pcdna gfp, pcdna map, Pcdna Plasmid, Pcdna Vector, ... Pbs Plasmid The role of Colchicine on actin polymerization dynamics: as a potent anti-angiogenic factor. November 2, 2022. ... Tags Nfkb Reporter Plasmid, nfyb, Nickase, Nickase Cas9, Novagen Pet Vectors, Oligo Primer Design, Online Primer Design, order ...
  • NEB offers a selection of common and specialized DNA plasmids for use in cloning experiments and applications such as protein expression, gene expression, and cellular analysis. (neb.com)
  • Receive email alerts when new plasmids with this gene become available. (addgene.org)
  • Plasmids containing this gene, or a homologous gene. (addgene.org)
  • In addition, plasmids have been very much involved as important tools in basic research in molecular biology (cloning, sequencing and genetic manipulation) as well as the biotechnology associated with genetic engineering and gene therapy. (hstalks.com)
  • Plasmids can be searched by gene name, symbol or ID on our gRNA Database. (genscript.com)
  • One of our main focus areas is viral vector manufacturing for those gene therapies, where plasmid DNA constructs serve as key raw material. (biovian.com)
  • Our GMP plasmid DNA may also be used to develop novel DNA or mRNA vaccines and non-viral gene therapy applications. (biovian.com)
  • A 50-kb plasmid rich in mobile gene sequences isolated from a marine micrococcus. (sri.com)
  • marine sediment bacterium Micrococcus plasmid DNA sequence analysis transposase gene cluster. (sri.com)
  • Cytovance Biologics , a biopharmaceutical contract development and manufacturing organization (CDMO) of mammalian, microbial biologics and gene therapy plasmid DNA (pDNA), has announced a licensing opportunity for pDNA manufacturing. (contractpharma.com)
  • The companies have signed an agreement for the production of two plasmids needed for the manufacture of CG01, a gene therapy for the treatment of drug-resistant focal epilepsy. (pharmtech.com)
  • Cobra Biologics, a contract development and manufacturing organization (CDMO), announced on Sept. 10, 2020 that it has signed an agreement with CombiGene, a gene therapy company located in Sweden, for the good manufacturing practice (GMP) production of two plasmids needed for the manufacture of CG01, a gene therapy for the treatment of drug-resistant focal epilepsy. (pharmtech.com)
  • The replication of plasmid and phage DNA's could be controlled independently by a mutation in the host bacterium and mutations in T₇ gene 5 (DNA polymerase), respectively. (ubc.ca)
  • A portion of this plasmid was derived from a plasmid made by eGFP gene was synthetically produced. (addgene.org)
  • 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)
  • Genetic environments of the qnrS2 gene in plasmid p37 from Aeromonas punctata 37 and comparison with related plasmid structures. (cdc.gov)
  • Hata M , Suzuki M , Matsumoto M , Takahashi M , Sato K , Ibe S , Cloning of a novel gene for quinolone resistance from a transferable plasmid in Shigella flexneri 2b. (cdc.gov)
  • Jacoby GA , Walsh KE , Mills DM , Walker VJ , Oh H , Robicsek A , qnrB , another plasmid-mediated gene for quinolone resistance. (cdc.gov)
  • High-efficiency in vivo gene transfer using intraarterial plasmid DNA injection following in vivo electroporation. (nature.com)
  • The carriers were complexed with episomal plasmid DNA or minicircles using secreted alkaline phosphatase (SEAP) gene as a marker gene. (helsinki.fi)
  • According to the results, PBuA-PDMAEMA-polymers and DOTAP/DOPE/PS-liposomes complexed with episomal plasmid or minicircles are potential gene delivery agents for further studies in AMD. (helsinki.fi)
  • The mcr-1 gene exists on a plasmid, a small piece of DNA that is capable of moving from one bacterium to another, potentially spreading antibiotic resistance to other bacterial species. (cdc.gov)
  • In November 2015, a report from China first described plasmid-mediated colistin-resistance caused by the mcr-1 gene. (cdc.gov)
  • The presence of the mcr-1 gene on a plasmid means that colistin resistance can be shared with other more resistant bacteria such as CRE, raising the possibility that untreatable bacteria could develop. (cdc.gov)
  • 20 kb) plasmids containing cloned gene clusters. (cdc.gov)
  • The bla NDM-1 gene was encoded on plasmids that were easily transferable. (who.int)
  • Editorial Note: Although national gonorrhea rates changed little from 1988 (302 per 100,000 persons) to 1989 (298 per 100,000), important increases occurred in the percentage of isolates with plasmid-mediated resistance. (cdc.gov)
  • We investigated antimicrobial resistance patterns and plasmid profiles of uropathogenic Escherichia coli isolates from inpatients and outpatients at Jordan University Hospital in 2000 and 2001. (who.int)
  • The plasmid content of ESBL producing isolates and their participation in drug resistance were investigated. (scirp.org)
  • 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)
  • The plasmid spread to unrelated E. coli isolates within an index cattle farm and persisted within the environment. (cdc.gov)
  • In this study, we report the full sequence and analysis of pCT and demonstrate the spread of pCT-like plasmids in animal and human E. coli isolates from the United Kingdom, Europe, Australia, and Asia. (cdc.gov)
  • Cattoir V , Poirel L , Rotimi V , Soussy CJ , Nordmann P . Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates. (cdc.gov)
  • We studied the characteristics of virulence plasmid using serological, biochemical and bioassay tests in Y. enterocolitica isolates of chicken using plasmid curing. (who.int)
  • Total Plasmid Chromosomal CLINIC Isolates Resistance No. % Resistance No. (cdc.gov)
  • The outbreak isolates were characterized by antimicrobial drug resistance and plasmid and pulsed-field gel electrophoresis profiles. (cdc.gov)
  • Plasmid analyses were performed on beta-lactamase-producing isolates. (bvsalud.org)
  • Part of the ZymoPURE™ Plasmid Kits collection, the ZymoPURE™ Plasmid Miniprep Kit features a spin column-based method for the purification of up to 100 µg of ultra-pure endotoxin-free plasmid DNA in less than 15 minutes. (zymoresearch.com)
  • The ZymoPURE - Express Plasmid Midiprep Kit utilizes a patented alkaline lysis method for purifying up to 1.2 mg of high-quality endotoxin-free plasmid DNA directly from the culture without pelleting and resuspension steps. (thomassci.com)
  • We recently developed the SiMPl plasmid toolbox, which is constituted by pairs of plasmids, generically indicated as pSiMPl x _N and pSiMPl x _C, which can be stably maintained in Escherichia coli with a single antibiotic x. (nih.gov)
  • Escherichia coli plasmid pB15 contains genes for resistance to several antibiotics, including tetracycline. (datadryad.org)
  • Two of the seven plasmid-bearing strains were resistant to chloramphenicol (Cm) and tetracycline (Tc) and they transferred Cm and Tc resistance traits to Escherichia coli K12 at frequencies from 1.6 X 10 7 to 1.9 x 10 6 . (monash.edu)
  • This paper studies the potential usefulness of nonlinear regression methods for predicting, from in situ near-infrared (NIR) and mid-infrared (MIR) spectra acquired in high-throughput mode, biomass and plasmid concentrations in Escherichia coli DH5-α cultures producing the plasmid model pVAX-LacZ. (it.pt)
  • Interaction of the plasmid-encoded quinolone resistance protein Qnr with Escherichia coli DNA gyrase. (cdc.gov)
  • Our plasmid kits provide reliable isolation of high quality plasmid DNA. (sigmaaldrich.com)
  • GenElute™ HP plasmid purification kits yield high quality plasmid DNA in less than 30 minutes for Mini, Midi, and Maxiprep kits, or 1.5 hours for Mega and Gigaprep kits. (sigmaaldrich.com)
  • GenElute™ five-minute miniprep kits feature a streamlined protocol yielding up to 5 mg high-quality plasmid DNA in about five minutes. (sigmaaldrich.com)
  • ZymoPURE II Plasmid Isolation kits provide the fastest and simplest method available to efficiently isolate transfection quality plasmid DNA from E. coli. (thomassci.com)
  • We have a well-established plasmid production platform and in-house expertise in quality control that will ensure the delivery of GMP quality plasmid. (pharmtech.com)
  • Conjugal transfer of plasmid-borne multiple antibiotic resistance in Streptococcus faecalis var. (wikidata.org)
  • This situation causes plasmids and bacteria to sometimes experience differing selection pressures. (datadryad.org)
  • 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)
  • The ability and frequency with which antimicrobial resistance genes disseminate between bacteria in humans, the environment, and animals is still debated, and the role of plasmids in this movement between ecosystems, including the food chain, is also still contested, despite mounting evidence that it occurs ( 8,9 ). (cdc.gov)
  • Bacteria occasionally carry DNA in smaller rings known as plasmids. (si.edu)
  • Last year, Kyoto University's linkurl:Shinya Yamanaka;http://www.med.kyoto-u.ac.jp/E/grad_school/introduction/1517/ devised the linkurl:first virus-free route;http://www.sciencemag.org/cgi/content/full/322/5903/949 to obtaining iPS cells, but his approach involved laborious and repeated insertions with plasmid vectors, and it was only shown to work in mouse cells. (the-scientist.com)
  • By characterizing commensal E. coli from Shigella-infected and healthy children, we identified an extensive array of AMR genes and plasmids, including an identical MDR plasmid isolated from both S. sonnei and E. coli in the gut of a single child. (cam.ac.uk)
  • In addition to pathogens, the gut microbiome can house a wide variety of AR genes and plasmids. (cdc.gov)
  • Here, we consider that the plasmid-borne origin-of-transfer substrates encode specific DNA structural properties that can facilitate finding these regions in large datasets, and develop a DNA structure-based alignment procedure for typing the transfer substrates that outperforms mere sequence-based approaches. (biorxiv.org)
  • In this study, approximately 2% of C. perfringens strains, representing the principal biotypes, were found to harbor the urease structural genes, ureABC, and these were localized on large plasmids that often encode, in addition, the lethal epsilon or iota toxins or the enterotoxin. (pasteur.fr)
  • I have a plasmid i want to make and inster in yeast but there are genes to encode for multiple protiens. (sciencemadness.org)
  • Hereby, Platon tries to circularize the contig sequences, searches for rRNA, replication, mobilization and conjugation genes, oriT sequences, incompatibility group DNA probes and finally performs a BLAST+ search against the NCBI plasmid database. (uni-giessen.de)
  • Antimicrobial resistance poses a great danger to humanity, in part due to the widespread horizontal transfer of plasmids via conjugation. (biorxiv.org)
  • Preliminary sequence analyses suggested that divergence in plasmid conjugation was due to altered configurations of a shufflon region (a site-specific recombination system), where genetic rearrangements affect conjugative ability. (datadryad.org)
  • Often a single plasmid is transmitted from one bacterium to another in a sex-like interaction called conjugation. (si.edu)
  • IMSEAR at SEARO: Drug resistant plasmids of Pseudomonas aeruginosa. (who.int)
  • Pai SR, Joshi L. Drug resistant plasmids of Pseudomonas aeruginosa. (who.int)
  • As an added convenience, the ZymoPURE™ Plasmid Miniprep Kit contains colored buffers that permit error-free visualization and identification of complete bacterial cell lysis and neutralization. (zymoresearch.com)
  • The Zyppy Plasmid Miniprep Kit features a pellet-free modified alkaline lysis method that bypasses bacterial culture centrifugation and resuspension steps common to classical plasmid preparation procedures. (thomassci.com)
  • Up to 10 mg (giga), 2.5 mg (mega), 500 µg (maxi), 100 µg (midi), and 20 µg (mini) high-copy plasmid DNA is purified from culture (culture volumes depend on plasmid copy number, size of insert, host strain, and culture medium). (qiagen.com)
  • Q9: Can the ZymoPURE kits be used to isolate large plasmid constructs (BAC/PAC)? (zymoresearch.com)
  • Seven (6.1%) of 115 strains of Salmonella typhi isolated from Malaysian patients harbored a single large plasmid of 71 to 166 mD. (monash.edu)
  • CMV-luciferase was used as a reporter plasmid. (cdc.gov)
  • In an attempt to understand the molecular basis of carcinogenesis induced by these particles, we investigated the potential activation of activator protein-1 (AP-1) by crocidolite and freshly fractured or aged crystalline silica in a JB6 P + cell line stably transfected with AP-1-luciferase reporter plasmid (in vitro) and in AP-1-luciferase reporter transgenic mice (in vivo). (cdc.gov)
  • therefore, dissemination has been attributed to conjugative plasmids rather than to clonal expansion of a bacterial host strain ( 13 ). (cdc.gov)
  • Effects of ingested (i) pediocin PA-1 producing Lactobacillus plantarum DDEN 11007, (ii) the plasmid cured pediocin negative L. plantarum DDEN 12305, or (iii) supernatants of either of these two strains on the composition of the intestinal microbiota of Human Microbiota Associated (HMA) rats were examined by selective cultivation and molecular methods. (dtu.dk)
  • Objective: To isolate ESBLs producing uropathogens and the plasmid underlying their resistance to antibiotics. (scirp.org)
  • described an ESBL-producing isolate from calves with diarrhea that carried bla CTX-M-14 on an IncK plasmid, denoted pCT ( 15 , 16 ). (cdc.gov)
  • plasmids pGNB2 and pMG308 are from a wastewater treatment plant from Germany (unknown bacterial reservoir) ( 24 ) and from a non-Typhi Salmonella clinical isolate from the United States ( 25 ), respectively. (cdc.gov)
  • Exact values for these thresholds have been computed based on Monte Carlo simulations of artifical replicon fragments created from complete RefSeq chromosome and plasmid sequences. (uni-giessen.de)
  • GenScript maintains a collection of more than 20,000 lentiCRISPRv2 plasmids containing guide RNA (gRNA) sequences pre-validated by the Broad Institute. (genscript.com)
  • Roche Genopure™ midi and maxi prep kits use anion exchange chromatography for plasmid purification. (sigmaaldrich.com)
  • The lysate can be filtered rather than centrifuged prior to anion exchange, resulting in shorter plasmid prep times and complete removal of SDS from the purified product. (sigmaaldrich.com)
  • QIAGEN Plasmid Kits provide gravity-flow, anion-exchange tips for purification of transfection-grade plasmid DNA. (qiagen.com)
  • The QIAGEN Plasmid Kits uses gravity-flow QIAGEN anion-exchange tips for efficient purification of plasmid DNA. (qiagen.com)
  • Cattoir V , Poirel L , Mazel D , Soussy CJ , Nordmann P . Vibrio splendidus as the source of plasmid-mediated QnrS-like quinolone resistance determinants. (cdc.gov)
  • The widespread presence of a family of fish virulence plasmids in Vibrio vulnificus stresses its relevance as a zoonotic pathogen linked to fish farms. (bvsalud.org)
  • Martinez-Martinez L , Pascual A , Jacoby GA . Quinolone resistance from a transferable plasmid. (cdc.gov)
  • Plasmid purification kits are available based on size of the bacterial culture and corresponding plasmid yield (miniprep, midiprep, maxiprep, megaprep, and gigaprep). (sigmaaldrich.com)
  • The P2 plasmid from the Zika virus was converted into cDNA and injected into mouse models. (unl.edu)
  • ZymoPURE™ technology uses a modified alkaline lysis method and features novel binding chemistry that yields highly concentrated plasmid DNA (up to 3 µg/µl). (zymoresearch.com)
  • Hi, does anyone know the maximum capacity for a plasmid that yeast could recive. (sciencemadness.org)
  • 10 kbp plasmids are fine for yeast (just make sure you have all the proper elements, like a yeast origin of replication and a selectable marker). (sciencemadness.org)
  • In fact, yeast can have plasmids much larger than this. (sciencemadness.org)
  • A plasmid that is maintained needs to have a yeast (and probably a bacterial) origin of replication. (sciencemadness.org)
  • Further, we associate these clades, most of them previously but incompletely described, with the acquisition of a family of fish virulence plasmids containing genes essential for resistance to the immune system of certain teleosts of interest in aquaculture . (bvsalud.org)
  • Pathogenic Yersinia enterocolitica harbour plasmid that is essential for virulence. (who.int)
  • Unexpected Occurrence of Plasmid-Mediated Quinolone Resistance Determinants in Environmental Aeromonas spp. (cdc.gov)
  • Polymerase chain reaction (PCR) assays and sequencing was used to determine the presence of β-lactamase encoding genes (bla) including bla NDM-1 and plasmid-mediated quinolone and aminoglycoside resistance determinants. (who.int)
  • Redasoft Plasmid allows you to draw plasmids, vector maps and constructs quickly and easily. (bio.net)
  • CsCI isolation is a widely used method for isolating highly pure circular plasmid DNA in relatively high yield from bacterial cells. (abnova.com)
  • Alkaline lysis is one of the most generally useful methods for isolating circular plasmid DNA from bacterial cells. (abnova.com)
  • Plasmid purification is commonly used for molecular cloning, transformation, and recombinant protein expression. (sigmaaldrich.com)
  • Recombinant plasmid pAS37 has been obtained from our study. (cdc.gov)
  • These scores express the empirically measured frequency biases of protein sequence distributions between plasmids and chromosomes pre-computed on complete NCBI RefSeq replicons. (uni-giessen.de)
  • These plasmids contain a 17bp-1.8kb expressible linker in lieu of a customized sgRNA sequence, which can be modified by your laboratory. (genscript.com)
  • The coding regions for 11 putative transposases comprise approximately 17% of the total plasmid sequence. (sri.com)
  • Also, in plasmids I have seen premade, they seem to have random dna inbetween each sequence. (sciencemadness.org)
  • Plasmid sequence and annotations. (addgene.org)
  • Use text editor or plasmid mapping software to view sequence. (addgene.org)
  • In 2004, an ESBL- carrying IncK plasmid (pCT) was isolated from cattle in the United Kingdom. (cdc.gov)
  • Our offerings include the latest CRISPR plasmids and databases developed by the CRISPR pioneering Feng Zhang laboratory. (genscript.com)
  • Broad Institute-validated plasmids are a well-tested platform for expressing CRISPR/Cas9, and avoid instability issues in RNA-based platforms. (genscript.com)
  • A variety of plasmids and pooled guide RNA libraries for CRISPR-mediated genome manipulation generated by IGI member labs and others can be acquired through Addgene . (innovativegenomics.org)
  • Martinez-Martinez L , Pascual A , Garcia I , Tran J , Jacoby GA . Interaction of plasmid and host quinolone resistance. (cdc.gov)
  • The worldwide emergence of plasmid-mediated quinolone resistance. (cdc.gov)
  • Poirel L , Rodriguez-Martinez JM , Mammeri H , Liard A , Nordmann P . Origin of plasmid-mediated quinolone resistance determinant QnrA. (cdc.gov)
  • Therefore, Platon analyzes the natural distribution biases of certain protein coding genes between chromosomes and plasmids. (uni-giessen.de)
  • Instead, their DNA can be found in the cytoplasm in a region called the nucleoid or in circular chromosomes called plasmids. (visiblebody.com)
  • A 50,709-bp cryptic plasmid isolated from a marine Micrococcus has been sequenced and found to contain a number of putative mobile genetic elements. (sri.com)
  • Marker rescue between a plasmid carrying T₇⁺ DNA and a mutant bacteriophage was used to study the role of replication in genetic recombination. (ubc.ca)
  • Results indicated that when only one molecule, i.e. plasmid or phage, could replicate, recombination was decreased slightly. (ubc.ca)
  • Density transfer experiments examined recombination between 3 a heavy labeled phage and a H-labeled plasmid molecule. (ubc.ca)
  • A model for early events during plasmid-phage recombination is presented. (ubc.ca)
  • Megaprep plasmid DNA extraction kit is used when the starting E. coli culture volume is 500 ml~2.5 L of LB broth and the expected DNA yield is 1.5~2.5 mg. (abnova.com)
  • This represents the first report of a plasmid-encoded urease in a gram-positive bacterium. (pasteur.fr)
  • We identify thousands of yet undiscovered DNA transfer substrates, showing that actual plasmid mobility can in fact be 2-fold higher and span almost 2-fold more host species than is currently known. (biorxiv.org)
  • Plasmid DNA purified with QIAGEN Plasmid Kits is highly suitable for use in applications such as transfection (see figure " Transfection efficiency versus plasmid purification method. Different pRSVcat DNA preparations using the methods indicated were introduced into the indicated cell lines by liposome-mediated transfection, and the efficiencies determined by measuring CAT expression levels after 40 h. Each bar represents the mean of 4 independent transfections (2 transfections with each of 2 independent plasmid preparations). The highest transfection efficiency was achieved with QIAGEN Plasmid Kits. "> Transfection efficiency versus plasmid purification method "), cloning, and in vitro transcription. (qiagen.com)
  • GFP and pRL-TK plasmids had been utilized to monitor transfection efficiency. (woofahs.com)
  • The ZR Plasmid Miniprep-Classic is designed for efficient isolation of plasmid DNA from E. coli using a traditional 3-buffer (P1, P2, P3) procedure that is simple, rapid, user-friendly, and reliable. (thomassci.com)
  • This improves the yield of plasmids. (kyoto-u.ac.jp)
  • For high copy plasmids using more culture can cause overloading and subsequent clogging of columns, which can result in poor DNA yield and/or quality. (zymoresearch.com)
  • Our platform is uniquely designed to process plasmids with low shear lysis and purification steps to produce high-quality, high-yield pDNA. (contractpharma.com)
  • Midiprep plasmid DNA extraction kit is used when the starting E. coli culture volume is 15-25 ml of LB broth and the expected DNA yield is 100-350 μg. (abnova.com)
  • When plasmid-bearing cells were experimentally evolved in the laboratory, changes in resistance level in the unselected tetracycline marker coincided with changes in plasmid rates of vertical versus horizontal transmission. (datadryad.org)
  • Furthermore, we proposed that correlated resistance and transmission in pB15 derivatives were caused by a tetracycline-resistance transposon inserted into a transfer operon, allowing transcription from its promoter to simultaneously affect both plasmid resistance and transmission. (datadryad.org)
  • Addgene is a nonprofit plasmid repository. (addgene.org)
  • Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications. (addgene.org)
  • Other extraction and purification plasmid DNA Kits are available from varying manufacturers, named by size of bacterial culture, includes gigaprep, midiprep, and miniprep. (abnova.com)
  • In addition, the wash regimen has been optimized to ensure the plasmid DNA is free of endotoxins, salt, protein, and RNA. (zymoresearch.com)
  • We identified several independent acquisitions of XDR /MDR-inducing plasmids, likely facilitated by horizontal transfer from commensals in the human gut. (cam.ac.uk)
  • Because of their relatively small size, plasmids have also served as easily studied models for DNA replication and maintenance while at the same time illustrating great mechanistic diversity. (hstalks.com)
  • Integration plasmids don't need a end host origin of replication, as they have either one or two regions that facilitate cross-over. (sciencemadness.org)
  • I am new to plasmid design and I wanted to know if i needed to put in start and stops for each protein if i want them to come out separately as different protiens. (sciencemadness.org)
  • Our previous study showed that the chlamydial plasmid-encoded protein pGP3 forms a stable complex with LL-37 to neutralize its pro-inflammatory activity. (medscimonit.com)
  • QIAGEN Plasmid Kits are intended for molecular biology applications. (qiagen.com)
  • The aim of these presentations is to describe some of the key properties of plasmids that have made them such important elements in our understanding and exploitation of many aspects of molecular genetics and evolution. (hstalks.com)
  • Engineering of DNA vaccines using molecular adjuvant plasmids. (elsevier.com)
  • Dive into the research topics of 'Engineering of DNA vaccines using molecular adjuvant plasmids. (elsevier.com)