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.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
The formation of one or more genetically identical organisms derived by vegetative reproduction from a single cell. The source nuclear material can be embryo-derived, fetus-derived, or taken from an adult somatic cell.
The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
The functional hereditary units 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.
A large collection of DNA fragments cloned (CLONING, MOLECULAR) from a given organism, tissue, organ, or cell type. It may contain complete genomic sequences (GENOMIC LIBRARY) or complementary DNA sequences, the latter being formed from messenger RNA and lacking intron sequences.
The sequential correspondence of nucleotides in one nucleic acid molecule with those of another nucleic acid molecule. Sequence homology is an indication of the genetic relatedness of different organisms and gene function.
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
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).
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.
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.
Proteins prepared by recombinant DNA technology.
Detection of RNA 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.
Any method used for determining the location of and relative distances between genes on a chromosome.
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
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.
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.
Deoxyribonucleic acid that makes up the genetic material of bacteria.
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.
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.
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).
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.
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)
The relationships of groups of organisms as reflected by their genetic makeup.
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.
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.
Accumulation of a drug or chemical substance in various organs (including those not relevant to its pharmacologic or therapeutic action). This distribution depends on the blood flow or perfusion rate of the organ, the ability of the drug to penetrate organ membranes, tissue specificity, protein binding. The distribution is usually expressed as tissue to plasma ratios.
A form of GENE LIBRARY containing the complete DNA sequences present in the genome of a given organism. It contrasts with a cDNA library which contains only sequences utilized in protein coding (lacking introns).
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.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
A set of genes descended by duplication and variation from some ancestral gene. Such genes may be clustered together on the same chromosome or dispersed on different chromosomes. Examples of multigene families include those that encode the hemoglobins, immunoglobulins, histocompatibility antigens, actins, tubulins, keratins, collagens, heat shock proteins, salivary glue proteins, chorion proteins, cuticle proteins, yolk proteins, and phaseolins, as well as histones, ribosomal RNA, and transfer RNA genes. The latter three are examples of reiterated genes, where hundreds of identical genes are present in a tandem array. (King & Stanfield, A Dictionary of Genetics, 4th ed)
The sum of the weight of all the atoms in a molecule.
Proteins found in any species of bacterium.
Established cell cultures that have the potential to propagate indefinitely.
Characteristic restricted to a particular organ of the body, such as a cell type, metabolic response or expression of a particular protein or antigen.
A multistage process that includes the determination of a sequence (protein, carbohydrate, etc.), its fragmentation and analysis, and the interpretation of the resulting sequence information.
Plasmids containing at least one cos (cohesive-end site) of PHAGE LAMBDA. They are used as cloning vehicles.
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 characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.
Transport proteins that carry specific substances in the blood or across cell membranes.
Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin.
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.
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.
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.
The functional hereditary units of FUNGI.
The degree of similarity between sequences. Studies of AMINO ACID SEQUENCE HOMOLOGY and NUCLEIC ACID SEQUENCE HOMOLOGY provide useful information about the genetic relatedness of genes, gene products, and species.
A technique that localizes specific nucleic acid sequences within intact chromosomes, eukaryotic cells, or bacterial cells through the use of specific nucleic acid-labeled probes.
Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.
Proteins found in plants (flowers, herbs, shrubs, trees, etc.). The concept does not include proteins found in vegetables for which VEGETABLE PROTEINS is available.
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.
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.
A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
A sequence of amino acids in a polypeptide or of nucleotides in DNA or RNA that is similar across multiple species. A known set of conserved sequences is represented by a CONSENSUS SEQUENCE. AMINO ACID MOTIFS are often composed of conserved sequences.
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.
A genus of bacteria that form a nonfragmented aerial mycelium. Many species have been identified with some being pathogenic. This genus is responsible for producing a majority of the ANTI-BACTERIAL AGENTS of practical value.
DNA 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 rate dynamics in chemical or physical systems.
The functional hereditary units of PLANTS.
A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.
The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA.
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.
A process whereby multiple RNA transcripts are generated from a single gene. Alternative splicing involves the splicing together of other possible sets of EXONS during the processing of some, but not all, transcripts of the gene. Thus a particular exon may be connected to any one of several alternative exons to form a mature RNA. The alternative forms of mature MESSENGER RNA produce PROTEIN ISOFORMS in which one part of the isoforms is common while the other parts are different.
A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in enzyme synthesis.
Sequences of DNA in the genes that are located between the EXONS. They are transcribed along with the exons but are removed from the primary gene transcript by RNA SPLICING to leave mature RNA. Some introns code for separate genes.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
Deoxyribonucleic acid that makes up the genetic material of fungi.
Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections.
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.
A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.
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.
Sequences of DNA or RNA that occur in multiple copies. There are several types: INTERSPERSED REPETITIVE SEQUENCES are copies of transposable elements (DNA TRANSPOSABLE ELEMENTS or RETROELEMENTS) dispersed throughout the genome. TERMINAL REPEAT SEQUENCES flank both ends of another sequence, for example, the long terminal repeats (LTRs) on RETROVIRUSES. Variations may be direct repeats, those occurring in the same direction, or inverted repeats, those opposite to each other in direction. TANDEM REPEAT SEQUENCES are copies which lie adjacent to each other, direct or inverted (INVERTED REPEAT SEQUENCES).
CELL LINES derived from the CV-1 cell line by transformation with a replication origin defective mutant of SV40 VIRUS, which codes for wild type large T antigen (ANTIGENS, POLYOMAVIRUS TRANSFORMING). They are used for transfection and cloning. (The CV-1 cell line was derived from the kidney of an adult male African green monkey (CERCOPITHECUS AETHIOPS).)
A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.
Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.
Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.
A group of adenine ribonucleotides in which the phosphate residues of each adenine ribonucleotide act as bridges in forming diester linkages between the ribose moieties.
Methods of implanting a CELL NUCLEUS from a donor cell into an enucleated acceptor cell.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
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.
Deoxyribonucleic acid that makes up the genetic material of plants.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.
A 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 theoretical representative nucleotide or amino acid sequence in which each nucleotide or amino acid is the one which occurs most frequently at that site in the different sequences which occur in nature. The phrase also refers to an actual sequence which approximates the theoretical consensus. A known CONSERVED SEQUENCE set is represented by a consensus sequence. Commonly observed supersecondary protein structures (AMINO ACID MOTIFS) are often formed by conserved sequences.
Mapping of the linear order of genes on a chromosome with units indicating their distances by using methods other than genetic recombination. These methods include nucleotide sequencing, overlapping deletions in polytene chromosomes, and electron micrography of heteroduplex DNA. (From King & Stansfield, A Dictionary of Genetics, 5th ed)
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action during the developmental stages of an organism.
Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
A genus of BACILLACEAE that are spore-forming, rod-shaped cells. Most species are saprophytic soil forms with only a few species being pathogenic.
Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.
The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat.
Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein.
The commonest and widest ranging species of the clawed "frog" (Xenopus) in Africa. This species is used extensively in research. There is now a significant population in California derived from escaped laboratory animals.
The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Female germ cells derived from OOGONIA and termed OOCYTES when they enter MEIOSIS. The primary oocytes begin meiosis but are arrested at the diplotene state until OVULATION at PUBERTY to give rise to haploid secondary oocytes or ova (OVUM).
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.
A technique with which an unknown region of a chromosome can be explored. It is generally used to isolate a locus of interest for which no probe is available but that is known to be linked to a gene which has been identified and cloned. A fragment containing a known gene is selected and used as a probe to identify other overlapping fragments which contain the same gene. The nucleotide sequences of these fragments can then be characterized. This process continues for the length of the chromosome.
A set of three nucleotides in a protein coding sequence that specifies individual amino acids or a termination signal (CODON, TERMINATOR). Most codons are universal, but some organisms do not produce the transfer RNAs (RNA, TRANSFER) complementary to all codons. These codons are referred to as unassigned codons (CODONS, NONSENSE).
DNA constructs that are composed of, at least, a REPLICATION ORIGIN, for successful replication, propagation to and maintenance as an extra chromosome in bacteria. In addition, they can carry large amounts (about 200 kilobases) of other sequence for a variety of bioengineering purposes.
Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.
A group of genetically identical cells all descended from a single common ancestral cell by mitosis in eukaryotes or by binary fission in prokaryotes. Clone cells also include populations of recombinant DNA molecules all carrying the same inserted sequence. (From King & Stansfield, Dictionary of Genetics, 4th ed)
A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in nature. Some species are pathogenic for humans, animals, and plants.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in plants.
A species of gram-positive bacteria that is a common soil and water saprophyte.
Proteins found in any species of fungus.
Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely.
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.-.
Amino acid sequences found in transported proteins that selectively guide the distribution of the proteins to specific cellular compartments.
The class of all enzymes catalyzing oxidoreduction reactions. The substrate that is oxidized is regarded as a hydrogen donor. The systematic name is based on donor:acceptor oxidoreductase. The recommended name will be dehydrogenase, wherever this is possible; as an alternative, reductase can be used. Oxidase is only used in cases where O2 is the acceptor. (Enzyme Nomenclature, 1992, p9)
The part of CENTRAL NERVOUS SYSTEM that is contained within the skull (CRANIUM). Arising from the NEURAL TUBE, the embryonic brain is comprised of three major parts including PROSENCEPHALON (the forebrain); MESENCEPHALON (the midbrain); and RHOMBENCEPHALON (the hindbrain). The developed brain consists of CEREBRUM; CEREBELLUM; and other structures in the BRAIN STEM.
An aquatic genus of the family, Pipidae, occurring in Africa and distinguished by having black horny claws on three inner hind toes.
Proteins found in any species of insect.
The functional hereditary units of VIRUSES.
CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells.
A plant genus of the family BRASSICACEAE that contains ARABIDOPSIS PROTEINS and MADS DOMAIN PROTEINS. The species A. thaliana is used for experiments in classical plant genetics as well as molecular genetic studies in plant physiology, biochemistry, and development.
The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).
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.-.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
Chromosomes in which fragments of exogenous DNA ranging in length up to several hundred kilobase pairs have been cloned into yeast through ligation to vector sequences. These artificial chromosomes are used extensively in molecular biology for the construction of comprehensive genomic libraries of higher organisms.
Deoxyribonucleic acid that makes up the genetic material of viruses.
Yeast-like ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES isolated from exuded tree sap.
Multicellular, eukaryotic life forms of kingdom Plantae (sensu lato), comprising the VIRIDIPLANTAE; RHODOPHYTA; and GLAUCOPHYTA; all of which acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations.
Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).
A phenotypically recognizable genetic trait which can be used to identify a genetic locus, a linkage group, or a recombination event.
The male gonad containing two functional parts: the SEMINIFEROUS TUBULES for the production and transport of male germ cells (SPERMATOGENESIS) and the interstitial compartment containing LEYDIG CELLS that produce ANDROGENS.
The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.
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
A subclass of enzymes which includes all dehydrogenases acting on primary and secondary alcohols as well as hemiacetals. They are further classified according to the acceptor which can be NAD+ or NADP+ (subclass 1.1.1), cytochrome (1.1.2), oxygen (1.1.3), quinone (1.1.5), or another acceptor (1.1.99).
Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.
Annual cereal grass of the family POACEAE and its edible starchy grain, rice, which is the staple food of roughly one-half of the world's population.
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.
Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.
Partial cDNA (DNA, COMPLEMENTARY) sequences that are unique to the cDNAs from which they were derived.
Genes which regulate or circumscribe the activity of other genes; specifically, genes which code for PROTEINS or RNAs which have GENE EXPRESSION REGULATION functions.
Glycoproteins found on the membrane or surface of cells.
Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.
Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.
Any cell, other than a ZYGOTE, that contains elements (such as NUCLEI and CYTOPLASM) from two or more different cells, usually produced by artificial CELL FUSION.

Novel regulation of the homeotic gene Scr associated with a crustacean leg-to-maxilliped appendage transformation. (1/69323)

Homeotic genes are known to be involved in patterning morphological structures along the antero-posterior axis of insects and vertebrates. Because of their important roles in development, changes in the function and expression patterns of homeotic genes may have played a major role in the evolution of different body plans. For example, it has been proposed that during the evolution of several crustacean lineages, changes in the expression patterns of the homeotic genes Ultrabithorax and abdominal-A have played a role in transformation of the anterior thoracic appendages into mouthparts termed maxillipeds. This homeotic-like transformation is recapitulated at the late stages of the direct embryonic development of the crustacean Porcellio scaber (Oniscidea, Isopoda). Interestingly, this morphological change is associated with apparent novelties both in the transcriptional and post-transcriptional regulation of the Porcellio scaber ortholog of the Drosophila homeotic gene, Sex combs reduced (Scr). Specifically, we find that Scr mRNA is present in the second maxillary segment and the first pair of thoracic legs (T1) in early embryos, whereas protein accumulates only in the second maxillae. In later stages, however, high levels of SCR appear in the T1 legs, which correlates temporally with the transformation of these appendages into maxillipeds. Our observations provide further insight into the process of the homeotic leg-to-maxilliped transformation in the evolution of crustaceans and suggest a novel regulatory mechanism for this process in this group of arthropods.  (+info)

Mrj encodes a DnaJ-related co-chaperone that is essential for murine placental development. (2/69323)

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

Requirement of a novel gene, Xin, in cardiac morphogenesis. (3/69323)

A novel gene, Xin, from chick (cXin) and mouse (mXin) embryonic hearts, may be required for cardiac morphogenesis and looping. Both cloned cDNAs have a single open reading frame, encoding proteins with 2,562 and 1,677 amino acids for cXin and mXin, respectively. The derived amino acid sequences share 46% similarity. The overall domain structures of the predicted cXin and mXin proteins, including proline-rich regions, 16 amino acid repeats, DNA-binding domains, SH3-binding motifs and nuclear localization signals, are highly conserved. Northern blot analyses detect a single message of 8.9 and 5.8 kilo base (kb) from both cardiac and skeletal muscle of chick and mouse, respectively. In situ hybridization reveals that the cXin gene is specifically expressed in cardiac progenitor cells of chick embryos as early as stage 8, prior to heart tube formation. cXin continues to be expressed in the myocardium of developing hearts. By stage 15, cXin expression is also detected in the myotomes of developing somites. Immunofluorescence microscopy reveals that the mXin protein is colocalized with N-cadherin and connexin-43 in the intercalated discs of adult mouse hearts. Incubation of stage 6 chick embryos with cXin antisense oligonucleotides results in abnormal cardiac morphogenesis and an alteration of cardiac looping. The myocardium of the affected hearts becomes thickened and tends to form multiple invaginations into the heart cavity. This abnormal cellular process may account in part for the abnormal looping. cXin expression can be induced by bone morphogenetic protein (BMP) in explants of anterior medial mesoendoderm from stage 6 chick embryos, a tissue that is normally non-cardiogenic. This induction occurs following the BMP-mediated induction of two cardiac-restricted transcription factors, Nkx2.5 and MEF2C. Furthermore, either MEF2C or Nkx2.5 can transactivate a luciferase reporter driven by the mXin promoter in mouse fibroblasts. These results suggest that Xin may participate in a BMP-Nkx2.5-MEF2C pathway to control cardiac morphogenesis and looping.  (+info)

Mechanisms of GDF-5 action during skeletal development. (4/69323)

Mutations in GDF-5, a member of the TGF-beta superfamily, result in the autosomal recessive syndromes brachypod (bp) in mice and Hunter-Thompson and Grebe-type chondrodysplasias in humans. These syndromes are all characterised by the shortening of the appendicular skeleton and loss or abnormal development of some joints. To investigate how GDF-5 controls skeletogenesis, we overexpressed GDF-5 during chick limb development using the retrovirus, RCASBP. This resulted in up to a 37.5% increase in length of the skeletal elements, which was predominantly due to an increase in the number of chondrocytes. By injecting virus at different stages of development, we show that GDF-5 can increase both the size of the early cartilage condensation and the later developing skeletal element. Using in vitro micromass cultures as a model system to study the early steps of chondrogenesis, we show that GDF-5 increases chondrogenesis in a dose-dependent manner. We did not detect changes in proliferation. However, cell suspension cultures showed that GDF-5 might act at these stages by increasing cell adhesion, a critical determinant of early chondrogenesis. In contrast, pulse labelling experiments of GDF-5-infected limbs showed that at later stages of skeletal development GDF-5 can increase proliferation of chondrocytes. Thus, here we show two mechanisms of how GDF-5 may control different stages of skeletogenesis. Finally, our data show that levels of GDF-5 expression/activity are important in controlling the size of skeletal elements and provides a possible explanation for the variation in the severity of skeletal defects resulting from mutations in GDF-5.  (+info)

Molecular cloning and epitope analysis of the peanut allergen Ara h 3. (5/69323)

Peanut allergy is a significant IgE-mediated health problem because of the increased prevalence, potential severity, and chronicity of the reaction. Following our characterization of the two peanut allergens Ara h 1 and Ara h 2, we have isolated a cDNA clone encoding a third peanut allergen, Ara h 3. The deduced amino acid sequence of Ara h 3 shows homology to 11S seed-storage proteins. The recombinant form of this protein was expressed in a bacterial system and was recognized by serum IgE from approximately 45% of our peanut-allergic patient population. Serum IgE from these patients and overlapping, synthetic peptides were used to map the linear, IgE-binding epitopes of Ara h 3. Four epitopes, between 10 and 15 amino acids in length, were found within the primary sequence, with no obvious sequence motif shared by the peptides. One epitope is recognized by all Ara h 3-allergic patients. Mutational analysis of the epitopes revealed that single amino acid changes within these peptides could lead to a reduction or loss of IgE binding. By determining which amino acids are critical for IgE binding, it might be possible to alter the Ara h 3 cDNA to encode a protein with a reduced IgE-binding capacity. These results will enable the design of improved diagnostic and therapeutic approaches for food-hypersensitivity reactions.  (+info)

TIF1gamma, a novel member of the transcriptional intermediary factor 1 family. (6/69323)

We report the cloning and characterization of a novel member of the Transcriptional Intermediary Factor 1 (TIF1) gene family, human TIF1gamma. Similar to TIF1alpha and TIF1beta, the structure of TIF1beta is characterized by multiple domains: RING finger, B boxes, Coiled coil, PHD/TTC, and bromodomain. Although structurally related to TIF1alpha and TIF1beta, TIF1gamma presents several functional differences. In contrast to TIF1alpha, but like TIF1beta, TIF1 does not interact with nuclear receptors in yeast two-hybrid or GST pull-down assays and does not interfere with retinoic acid response in transfected mammalian cells. Whereas TIF1alpha and TIF1beta were previously found to interact with the KRAB silencing domain of KOX1 and with the HP1alpha, MODI (HP1beta) and MOD2 (HP1gamma) heterochromatinic proteins, suggesting that they may participate in a complex involved in heterochromatin-induced gene repression, TIF1gamma does not interact with either the KRAB domain of KOX1 or the HP1 proteins. Nevertheless, TIF1gamma, like TIF1alpha and TIF1beta, exhibits a strong silencing activity when tethered to a promoter. Since deletion of a novel motif unique to the three TIF1 proteins, called TIF1 signature sequence (TSS), abrogates transcriptional repression by TIF1gamma, this motif likely participates in TIF1 dependent repression.  (+info)

Anopheles gambiae Ag-STAT, a new insect member of the STAT family, is activated in response to bacterial infection. (7/69323)

A new insect member of the STAT family of transcription factors (Ag-STAT) has been cloned from the human malaria vector Anopheles gambiae. The domain involved in DNA interaction and the SH2 domain are well conserved. Ag-STAT is most similar to Drosophila D-STAT and to vertebrate STATs 5 and 6, constituting a proposed ancient class A of the STAT family. The mRNA is expressed at all developmental stages, and the protein is present in hemocytes, pericardial cells, midgut, skeletal muscle and fat body cells. There is no evidence of transcriptional activation following bacterial challenge. However, bacterial challenge results in nuclear translocation of Ag-STAT protein in fat body cells and induction of DNA-binding activity that recognizes a STAT target site. In vitro treatment with pervanadate (vanadate and H2O2) translocates Ag-STAT to the nucleus in midgut epithelial cells. This is the first evidence of direct participation of the STAT pathway in immune responses in insects.  (+info)

In vivo expression of the nucleolar group I intron-encoded I-dirI homing endonuclease involves the removal of a spliceosomal intron. (8/69323)

The Didymium iridis DiSSU1 intron is located in the nuclear SSU rDNA and has an unusual twin-ribozyme organization. One of the ribozymes (DiGIR2) catalyses intron excision and exon ligation. The other ribozyme (DiGIR1), which along with the endonuclease-encoding I-DirI open reading frame (ORF) is inserted in DiGIR2, carries out hydrolysis at internal processing sites (IPS1 and IPS2) located at its 3' end. Examination of the in vivo expression of DiSSU1 shows that after excision, DiSSU1 is matured further into the I-DirI mRNA by internal DiGIR1-catalysed cleavage upstream of the ORF 5' end, as well as truncation and polyadenylation downstream of the ORF 3' end. A spliceosomal intron, the first to be reported within a group I intron and the rDNA, is removed before the I-DirI mRNA associates with the polysomes. Taken together, our results imply that DiSSU1 uses a unique combination of intron-supplied ribozyme activity and adaptation to the general RNA polymerase II pathway of mRNA expression to allow a protein to be produced from the RNA polymerase I-transcribed rDNA.  (+info)

Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word cloning refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine. In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest, then treated with enzymes in the test tube to generate smaller DNA fragments. Subsequently, these fragments are then combined with vector DNA to generate recombinant DNA molecules. The recombinant DNA is then introduced into a host ...
Wang, J, Xu, R & Liu, A. (2014) IRDL cloning: a one-tube, zero-background, easy-to-use, directional cloning method improves throughput in recombinant DNA preparation. PLoS ONE. 2014 Sep 23; 9(9):e107907. PM ID: ...
The genome information is offering opportunities to manipulate genes, polygenic characters and multiple traits in plants. Although a number of approaches have been developed to manipulate traits in plants, technical hurdles make the process difficult. Gene cloning vectors that facilitate the fusion, overexpression or down regulation of genes in plant cells are being used with various degree of success. In this study, we modified gateway MultiSite cloning vectors and developed a hybrid cloning strategy which combines advantages of both traditional cloning and gateway recombination cloning. We developed Gateway entry (pGATE) vectors containing attL sites flanking multiple cloning sites and plant expression vector (pKM12GW) with specific recombination sites carrying different plant and bacterial selection markers. We constructed a plant expression vector carrying a reporter gene (GUS), two Bt cry genes in a predetermined pattern by a single round of LR recombination reaction after restriction ...
For positive-selection cloning of PCR products through blunt-end ligation. Allows direct cloning of PCR products without purification.. GenHunters PCR-TRAP® Cloning System is the most efficient method for cloning PCR products by blunt-end ligation. This system uses a third generation cloning vector that features positive selection for cloning PCR products (see figures next page). Only recombinant plasmids confer antibiotic resistance, making PCR-TRAP® extremely efficient. There is no need for any post-PCR manipulation before cloning, since a significant fraction of PCR products do not contain the 3 overhanging A (Clark, 1988, Nucleic Acids Res. 16:9677 ...
TY - JOUR. T1 - Molecular cloning and primary structure of Man9‐mannosidase from human kidney. AU - BAUSE, Ernst. AU - BIEBERICH, Erhard. AU - ROLFS, Andreas. AU - VÖLKER, Christof. AU - SCHMIDT, Bernhard. PY - 1993/10. Y1 - 1993/10. N2 - Man9‐mannosidase, a processing enzyme found in the endoplasmic reticulum (ER), catalyses the removal of three distinct mannose residues from peptide‐bound Man9‐GlcNAc2 oligosaccharides producing a single Man6 isomer [Bause, E., Breuer, W., Schweden, J., Roesser, R. & Geyer, R. (1992) Eur. J. Biochem. 208, 451-457]. We have isolated four Man9‐mannosidase‐specific clones from a human kidney cDNA library and used these to construct a full‐length cDNA of 3250 base pairs. A single open reading frame of 1875 nucleotides encodes a protein of approximately 71 kDa, consistent with data from immunological studies. Analysis of the coding sequence predicts that Man9‐mannosidase is a type II transmembrane protein consisting of a short cytoplasmic ...
View Notes - PCR from BME 3406 at University of Florida. Cloning genes Cloning results in the purification of a single fragment of DNA from exceedingly complex DNA molecules. Need DNA corresponding
PCR Cloning Protocols, moment version, updates and expands Bruce Whites best-selling PCR Cloning Protocols (1997) with the latest methods for DNA cloning and mutagenesis. right here the researcher will locate with no trouble reproducible equipment for all of the significant points of PCR use, together with PCR optimization, desktop courses for PCR primer layout and research, and novel adaptations for cloning genes of distinct features or starting place, with emphasis on lengthy distance PCR and GC-rich template amplification. additionally integrated are either traditional and novel enzyme-free and restrict site-free methods to clone PCR items right into a diversity of vectors, in addition to state of the art protocols to facilitate DNA mutagenesis and recombination, and to clone the difficult uncharacterized DNA flanking a recognized DNA fragment. ...
Nucleic acids can be isolated from cells for the purposes of further analysis by breaking open the cells and enzymatically destroying all other major macromolecules. Fragmented or whole chromosomes can be separated on the basis of size by gel electrophoresis. Short stretches of DNA or RNA can be amplified by PCR. Southern and northern blotting can be used to detect the presence of specific short sequences in a DNA or RNA sample. The term cloning may refer to cloning small DNA fragments (molecular cloning), cloning cell populations (cellular cloning), or cloning entire organisms (reproductive cloning). Genetic testing is performed to identify disease-causing genes, and gene therapy is used to cure an inheritable disease.. Transgenic organisms possess DNA from a different species, usually generated by molecular cloning techniques. Vaccines, antibiotics, and hormones are examples of products obtained by recombinant DNA technology. Transgenic plants are usually created to improve characteristics of ...
The pUC18 plasmid and pUC19 plasmid enable successful cloning of larger DNA fragments than the M13 mp18 RF Phage Vector. Because these cloning vectors contain a multiple cloning site at the lacZ region, recombinant plamids can be verified via blue/white colony screening using agar plates containing IPTG and X-Gal. Expression of target DNA is enabled by the presence of a lac promoter in the cloning vectors.. ...
Use SimVector to draw plasmid maps, perform restriction analysis and mapping. The plasmid drawing software also simulates cloning experiments such as gateway cloning, ta cloning and restriction cloning
TY - JOUR. T1 - Molecular cloning and functional expression of rat leukotriene A4hydrolase using the polymerase chain reaction. AU - Makitala, Naomasa. AU - Funku, Colin D.. AU - Imaic, Enyu. AU - Hoover, Richard L.. AU - Badra, Kamal F.. PY - 1992. Y1 - 1992. N2 - We isolated a cDNA encoding rat leukotriene A4 (LTA4) hydrolase from mesangial cells by the polymerase chain reaction according to the human amino acid sequence. The deduced amino acid sequence shows that rat LTA, hydrolase is a 609 amino acid protein with an M, 69 kDa. Comparison of human LTA4 hydrolase revealed 93% homology, and include zinc-binding motifs of aminopeptidases. COS-7 cells transfected with the cDNA revealed substantial LTA4 hydrolase activity, and their activities were abolished by preincubation with captopril, representing the first reported cDNA expression of recombinant enzyme in mammalian cells. RNA blot analysis indicated that LTA4 hydrolase was expressed in glomerular endothelial, epithelial and mesangial ...
A bacterial cloning system for mapping and analysis of complex genomes has been developed. The BAC system (for bacterial artificial chromosome) is based on Escherichia coli and its single-copy plasmid F factor. It is capable of maintaining human genomic DNA fragments of greater than 300 kilobase pairs. Individual clones of human DNA appear to be maintained with a high degree of structural stability in the host, even after 100 generations of serial growth. Because of high cloning efficiency, easy manipulation of the cloned DNA, and stable maintenance of inserted DNA, the BAC system may facilitate construction of DNA libraries of complex genomes with fuller representation and subsequent rapid analysis of complex genomic structure.. ...
pEASY®-Blunt Zero Cloning Kit,Cloning Vectors,Cloning and Mutagenesis System,Products,Beijing TransGen Biotech Co.Ltd,OverviewContents& storageCitations & referencesRelated ImagesDownloadOverviewDescriptionpEASY ®-Blunt Zero Cloning Vector c
Puzzling Cloning Problem With a Specific Vector - posted in Molecular Cloning: Hello, over the last couple of months I have been attempting to generate artificial aggrobacterial vectors containing DNA inserts that correspond to the tripartite genome of the Cowpea Chlorotic Mosaic Virus. These vectors will be used to generate mutant virions in-Plantae. I am using a vector provided to our lab by a collborator. I must use this vector for all cloning experiments. Its a large vector ( abou...
CopyControl cDNA, Gene and PCR Cloning Kit w/ Chemically Competent E. coli cells from EPICENTRE Biotechnologies,The CopyControl PCR Cloning Kits are designed to speed up the PCR cloning process and to ensure that all PCR products, regardless of sequence or type of polymerase used, are efficiently cloned. All PCR products including those that are difficult to clone by other PCR cloning methods or that m,biological,biology supply,biology supplies,biology product
Cloning a foreign gene into E-coli - posted in Molecular Cloning: Dear Friends, I am facing the problem with cloning. I have insert with 2.6Kb to be cloned into 13Kb E-coli vector . Vector has cloned strong promoter which is derived from the source same as the insert coming from. According to the strategy the desired will be cloned in front of promoter using PCR product digested either with same enzyme at both the ends(NcoI) or two enzymes(NcoI and SmaI). After digestion,ligation and tra...
For every Herceptin, for every Enbrel, Humira and Rituxan, and for every successful biotherapeutic product in the market today, there is a long history of its product development. We all hope that our target protein product will make a difference in both the therapeutic market and the lives it will eventually affect as it reaches its ultimate goal of becoming a human therapeutic. The reality is, in order for it to be a feasible therapeutic, the protein must first be manufactured. And one of the very first steps in determining the manufacturing capability of the product is the crucial step of choosing the production cell line that will produce this protein.. There are many challenges in single cell CHO cloning procedures to obtain valuable stable CHO production cell lines. We often rely on efficient cloning strategies and cloning media to execute such a crucial task in the developmental process of a recombinant protein product. An important aspect of cloning strategies is the cloning media ...
1. A degenerate polymerase chain reaction (PCR) homology screening procedure was applied to rat brain cDNA in order to identify novel genes belonging to the amiloride-sensitive Na+ channel and degenerin (NaC/DEG) family of ion channels. A single gene was identified that encodes a protein related to …
The biosynthesis of steroid hormones is an integral component of insect growth, development and reproduction. Although there is an abundance of biochemical data implicating both microsomal and mitochondrial cytochrome P450s in steroid metabolism, molecular genetic information on mitochondrial P450s is almost entirely limited to vertebrate sequences. In the current study, a degenerate polymerase chain reaction (PCR) primer was targeted to the highly conserved region of P450 genes that encodes the heme-binding decapeptide. Using a 5 rapid amplification of cDNA ends (RACE) approach, seven novel cytochrome P450 genes were isolated from Drosophila acanthoptera, including one sequence (CYP12B1) with high regional homology to vertebrate mitochondrial P450s. Sequence analysis of the conceptual translation of the full length gene, obtained by 5RACE, revealed an amphipathic NH2-terminus rich in basic and hydrophilic amino acids, a characteristic feature of mitochondrial P450s that distinguishes them ...
PRODUCTS | Cloning & Gene Analysis | KOD -Plus- Mutagenesis Kit | The category leader, continuing to create new value that contributes to society in the environment, healthcare, and high-function product fields.
During the last few decades, techniques for manipulating eukaryotic as well as prokaryotic DNA have witnessed a remarkable development. There are three main phases of the development of these techniques, which include recombinant DNA technology and gene cloning; polymerase chain reaction and DNA chips and microarrays. Using recombinant DNA technology, we can isolate and clone single copy of a gene or a DNA segment into an indefinite number of copies, all identical. This became possible, because bacteria, phages and plasmids reproduce in their usual style, even after insertion of foreign DNA, so that the inserted DNA also replicates faithfully with the parent DNA. This technique is called gene cloning. It involves the production of a large number of identical DNA molecules from a single ancestral DNA molecule. The essential characteristic of gene cloning is that the desired gene or DNA fragments must be selectively amplified resulting in a large increase in copy number of selected DNA sequences. ...
Gentaur molecular products has all kinds of products like :search , GenTarget \ pEco-T7-nHis PCR cloning kit: PCR cloning kit with a built-in vector (T7 promoter based) in provided cloning cells for E Coli expression of N-term His-tagged protein. \ IC-1001 for more molecular products just contact us
Semi-solid cloning of mammalian cells in 96-well plates using ClonaCell™ FLEX is an efficient cloning method that enables selective expansion of only those clones producing the protein of interest. Whether cloning CHO cells, hybridomas or other cell
Reverse genetics is used in many laboratories around the world and enables the creation of tailor-made influenza viruses with a desired genotype or phenotype. However, the process is not flawless, and difficulties remain during cloning of influenza gene segments into reverse genetics vectors (pHW2000, pHH21, pCAGGS). Reverse genetics begins with making cDNA copies of influenza gene segments and cloning them into bi-directional (pHW2000) or uni-directional plasmids (pHH21, pCAGGS) followed by transfection of the recombinant plasmid(s) to HEK-293 T or any other suitable cells which are permissive to transfection. However, the presence of internal restriction sites in the gene segments of many field isolates of avian influenza viruses makes the cloning process difficult, if employing conventional methods. Further, the genetic instability of influenza gene-containing plasmids in bacteria (especially Polymerase Basic 2 and Polymerase Basic 1 genes; PB2 and PB1) also leads to erroneous incorporation of
Molecular cloning and deduced amino acid sequences of the alpha- and beta- subunits of mammalian NAD(+)-isocitrate dehydrogenase.: A 153 bp fragment of the cDNA
The procedure of PCR cloning includes DNA preparation, digestion, ligation and transformation. This video shows the first part- insert PCR. Compared to other Taq-generated PCR inserts, Pfu DNA polymerase-generated PCR fragments will have fewer errors and is more commonly used for molecular cloning. - PCR Cloning (I) - Inserted Fragment Preparation - AbVideo™ - Support - Abnova
P-57. Generating and Sequencing Full-length cDNAs of Novel Human Genes Within the German cDNA Consortium. Ruth E. Wellenreuther, Stefan Wiemann, Daniela Heiss, Nina Claudino, Annemarie Poustka, Department of Molecular Genome Analysis, German Cancer Research Center, Im Neuenheimer Heidelberg, GERMANY. We generate human cDNA libraries that are enriched in full length clones i.e. from the translation start to the poly A tail. These libraries are used for a) systematic sequencing within the cDNA consortium of the Genome Project aiming at the identification and analysis of as many new genes as possible and b) for screening to isolate full length clones of partial genes. Libraries are created by directional cloning of cDNAs into plasmid vectors. Full-length enrichment is achieved via Clontechs SMART technology. This method is PCR-based, and in our modified strategy, we amplify and clone selective size windows of the cDNA fraction above 3 kb.Clones from the libraries generated within this project are ...
We evaluated the cloning efficiency of different size PCR products into three T-vector cloning systems:. the pGEM-T and pGEM-T Easy Vector Systems and the TOPO TA Cloning System.
Learn about Molecular Cloning. Topics cover isolation of gene fragment, screening, polymerase chain reaction, cloning vectors, and the isolation of plasmids.
Get fast and efficient molecular cloning with SBI-quickly and easily make a range of constructs or let SBI take care of making them for you
The complete nucleotide and amino acid sequence of GP-2 was determined initially by molecular cloning technique from peptide sequence for rat, dog and humans (12, 13, 18, 43). The cDNA encodes a protein of 530 aa in humans with 67% identity to rat and 72% to dog GP-2. The primary sequence encodes 10 (human) or 8 (dog) Asn-linked glycosylation sites. This is consistent with data for in vitro translation by reticulocyte lysate that yields a protein of 55 kDa and treatment with N-glycanase which reduces the molecular mass of canine GP-2 from 75 kDa to 52 kDa (12, 18). Rat GP-2 has been shown to possess 5 or 6 N-linked high mannose carbohydrate chains (16). The primary protein sequence is also post-translationally glycosylated on its carboxyl terminal to a GPI moiety in the membrane during passage through the Golgi to reach the ZG (11, 13, 17). This GPI moiety contains a phosphoethanolamine linker attached to the protein, a 4-5 sugar core and a phosphatidylinositol whose hydrocarbon chains insert ...
Andre SzyszkowskiPsy 13004/28/05 Cloning: Choice is Ethical Thousands of people a year are placed on the organ donors list. Thousands of people a year are diagnosed with diseases that are dubbed fatal unless a transplant or transfusion is given. This has created a large demand for some alternative method to the present donor practice. Research in the taboos cience of cloning seems to provide a viable method in which to aid the problem aforementioned and many others as well. But is it ethical? Cloning technology is expected to aid the result in several medical breakthroughs. It is thought that there may one day be a cure for cancer. This is because the cloning process helps us understand the process of cell differentiation. Theories exist that if a cure for cancer can be found, then further testing may lead to a cure for heart attacks and cloning organs for organ transplantation. Scientists believe that they may be able to treat heart attack victims by cloning their healthy heart cells and ...
44.38935 -68.20742 BAR HARBOR, Maine - New scientific evidence has heightened concerns about the safety of cloning human beings, an international authority warned yesterday.. We now believe that the potential to make a normal human baby with cloning is almost zero, said Dr. Rudolf Jaenisch, professor of biology at the Massachusetts Institute of Technology, at a genetics conference at Maines Jackson Laboratory.. Cloning a normal human is an illusion. There are major problems, and the problems are very serious.. Dr. Jaenisch, who helped pioneer gene transfer and cloning, said the public continues to be enthralled by stories of maverick doctors who supposedly plan to clone human beings.. Italian doctors in Rome, for instance, claim that they will clone a baby by next spring, with $500,000 provided by a couple whose infant died after heart surgery. An American firm called Clonaid has made repeated claims about a imminent human cloning.. Cloning a normal baby is utter nonsense, said Dr. ...
Introduction. Does cloning benefit or endanger society? Marley Gibbons-Balfour Case Study: Biology Contents Introduction 1 Background Science 2 Arguments against 4 Arguments for 7 Summary 10 Conclusion 11 Bibliography 12 Introduction Cloning has quickly become one of the most contentious issues in modern society, along with other issues like abortion, homosexuality and euthanasia. Due to the conflicted teachings and ideologies of many people in the world, there is no general consensus about cloning. Some people feel that it could benefit humans (through cures, through solving infertility and through knowledge), while others feel it could endanger humans and is a bad thing (due to ethical issues and due to being unaware of what could happen if it didnt work). Because of this, I have decided to investigate whether cloning actually does benefit or endanger society. I will go about this by collecting 6 sources (3 against cloning and 3 for cloning) and evaluating the evidence they present with the ...
The cloning of a novel murine gene that encodes a protein with lymphopoietic activity is described. The activity was initially identified in a coculture system of an adherent thymic stromal cell line with a medullary phenotype, Z210R.1, and day 15 fetal liver (6). Interestingly, the nonadherent lymphoid-like cells that grew in these cocultures phenotypically resembled B cells. A clonal lymphoid line (NAG8/7) was derived from long term cocultures, and this line expresses B220, 6C3 (BP-1), and is surface μ+, but κ chain negative. Importantly, NAG8/7 cells proliferated in response to conditioned medium from the Z210R.1 cell line and thus became the basis of a bioassay that allowed for the preliminary characterization of the growth factor and the eventual cloning of the cDNA encoding this activity.. The cDNA encoding the NAG8/7 growth activity was cloned from a library made from the Z210R.1 cell line. Based on the nucleotide sequence, the mature TSLP protein consists of 121 amino acids with 3 ...
Gateway compatible bacterial expression vector with T7 promoter, N- and C-terminal His tags and thrombin cleavage site; contains ccdB death cassette that is removed after recombinational cloning; kanamycin resistance in bacteria; recombinational cloning ...
The comprehensive study of protein structure and function, or proteomics, depends on the obtainability of full‐length cDNAs in species‐specific expression vectors and subsequent functional analysis of the expressed protein
In-Fusion EcoDry PCR Cloning Kits allow you to clone any PCR fragment into any linearized vector in a single step without restriction digestion of the PCR fragment, ligation or blunt-end polishing.
A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites - a standard feature of engineered plasmids. Restriction sites within an MCS are typically unique, occurring only once within a given plasmid. MCSs are commonly used during procedures involving molecular cloning or subcloning. Extremely useful in biotechnology, bioengineering, and molecular genetics, MCSs let a molecular biologist insert a piece of DNA or several pieces of DNA into the region of the MCS. This can be used to create transgenic organisms, also known as genetically modified organisms (GMOs). One bacterial plasmid used in genetic engineering as a plasmid cloning vector is pUC18. Its polylinker region is composed of several restriction enzyme recognition sites, that have been engineered into a single cluster (the polylinker). It has restriction sites for various restriction enzymes, including EcoRI, BamHI and PstI. Another vector used in genetic ...
TY - JOUR. T1 - Construction of a set of Saccharomyces cerevisiae vectors designed for recombinational cloning. T2 - Author correction. AU - Van Mullem, Vincent. AU - Wery, Maxime. AU - De Bolle, Xavier. AU - Vandenhaute, Jean. PY - 2004/1/30. Y1 - 2004/1/30. UR - U2 - 10.1002/yea.1064. DO - 10.1002/yea.1064. M3 - Comment/debate. AN - SCOPUS:1242273822. VL - 21. JO - Yeast. JF - Yeast. SN - 0749-503X. IS - 2. ER - ...
Both the colony formation rate (number of colonies) and the ratio of correct clones (cloning efficiency) in transformation are important determinants for efficient cloning of PCR fragments. Cell lysates from E. coli RecA− strains such as DH10B contain endogenous in vitro homologous recombination activity, and can be used to clone PCR fragments into vectors with homology regions. However, cloning with lysates from this strain is not efficient, particularly in the case of inserts with short homology lengths (approximately 15-20 bp), because of a lower colony formation rate [14]. An E. coli PPY strain that expresses an optimized λ prophage Red/ET recombination system circumvents this problem by increasing the colony formation rate during PCR fragment cloning [14]. To extend the utility of this method, I prepared SLiCE extracts from several E. coli laboratory strains with some modifications, and estimated the efficiency with which redox-related genes from Arabidopsis could then be cloned into ...
A method is provided for isolating and identifying a recombinant clone having a DNA segment therein coding for at least one desired heterologous polypeptide, at least a short amino acid sequence of which is known, by effecting cDNA synthesis on a mixture of mRNAs containing the mRNA coding for the desired polypeptide, isolating the resultant cDNA mixture, inserting the resultant cDNA into recombinant cloning vehicles, transforming hosts with the vehicles, separating the transformants and isolating and identifying a recombinant clone containing a DNA segment which is homologous over at least a portion thereof to at least one oligonucleotide probe specific for the DNA segment; wherein the probe is an extension of the nucleotide sequence of an oligonucleotide primer having a nucleotide sequence complementary to a region of the target mRNA coding for a portion of the known amino acid sequence, and is complementary to a longer region of the target mRNA coding for a longer portion of the known amino acid
A method is provided for isolating and identifying a recombinant clone having a DNA segment therein coding for at least one desired heterologous polypeptide, at least a short amino acid sequence of which is known, by effecting cDNA synthesis on a mixture of mRNAs containing the mRNA coding for the desired polypeptide, isolating the resultant cDNA mixture, inserting the resultant cDNA into recombinant cloning vehicles, transforming hosts with the vehicles, separating the transformants and isolating and identifying a recombinant clone containing a DNA segment which is homologous over at least a portion thereof to at least one oligonucleotide probe specific for the DNA segment; wherein the probe is an extension of the nucleotide sequence of an oligonucleotide primer having a nucleotide sequence complementary to a region of the target mRNA coding for a portion of the known amino acid sequence, and is complementary to a longer region of the target mRNA coding for a longer portion of the known amino acid
There have been several successes of cloning especially the birth of Dolly the sheep in 1997, which lived for six years. Nuclear transfer involves the fusion of somatic cells and enucleated egg cells. Cloning of pig cells will be the focus of this paper. Scientists have identified pig clones as a potential hope for the future because of the possibility of xenotransplantation. Xenotransplantation has resulted from the merging of cloning with an additional biotechnology technique of genetic engineering. In addition, cloning has led to new prospects of livestock breeding and advances in medical procedures. This paper will discuss the procedure involved in the cloning of pig cells. Cloning is a multi-step process that scientists have endeavored to advance for a long time. The success story of Dolly the sheep served as a breakthrough for the cloning of mammalian cells after the success of other species (Cibell 2002, p. 32). Cloning has its basis on the understanding of the processes involved in ...
TY - JOUR. T1 - Molecular cloning of the cDNA for the catalytic subunit of human DNA polymerase δ. AU - Yang, Chun li. AU - Chang, Long sheng. AU - Zhang, Peng. AU - Hao, Huiling. AU - Zhu, Lingyun. AU - Lan Toomey, N.. AU - Lee, Marietta Y.W.t.. N1 - Funding Information: This work was supported by National Institutes of Health Grant GM31973 to MYWTL and in part by grants from the National Cancer Institute (CA 54323) and the Bremer and Milheim Foundation to LSC. An account of this work was presented at the Cold Spring Harbor Meeting on EukaryorJc DNA Replication in September 1991. We thank Drs A. Sugino, A. Morrison and G. Pignede for copies of their papers in press.. PY - 1992/2/25. Y1 - 1992/2/25. N2 - The cDNA of human DNA polymerase δ was cloned. The cDNA had a length of 3.5 kb and encoded a protein of 1107 amino acid residues with a calculated molecular mass of 124 kDa. Northern blot analysis showed that the cDNA hybridized to a mRNA of 3.4 kb. Monoclonal and polyclonal antibodies to the ...
A DNA fragment from Bacillus natto IFO3936 has been cloned which enhances the production of both extracellular alkaline and neutral proteases in Bacillus subtilis. The DNA sequence analysis around the gene responsible for the hyperproduction, prtR, revealed one open reading frame (comprising 60 amino acid residues) which was bounded by potential transcriptional and translational regulatory signals in its preceding and following regions. This open reading frame was not homologous to the published sequences of the structural genes of the two proteases. The calculated molecular weight (7,109) of the polypeptide predicted from the DNA sequence is much smaller than those of the two proteases, indicating that the gene product is distinct from those enzymes. In-frame fusion between the N-terminal region of the coding sequence and the lacZ gene of Escherichia coli demonstrated that the coding region was indeed translated in vivo. By deletion analysis it was suggested that prtR was the structural gene ...
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Using the pYES vector from Invitrogen we first deleted five forbidden restriction sites in the vector back bone via side directed mutagenesis. Furthermore the original multiple cloning site was replaced for a multiple cloning site compatible to the RFC 10/25 cloning standards. To allow easy extraction and purification of proteins for in vitro applications the new multiple cloning site allows to express proteins with a Strep-tag II. Exclusion of the galactose inducible promoter provided a powerful basis vector for the integration of user-defined promoters. This way the pTUM100 vector gives a valuable contribution to our and to further protein expression and promoter characterization experiments in the yeast Saccharomyces cerevisiae. Moreover we used the pTUM100 to integrate the three constitutive promoters Tef1, Tef2 and ADH which come all with different promoter intensities. ...
Misc.Comments : Deposited by: Ichiro N. Maruyama Restriction digests of the clone give the following sizes (kb): HindIII--24.1, 17.6; EcoRI--22.0, 20.0; BamHI--24.8, 16.9; XbaI--32.7,9.0; NotI--41.7. (ATCC staff) Vector useful for constructing cDNA libraries. Permits positive selection for inserts using the Spi- phenotype, and excision of phagemid by lox/cre site-specific recombination. [1] To prepare phagemid from lambdaMGU2, grow recombinants on a RecA- host expressing the Cre protein (E. coli 1046[pCRE1], ATCC 77368) and select for ampicillin resistance. The pMGU product is 4.185 kb. [1] The order of the major features in the cloning region of the lambda vector is: lambda J - SmaI - SalI - loxP - EcoRI - M13 ori - ampR - pMB1 ori - HindIII - 3gam/BamHI/5gam - XhoI - loxP - SalI - lambda N. [1] Inserts can be amplified using the following primers flanking the BamHI cloning site: upstream 5-AAGAGGCAGAACTGGCAG-3 and downstream 5-ATCGATGCATAGCGATTC-3. [1] Efficiency of phagemid recovery is ...
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By Diego , source: Jan 31st, 2011 Well, unfortunately the weekend is over and the work week has begun. Today begins the five worst days of the week- the work days. At least you have us here at Daily Infographic to provide you with fun and interesting infographics. Todays infographic topic is cloning!. As long as I can remember there has been talk of cloning Personally I think cloning would be a pretty cool thing. I think it would be pretty cool to watch yourself, and even to interact with yourself. Hollywood has, of course, made many movies involving cloning. However, their cloning is as simple as entering a machine and there instantly being a copy of you. Science tells us that this is not in any way plausible and that cloning requires much more effort. If only everything were as simple as Hollywood makes it out to be.. As many people know, we really havent done that much in regards to cloning. In fact, we have only cloned a few animals and there hasnt even been a large of variety of animals ...
Introduction. Abbreviations and Acronyms. 1. FROM THE GENE TO THE TRANSGENIC ANIMAL.. Genome composition.. Gene structure.. The number of genes in genomes.. The major techniques of genetic engineering.. The systematic description of genomes.. Classical genetic selection.. Experimental mutation in genomes.. 2. TECHNIQUES FOR CLONING AND TRANSGENESIS.. Cloning.. Gene therapy.. Techniques of animal transgenesis.. 3. APPLICATIONS OF CLONING AND TRANSGENESIS.. Applications of animal cloning.. Applications of animal transgenesis.. 4. LIMITS AND RISKS OF CLONING, GENE THERAPY AND TRANSGENESIS.. Limits and risks of cloning.. Limits and risks of gene therapy.. Limits and risks of transgenesis.. Conclusion and Perspectives.. References.. Index. ...
Gene targeting is a powerful method that can be used for examining the functions of genes. Traditionally, the construction of knockout (KO) vectors requires an amplification step to obtain two homologous, large fragments of genomic DNA. Restriction enzymes that cut at unique recognitions sites and numerous cloning steps are then carried out; this is often a time-consuming and frustrating process. We have developed a one-step cloning method for the insertion of two arms into a KO vector using exonuclease III. We modified an adeno-associated virus KO shuttle vector (pTK-LoxP-NEO-AAV) to yield pAAV-LIC, which contained two cassettes at the two multiple-cloning sites. The vector was digested with EcoRV to give two fragments. The two homologous arms, which had an overlap of 16 bases with the ends of the vector fragments, were amplified by polymerase chain reaction. After purification, the four fragments were mixed and treated with exonuclease III, then transformed into Escherichia coli to obtain the desired
TY - JOUR. T1 - Structure of a family of rat amylase genes. AU - MacDonald, Raymond J.. AU - Crerar, Michael M.. AU - Swain, William F.. AU - Pictet, Raymond L.. AU - Thomas, Gilles. AU - Rutter, William J.. PY - 1980/12/1. Y1 - 1980/12/1. N2 - The sequences of two cloned rat pancreatic amylase cDNAs comprising 95% of the mRNA sequence are reported. Analysis of cloned rat genomic DNA fragments using cloned cDNA probes indicates that the rat genome contains multiple closely related amylase genes in which the cDNA sequences are distributed within a region 9 kilobases in length and are interrupted by at least seven intervening sequences.. AB - The sequences of two cloned rat pancreatic amylase cDNAs comprising 95% of the mRNA sequence are reported. Analysis of cloned rat genomic DNA fragments using cloned cDNA probes indicates that the rat genome contains multiple closely related amylase genes in which the cDNA sequences are distributed within a region 9 kilobases in length and are interrupted by ...
Molecular cloning, expression, functional characterization, chromosomal localization, and gene structure of junctate, a novel integral calcium binding protein of sarco(endo)plasmic reticulum membrane. In brain, 2.0 kb mRNA was highly expressed. A Chinese case of X-linked acrogigantism and systemic review. Hong S, Kim TW, Choi I, Woo JM, Oh J, Park WJ, Kim DH, Cho C. Biochim Biophys Acta. Get the latest public health information from CDC: While chromosome 19 only is the 19th largest autosomal chromosome, it contains 1440 protein-coding genes, and thus has the second highest number of protein-coding genes of any human chromosome. In brain, 2.0 kb mRNA was highly expressed. , M E Gåfvels, L G Paavola, C O Boyd, P M Nolan, F Wittmaack, A Chawla, M A Lazar, M Bucan, B O Angelin, J F Strauss, 3rd, Cloning of a complementary deoxyribonucleic acid encoding the murine homolog of the very low density lipoprotein/apolipoprotein-E receptor: expression pattern and assignment of ...
Scientists who do experimental genetics employ artificial selection experiments that permit the survival of organisms with user-defined phenotypes. Artificial selection is widely used in the field of microbial genetics, especially molecular cloning.. DNA recombination has been used to create gene replacements, deletions, insertions, inversions. Gene cloning and gene/protein tagging is also common. For gene replacements or deletions, usually a cassette encoding a drug-resistance gene is made by PCR.. Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word cloning refers to the fact that the method involves the replication of a single DNA molecule starting from a single living cell to generate a large population of cells containing identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the ...
This comprehensive yet balanced work emphasizes the principles and rationale underlying recombinant DNA methodology while furnishing a general understanding of the experimental protocols-suggesting flexible approaches to resolving particular molecular necessities that are easily adaptable to readers specific applications. Features summary tables presenting at-a-glance information on practices of recombinant DNA methodologies! Recombinant DNA Principles and Methodologies discusses basic and advanced topics requisite to the employment of recombinant DNA technology, such as plasmid biology nucleic acid biochemistry restriction enzymes cloning strategies gel electrophoresis southern and northern blotting preparation of probes phage lambda biology cosmids and genome analysis cloned gene expression polymerase chain reaction conventional and automated DNA sequencing site-directed mutagenesis and more! Elucidating the material with over 2250 edifying references, equations, drawings, and photographs, ...
Using a strategy based upon nucleotide sequence homology and starting from the sequence of the rat histamine H2 receptor (Ruat et al., Biochem. Biophys. Res. Commun. 1991, 179, 1470-1478), we have cloned a rat cDNA encoding a functional serotonin receptor (5-HT6). Its coding sequence corresponds to a glycoprotein of 436 amino acids displaying significant homology with other cloned monoaminergic receptors, e.g., various serotonin receptors. Genomic analysis of its gene indicated the presence of at least one intron. The major transcript of the 5-HT6 receptor gene has a size of approximately 4.1 kb but another minor 3.2 kb transcript was also evidenced. The highest expression, detected by Northern blot analysis as well as by in situ hybridization occurs in various serotoninergic areas of rat or guinea pig brain such as striatum, olfactory tubercle, nucleus accumbens and hippocampus, but a faint expression is also detectable in rat stomach. When transiently expressed in transfected COS-7 cells the 5-HT6
Cloning vectors A cloning vector is a small piece of DNA into which a foreign DNA fragment can be inserted. The insertion of the fragment into the cloning vector is carried out by treating...
Polymerase chain reaction and rapid amplification of cDNA ends were used to isolate cDNAs encoding a 5-hydroxytryptamine3 (5-HT3) receptor subunit and its splice variants from guinea pig intestine. The amino acid sequence predicted from this cDNA is 81% homologous to the murine 5-HT3 receptor subunits cloned from NCB20 and N1E-115 cells. The splice variants code for two proteins differing by a deletion of six amino acids located in the large intracellular loop between transmembrane domains M3 and M4. For characterization, the cloned 5-HT3 cDNA was expressed in HEK 293 cells, and the electrophysiological and pharmacological properties of the recombinant ion/channel/receptor complex were investigated by patch clamping. Our data reveal that the cloned cDNAs code for guinea pig 5-HT3 receptors, which functionally assemble as homo-oligomers. The kinetic behavior of the ion channel and its sensitivity to several agonists and antagonists were markedly different from those of the cloned 5-HT3 receptors from
Abstract 【Aim】 This study aims to clone a C-type lectin gene from Plutella xylostella, to investigate its expression patterns and to elucidate its agglutination on bacteria. 【Methods】 Based on the bioinformatical analysis of genome and transcriptome database of P. xylostella, the full-length cDNA of a C-type lectin gene was cloned from P. xylostella by RT-PCR and rapid amplification of cDNA ends (RACE) techniques. Prokaryotic expression plasmid was constructed and the fusion protein was expressed in E.coli BL21. The polyclonal antibody with high serum titer was prepared using the purified fusion protein to immunize New Zealand white rabbit. Real-time quantitative PCR (RT-qPCR) was employed to analyze the expression profiles of this gene in different tissues (hemocyte, cuticle, fat body, midgut and Malpighiam tubules) of the day-1 4th instar larvae and different developmental stages (egg, 1st-4th instar larva, prepupa, pupa, and adult) of P. xylostella. RT-qPCR and Western blot were ...
The majority of chromosomal translocations breakpoints are within regions of the genome where few DNA probes are available. The use of yeast artificial chromosomes (YACs) containing long stretches of human DNA allows dispersed DNA markers to be used to identify the position of breakpoints but does not readily allow subcloning of the precise breakpoint within the YAC DNA nor the cDNAs containing the affected genes. We describe a procedure allowing rapid isolation of cDNAs corresponding to genes within a YAC clone. Random cDNA is hybridised to PCR-generated biotinylated fragments of total DNA from a yeast strain harbouring a YAC clone. The hybrids can be recovered to facilitate subsequent cloning of the cDNA molecules. The application of this method to the cloning of cDNA molecules carrying sequences involved in the translocation t(4;11)(q21;q23) is illustrated.
With the completion of the human genome sequence the functional analysis and characterization of the encoded proteins has become the next urging challenge in the post-genome era. The lack of comprehensive ORFeome resources has thus far hampered systematic applications by protein gain-of-function analysis. Gene and ORF coverage with full-length ORF clones thus needs to be extended. In combination with a unique and versatile cloning system, these will provide the tools for genome-wide systematic functional analyses, to achieve a deeper insight into complex biological processes. Here we describe the generation of a full-ORF clone resource of human genes applying the Gateway cloning technology (Invitrogen). A pipeline for efficient cloning and sequencing was developed and a sample tracking database was implemented to streamline the clone production process targeting more than 2,200 different ORFs. In addition, a robust cloning strategy was established, permitting the simultaneous generation of two clone
pUC18 is a commonly used plasmid cloning vector in E.coli. Due to a small size pUC18 enables successful cloning of large DNA fragments. Ampicillin resistant. High copy number. Blue/White colony screening.
TY - JOUR. T1 - Molecular cloning and characterization of oocyte-specific Pat1a in Rana rugosa frogs. AU - Nakamura, Yoriko. AU - Iwasaki, Takehiro. AU - Umei, Yosuke. AU - Saotome, Kazuhiro. AU - Nakajima, Yukiko. AU - Kitahara, Shoichi. AU - Uno, Yoshinobu. AU - Matsuda, Yoichi. AU - Oike, Akira. AU - Kodama, Maho. AU - Nakamura, Masahisa. PY - 2015/10/1. Y1 - 2015/10/1. N2 - The Pat1 gene is expressed in the immature oocytes of Xenopus, and is reportedly involved in regulating the translation of maternal mRNAs required for oocyte-maturation. However, it is still unknown when Pat1a first appears in the differentiating ovary of amphibians. To address this issue, we isolated the full-length Pat1a cDNA from the frog Rana rugosa and examined its expression in the differentiating ovary of this frog. Among eight different tissues examined, the Pat1a mRNA was detectable in only the ovary. When frozen sections from the ovaries of tadpoles at various stages of development were immunostained for Vasa-a ...
ID PMB preliminary; circular DNA; SYN; 5590 BP. XX AC ATCC77385; XX DT 03-FEB-1994 (Rel. 8, Created) DT 01-JUL-1995 (Rel. 12, Last updated, Version 1) XX DE Vertebrate/E.coli plasmid vector pMB - incomplete. XX KW cloning vector. XX OS Cloning vector OC Artificial sequences; Cloning vehicles. XX RN [1] RC pS1 from pUC19 & oligo RC pS1-polyA from pS1 & pMAMneo RC pS1-MT from pS1 & pT24 RC pS1-MMTV from pS1 & pMAMneo RC pS3 from pS1 RC pS5 from pS3 RC pK from pUC19 & oligo RC pM from pK & pS1-polyA & pS1-MMTV, RSV LTR/MMTV LTR RC pT from pK & pS1-polyA & pS1-MT, MT gene RC pM-hyg from pM & pHT, hyg gene RC pMB-hyg from pM-hyg & pS5, lacZ-amb RC pMB from pMB-hyg RC pT-hyg from pT & pHT, hyg gene RC pTB-hyg from pT-hyg & pS5, lacZ-amb RC pTB from pTB-hyg RA Wang Q., Maher V.M., McCormick J.J.; RT Mammalian expression vectors with modulatable promoters and two RT multiple cloning sites; RL Gene 119:155-161(1992). XX CC Deposited by: J. Justin McCormick CC The first site was designed for insertion ...
February 6, 2003. SOUTH HADLEY, Mass.-Bacteria do it. Yeasts do it. Even some snails, shrimp, and aphids do it. But wait, while all of these creatures reproduce asexually through cloning, creating an exact replica of themselves, the cloning of more complex species, such as humans, still seems unnatural to many of us. Is it simply a case of getting used to a new technology, the way most of us got used to the idea of test tube babies over the past two decades? Or will reproductive cloning of humans ultimately be deemed unethical?. Questions such as these about cloning and stem-cell research-both for disease treatment and prevention and for reproduction-will be the focus of a series of events this spring on the theme, The Political Embryo: Reconceiving Human Reproduction, presented by Mount Holyoke Colleges Harriet L. and Paul M. Weissman Center for Leadership. In addition to a wide-ranging discussion of cloning, the series will look at the ethical and legal issues surrounding new and developing ...
Introduction. 3845 ??? Cloning, is it man playing god? Cloning has become currently one of the hottest topics in society because of the laboratory-produced sheep Dolly, named after the country singer Dolly Patron. The birth of this little cloned sheep shocked society tremendously and created many controversial discussions. In the wake of this human cloning, religious leaders all over the world have condemned the action citing the reason that it is playing of god. However, we must understand that we at the Consortium that are involved in this effort are as human as anyone else. There is almost no difference between in-vitro fertilization (IVF) and cloning, other than the fact that the source of the genetic material is slightly different. Cloning is not playing of god, rather it is a salutary act which can fulfill human needs. more. Middle. Looking back in science in the past, this distrust is totally understandable and logical. As an example, people think about atomic energy and above all ...
The risks of animal cloning are immense. The cloning process is inefficient and cloned animals have been observed to have higher rates of infection, tumour growth, and skeletal abnormalities than normal offspring. Are the risks and disadvantages of cloning because it is a nascent technology that scientists are trying to get to grips with, or are there inherent problems with the cloning process?
View Notes - unit7 cloning animals from BIOCHEM 100 at UMass (Amherst). UNIT 7 Animal Cloning and Epigenetics F08 The first cloned horse. What cell parts would you need to clone a human? (or
Obtaining the molecular clone of a gene can lead to the development of organisms that produce the protein product of the cloned ... In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest, then treated ... Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and ... In standard molecular cloning experiments, the cloning of any DNA fragment essentially involves seven steps: (1) Choice of host ...
Clark DP (2005). Molecular Biology. Academic Press. p. 611. ISBN 0-12-175551-7. "Addgene: What is a Plasmid?". ... This method can also be used to add new restriction sites to a multiple cloning site. Multiple cloning sites are a feature that ... A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction ... An MCS is found in a variety of vectors, including cloning vectors to increase the number of copies of target DNA, and in ...
... is a molecular biology technique in which DNA fragments are cloned into specific vectors without the requirement ... "Zero Blunt® Cloning Kits". (Cloning, Molecular biology). ... For "blunt end" TOPO cloning, the recipient vector does not have overhangs and blunt-ended DNA fragments can be cloned. The ... Commercial kits, such as the Zero Blunt® Cloning Kit from Invitrogen, are available. "The technology behind TOPO® Cloning". ...
... cloning is the process of creating cloned organisms (copies) of cells and of DNA fragments (molecular cloning). Coined by ... Two commonly discussed types of theoretical human cloning are therapeutic cloning and reproductive cloning. Therapeutic cloning ... Molecular cloning refers to the process of making multiple molecules. Cloning is commonly used to amplify DNA fragments ... "Cloning". Internet Encyclopedia of Philosophy. Cloning Fact Sheet from Human Genome Project Information website. 'Cloning' ...
Vector (molecular biology) Plant transformation vector IMAGE cDNA clones fosmid Golden Gate Cloning "Definition of cloning ... for example as used in the Gateway cloning system. The gene, once cloned into the cloning vector (called entry clone in this ... Cloning vectors in yeast include yeast artificial chromosomes (YACs). All commonly used cloning vectors in molecular biology ... Cloning vectors without promoter and RBS for the cloned DNA sequence are sometimes used, for example when cloning genes whose ...
"Expression cloning". Methods in Molecular Medicine. 94: 91-106. doi:10.1385/1-59259-679-7:91. ISBN 1-59259-679-7. PMID 14959824 ... Expression cloning is a technique in DNA cloning that uses expression vectors to generate a library of clones, with each clone ... Molecular cell biology genetics gene expression Transcription (genetics) translation λ phage pBR322 Lodes, M. J.; Dillon, D. C ... Usually the ultimate aim of expression cloning is to produce large quantities of specific proteins. To this end, a bacterial ...
Molecular and General Genetics. 204 (1): 108-114. doi:10.1007/bf00330196. PMID 3091993. S2CID 11992591 Gilley, David; Blackburn ... Cloning List of animals that have been cloned Polyclonal antibodies Polyclonal response Tumour heterogeneity "Clone definition ... This concept of clone assumes importance as all the cells that form a clone share common ancestry, which has a very significant ... Thus there are terms like polyclonal-derived from many clones; oligoclonal-derived from a few clones; and monoclonal-derived ...
... is a molecular cloning technique that relies on prior knowledge of the encoded protein's sequence or ... Positional cloning is another molecular cloning technique for identification of a gene of interest. This method uses exact ... This is not a requirement with functional cloning. TOPO Cloning is a cloning method that uses Taq polymerase. This is because ... Gateway recombination cloning is a cloning method in which a DNA fragment is moved from one plasmid backbone to another via a ...
2012). Tietz Textbook of Clinical Biochemistry and Molecular diagnostics. USA: Elsevier Saunders. p. 393. ISBN 9781416061649. ( ... A cloned enzyme donor immunoassay (CEDIA) is a competitive homogenous enzyme immunoassay. This assay makes use of two component ...
TOPO cloning (Cloning, Molecular biology, Biotechnology, Molecular biology techniques). ... TA cloning (also known as rapid cloning or T cloning) is a subcloning technique that avoids the use of restriction enzymes and ... The major downside of TA cloning is that directional cloning is not possible, so the gene has a 50% chance of getting cloned in ... "Improved TA Cloning". Caister Academic Press. Archived from the original on 2011-07-08. Retrieved 2009-10-17. "TA Cloning". www ...
complementary DNA (cDNA) cDNA library expressed sequence tags (EST) vector (molecular biology) cloning vector "I.M.A.G.E. ... IMAGE cDNA clones are a collection of DNA vectors containing cDNAs from various organisms including human, mouse, rat, non- ... IMAGE stands for integrated molecular analysis of genomes and their expression. From 1993 to 2007, the cDNA library was ...
... (LIC) is a form of molecular cloning that is able to be performed without the use of restriction ... "Get Your Clone 90% Of The Time with Ligation Independent Cloning". Bitesize Bio. Retrieved 2012-05-09. Haun, RS; Serventi, IM; ... LIC Primer Design (Wikipedia articles needing clarification from October 2011, Cloning, Molecular biology). ... "Seamless Cloning , NEB". Retrieved 2020-01-23. "Ligation Independent Cloning (LIC)". New England BioLabs (NEB). ...
Sambrook, Joseph; Russell, David W. (2001). "Commonly Used Techniques in Molecular Cloning". Molecular Cloning. 3. Li, Richard ... v t e (Molecular biology, Biochemistry methods, All stub articles, Molecular biology stubs). ... Phenol-chloroform extraction is a liquid-liquid extraction technique in molecular biology used to separate nucleic acids from ...
Molecular cloning is used to isolate and then transfer a DNA sequence of interest into a plasmid vector. This recombinant DNA ... Molecular biology is the study of the molecular underpinnings of the biological phenomena, focusing on molecular synthesis, ... "Foundations of Molecular Cloning - Past, Present and Future , NEB". Retrieved 2021-11-04. Alberts B, Johnson A, ... "Foundations of Molecular Cloning - Past, Present and Future , NEB". Retrieved 2021-11-25. " ...
Molecular Cloning. (4th ed. ed.). Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Pr. ISBN 1936113422. Forensic Biology ... Pääbo S (March 1989). "Ancient DNA: extraction, characterization, molecular cloning, and enzymatic amplification". Proceedings ... The Hirt extraction process gets rid of the high molecular weight nuclear DNA, leaving only low molecular weight mitochondrial ... It is a molecular method used, among other things, to recognize and count particular bacterial groupings. To recognize, define ...
Ohara, O. (2009). "ORFeome Cloning". Reverse Chemical Genetics. Methods in Molecular Biology. Vol. 577. pp. 3-9. doi:10.1007/ ... In, molecular genetics, an ORFeome refers to the complete set of open reading frames (ORFs) in a genome. The term may also be ... Complete ORF sets have been cloned for a number of organisms including Brucella melitensis,Chlamydia pneumoniae,Escherichia ... a resource for comparative molecular microbiology". BMC Genomics. 11: 470. doi:10.1186/1471-2164-11-470. PMC 3091666. PMID ...
Articles with short description, Short description matches Wikidata, Cloning, Molecular biology, Cryobiology, Applied genetics) ... Partial cloning also avoids the ethical problems associated with "classical" cloning in that it does not result in live born - ... "Partial cloning" (given by the red arrow) rejuvenates old cells without passage through an embryonic stage."Partial cloning" ( ... The measure of Diagram showing the difference between "Classical" and "Partial" cloning: Classical cloning (the route given by ...
Cloning, Molecular biology). ... It can be used as a type of cloning vector in combination with ... Similarly to a plasmid, a phagemid can be used to clone DNA fragments and be introduced into a bacterial host by a range of ... A phagemid or phasmid is a DNA-based cloning vector, which has both bacteriophage and plasmid properties. These vectors carry, ... Phagemids are used in a variety of biotechnology applications; for example, they can be used in a molecular biology technique ...
Molecular cloning and characterization". J Biol Chem. 277 (45): 43474-80. doi:10.1074/jbc.M206065200. PMID 12213818. "Entrez ...
... acts as a cytokine and its molecular structure is identical in humans, mice and rats. The bone morphogenetic protein ... Cloning and Stem Cells. 11 (3): 427-435. doi:10.1089/clo.2009.0024. PMID 19751112. Gamer LW, Wolfman NM, Celeste AJ, Hattersley ... Molecular Therapy. 24 (11): 1926-1938. doi:10.1038/mt.2016.160. PMC 5154476. PMID 27502608. Dai Z, Song G, Balakrishnan A, Yang ... International Journal of Molecular Sciences. 21 (7): 2598. doi:10.3390/ijms21072598. PMC 7177281. PMID 32283613. Ma, Y.; Liu, Y ...
Methods in Molecular Biology™. Methods in Molecular Biology. Vol. 23. Humana Press. pp. 9-22. doi:10.1385/0-89603-248-5:9. ISBN ... ISBN 978-0-87969-740-2. Messing J (1993). "M13 Cloning Vehicles" (PDF). In Griffin H.G., Griffin A.M. (eds.). DNA Sequencing ... Journal of Molecular Biology. 411 (5): 972-985. doi:10.1016/j.jmb.2011.07.002. ISSN 0022-2836. Smeal SW, Schmitt MA, Pereira RR ...
Messing, Joachim (1996). "Cloning Single-Stranded DNA". Molecular Biotechnology. 5 (1): 39-47. doi:10.1007/BF02762411. PMID ... Messing, Joachim (1991). "Cloning in M13 phage or how to use biology at its best". Gene. 100: 3-12. doi:10.1016/0378-1119(91) ... The simplicity of this family makes it an attractive model system to study fundamental aspects of molecular biology, and it has ... IKe and fd gene 5 proteins form left-handed helices with single-stranded DNA". Journal of Molecular Biology. 208 (1): 57-64. ...
Molecular cloning and expression". Eur. J. Biochem. 191 (3): 619-625. doi:10.1111/j.1432-1033.1990.tb19166.x. PMID 2390989. ... by isolation of genomic clones and complementary DNA clones utilizing polymerase chain reaction". J. Biol. Chem. 265 (2): 1102- ... 1992). "Molecular analysis of human glycophorin MiIX gene shows a silent segment transfer and untemplated mutation resulting ...
Kitagawa H, Uyama T, Sugahara K (Oct 2001). "Molecular cloning and expression of a human chondroitin synthase". J Biol Chem. ... Kitagawa H, Izumikawa T, Uyama T, Sugahara K (2003). "Molecular cloning of a chondroitin polymerizing factor that cooperates ... 2003). "Chondroitin sulfate synthase-3. Molecular cloning and characterization". J. Biol. Chem. 278 (41): 39711-25. doi:10.1074 ... The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro". DNA Res. 6 (1): 63-70. doi: ...
Molecular cloning and expression". Biochem. J. 329 ( Pt 2) (Pt 2): 275-82. doi:10.1042/bj3290275. PMC 1219041. PMID 9425109. ...
In molecular cloning, a vector is any particle (e.g., plasmids, cosmids, Lambda phages) used as a vehicle to artificially carry ... Lodish H, Berk A, Zipursky SL, Matsudaira P, Baltimore D, Darnell J (2000). "DNA Cloning with Plasmid Vectors". Molecular Cell ... Cloning site: This may be a multiple cloning site or other features that allow for the insertion of foreign DNA into the vector ... Some cloning vectors need not have a promoter for the cloned insert but it is an essential component of expression vectors so ...
European Molecular Biology Laboratory - Hamburg. Retrieved 2008-06-07. Russell DW, Sambrook J (2001). Molecular cloning: a ... These enzymes are routinely used for DNA modification in laboratories, and they are a vital tool in molecular cloning. The term ... Clark DP (2005). Molecular biology. Amsterdam: Elsevier Academic Press. ISBN 0-12-175551-7. Goodsell DS (2002). "The molecular ... this characteristic is widely used to perform in-vitro cloning techniques such as Golden Gate cloning. These enzymes may ...
Iwahana H, Oka J, Mizusawa N, Kudo E, Ii S, Yoshimoto K, Holmes EW, Itakura M (Jan 1993). "Molecular cloning of human ... Progress in Nucleic Acid Research and Molecular Biology. Vol. 42. pp. 259-87. doi:10.1016/s0079-6603(08)60578-4. ISBN ...
A plasmid is a double stranded circular DNA molecule commonly used for molecular cloning. Plasmids are generally 2 to 4 ... Once a clone from a genomic library is sequenced, the sequence can be used to screen the library for other clones containing ... ISBN 978-0-8053-9592-1. Russell, David W.; Sambrook, Joseph (2001). Molecular cloning: a laboratory manual. Cold Spring Harbor ... This is particularly determined by the number of clones needed to have in a library. The number of clones to get a sampling of ...
2012) "Labeling of DNA Probes by Nick Translation". Molecular Cloning: A Laboratory Manual. Park, K; Kim, J; Lim, S; Han, S; ... Next, the clones are sequenced and their position on the genome is verified. Probe labelling can be carried out by using either ... DNA is extracted from the BAC clones and amplified using a polymerase-based technique, such as degenerate oligonucleotide ... Shizuya, H; Kouros-Mehr, H (2001). "The development and applications of the bacterial artificial chromosome cloning system". ...
Suchi M, Mizuno H, Kawai Y, Tsuboi T, Sumi S, Okajima K, Hodgson ME, Ogawa H, Wada Y (Mar 1997). "Molecular cloning of the ... Suchi M, Harada N, Tsuboi T, Asai K, Okajima K, Wada Y, Takagi Y (1990). "Molecular cloning of human UMP synthase". Purine and ... Suttle DP, Bugg BY, Winkler JK, Kanalas JJ (Mar 1988). "Molecular cloning and nucleotide sequence for the complete coding ... Progress in Nucleic Acid Research and Molecular Biology. Vol. 53. pp. 1-78. doi:10.1016/s0079-6603(08)60142-7. ISBN ...
Multiple cloning sites are sometimes used to ensure that the fragments are inserted in all three possible reading frames so ... Initial work was done by laboratories at the MRC Laboratory of Molecular Biology (Greg Winter and John McCafferty), the Scripps ... This displayed the different peptides on the outer surfaces of the collection of viral clones, where the screening step of the ... Phage display technology was further developed and improved by groups at the Laboratory of Molecular Biology with Greg Winter ...
2016) Molecular ecolution of the capsid gene in human norovirus genogroup II. Sci Rep 6:29400 Ozaki K, Matsushima Y, Nagasawa K ... The cloning and sequencing of the Norwalk virus genome showed that these viruses have a genomic organization consistent with ... fragment-based molecular docking and binding free energy calculations". Carbohydr. Res. 378: 133-8. doi:10.1016/j.carres. ... Motoya T, Ryo A, Kuroda M, Katayama K, Kimura H (2018) Molecular evolutionary analyses of the RNA-dependent RNA polymerase ...
"Cloning and mapping of a cDNA for methionine synthase reductase, a flavoprotein defective in patients with homocystinuria". ... "Human methionine synthase reductase is a molecular chaperone for human methionine synthase". Proceedings of the National ...
Lee J, Wang Z, Luoh SM, Wood WI, Scadden DT (January 1994). "Cloning of FRK, a novel human intracellular SRC-like tyrosine ... Molecular and Cellular Biology. 10 (3): 1000-9. doi:10.1128/mcb.10.3.1000. PMC 360952. PMID 1689455. Zan L, Wu H, Jiang J, Zhao ... Nada S, Okada M, MacAuley A, Cooper JA, Nakagawa H (May 1991). "Cloning of a complementary DNA for a protein-tyrosine kinase ... Oberg-Welsh C, Welsh M (January 1995). "Cloning of BSK, a murine FRK homologue with a specific pattern of tissue distribution ...
1331-1342 Collins PL, Hill MG... Murphy BR (1995). Production of infectious human respiratory syncytial virus from cloned cDNA ... American molecular biologists, 20th-century American scientists, Vaccinologists, Wesleyan University alumni, National ...
Mukae N, Enari M, Sakahira H, Fukuda Y, Inazawa J, Toh H, Nagata S (August 1998). "Molecular cloning and characterization of ... May 2004). "Structural mechanism for inactivation and activation of CAD/DFF40 in the apoptotic pathway". Molecular Cell. 14 (4 ... 2007). "Large-scale mapping of human protein-protein interactions by mass spectrometry". Molecular Systems Biology. 3 (1): 89. ... Journal of Molecular Biology. 297 (5): 1121-8. doi:10.1006/jmbi.2000.3643. PMID 10764577. PDB: 1V0D​; Woo EJ, Kim YG, Kim MS, ...
Irie Y, Yamagata K, Gan Y, Miyamoto K, Do E, Kuo CH, Taira E, Miki N (2000). "Molecular cloning and characterization of Amida, ...
DNA was purified, amplified, and the PCR products cloned. The isolates used for molecular comparison are listed and consists of ... The molecular characterization was done by several molecular methods which include sequence-specific multiplex PCR, PCR-RFLP of ... Due to the lack of morphological measurements of the isolates from South America to go with the molecular results from the US ... Morphological and molecular characterization of Globodera populations from Oregon and Idaho. Phytopathology 101:480-491. Handoo ...
X's cloning machine to malfunction and free the Negative 4, who wants revenge for what the Super 4 did to them, so they kidnap ... the doctor's legs and rejoin the two pieces before they are damaged because the slightest bobo will modify the body's molecular ... Negatives Doctor X has made a cloning machine but it makes negative copies of the 4 who are as evil as the originals are noble ... Their evil doppelgangers set about making a clone army to take over the world. 43. All That Glitters Ruby seeks a hoard of ...
The luciferase enzyme consists of a 555-amino acid-long peptide with a molecular mass of 61627 u, while the luciferine vargulin ... 2004). "cDNA Cloning and Characterization of a Secreted Luciferase from the Luminous Japanese Ostracod, Cypridina noctiluca". ... "Cloning and expression of cDNA for the luciferase from the marine ostracod Vargula hilgendorfii". Proceedings of the National ... Molecular Biology and Evolution. 22 (7): 1543-1545. doi:10.1093/molbev/msi155. PMID 15858206. Shimomura, Osamu (2006). "The ...
"Molecular cloning and structural analysis of a gene from Zea mays (L.) coding for a putative receptor for the plant hormone ... In molecular biology, the auxin binding protein family is a family of proteins which bind auxin. They are located in the lumen ... Palme K, Hesse T, Campos N, Garbers C, Yanofsky MF, Schell J (February 1992). "Molecular analysis of an auxin binding protein ...
Molecular Medicine Reports. 25 (1). doi:10.3892/mmr.2021.12533. PMC 8628290. PMID 34791506. Xiao Y, Yuan Y, Zhang Y, Li J, Liu ... by in silico cloning and experimental validation". Genomics. 81 (6): 609-17. doi:10.1016/s0888-7543(03)00095-8. PMID 12782130. ...
Like the Ten-m family, Ten-m3 is a large type II transmembrane glycoprotein that has a molecular weight of ~300 kDa and is ... Qian X, Barsyte-Lovejoy D, Wang L, Chewpoy B, Gautam N, Al Chawaf A, Lovejoy DA (June 2004). "Cloning and characterization of ... Ten-m3 protein is expressed in the Purkinje's cell zone, molecular and granular layers and the white matter of the cerebellum. ... Molecular Brain Research. 133 (2): 253-65. doi:10.1016/j.molbrainres.2004.10.019. PMID 15710242. Ben-Zur, T.; Feige, E.; Motro ...
Arm JP, Nwankwo C, Austen KF (September 1997). "Molecular identification of a novel family of human Ig superfamily members that ... Samaridis J, Colonna M (March 1997). "Cloning of novel immunoglobulin superfamily receptors expressed on human myeloid and ... Norman PJ, Carey BS, Stephens HA, Vaughan RW (June 2003). "DNA sequence variation and molecular genotyping of natural killer ...
Cooper GM (2000). "The Molecular Composition of Cells". The Cell: A Molecular Approach (2nd ed.). Archived from the original on ... Lees ND, Skaggs B, Kirsch DR, Bard M (March 1995). "Cloning of the late genes in the ergosterol biosynthetic pathway of ... Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P (2002). "How Cells Obtain Energy from Food". Molecular Biology of ... From metabolites to molecular genetics". Diabetes Care. 13 (6): 582-99. doi:10.2337/diacare.13.6.582. PMID 2162755. S2CID ...
Imprinting may cause problems in cloning, with clones having DNA that is not methylated in the correct positions. It is ... Methods in Molecular Biology. Vol. 2093. New York, NY: Springer US. pp. 173-201. doi:10.1007/978-1-0716-0179-2_13. ISBN 978-1- ... If time is the responsible factor, it may be possible to delay cell division in clones, giving time for proper reprogramming to ... However, our understanding of the molecular mechanisms behind genomic imprinting show that it is the maternal genome that ...
Yabe T, McSherry C, Bach FH, Houchins JP (October 1990). "A cDNA clone expressed in natural killer and T cells that likely ... Hanson DA, Kaspar AA, Poulain FR, Krensky AM (May 1999). "Biosynthesis of granulysin, a novel cytolytic molecule". Molecular ... Harris V, Jackson C, Cooper A (December 2016). "Review of Toxic Epidermal Necrolysis". International Journal of Molecular ... The path to transcription has not been elucidated: transcription factors, promoter regions, and pathogen-associated molecular ...
Gridley T, Gray DA, Orr-Weaver T, Soriano P, Barton DE, Francke U, Jaenisch R (May 1990). "Molecular analysis of the Mov 34 ... Tsurumi C, DeMartino GN, Slaughter CA, Shimbara N, Tanaka K (May 1995). "cDNA cloning of p40, a regulatory subunit of the human ... The interaction of S4 with S5 and S12". Journal of Molecular Biology. 165 (2): 357-74. doi:10.1016/S0022-2836(83)80261-7. PMID ... Gödderz D, Dohmen RJ (Feb 2009). "Hsm3/S5b joins the ranks of 26S proteasome assembly chaperones". Molecular Cell. 33 (4): 415- ...
DNA cloning can also be performed intentionally by laboratory researchers. Here, DNA fragmentation is a molecular genetic ... In cell biology, ways in which fragmentation is useful for a cell: DNA cloning and apoptosis. DNA cloning is important in ... The key to cloning a DNA fragment is to link it to a vector DNA molecule that can replicate within a host cell. After a single ... Molecular Cell Biology. 7th ed. New York: W.H. Freeman and, 2013. Print. Bortner, Carl D., Nicklas B.E. Oldenburg, and John A. ...
1997). "Molecular cloning of cDNA encoding rat very long-chain acyl-CoA synthetase". J. Biol. Chem. 271 (48): 30360-5. doi: ... 1999). "Human very-long-chain acyl-CoA synthetase: cloning, topography, and relevance to branched-chain fatty acid metabolism ... cDNA cloning and characterization of a second enzymatically active protein". Mol. Genet. Metab. 68 (1): 32-42. doi:10.1006/mgme ...
Taniguchi T, Fujii-Kuriyama Y, Muramatsu M (July 1980). "Molecular cloning of human interferon cDNA". Proceedings of the ... Gene cloning also confirmed that IFN-α was encoded by a family of many related genes. The type II IFN (IFN-γ) gene was also ... to clone the human beta interferon gene in bacteria and the recombinant interferon was developed as 'betaseron' and approved ... By the early 1980s, genes for these interferons had been cloned, adding further definitive proof that interferons were ...
Fonagy A, Henning D, Jhiang S, Haidar M, Busch RK, Larson R, Valdez B, Busch H (1989). "Cloning of the cDNA and sequence of the ... "Identification and characterization of a human proliferation-associated nucleolar antigen with a molecular weight of 120,000 ... "Identification and characterization of a human proliferation-associated nucleolar antigen with a molecular weight of 120,000 ...
Chardin P, Madaule P, Tavitian A (March 1988). "Coding sequence of human rho cDNAs clone 6 and clone 9". Nucleic Acids Research ... Molecular and Cellular Biology. 20 (16): 6105-13. doi:10.1128/MCB.20.16.6105-6113.2000. PMC 86086. PMID 10913192. Michaelson D ... "Coding sequence of human rho cDNAs clone 6 and clone 9". Nucleic Acids Research. 16 (6): 2717. doi:10.1093/nar/16.6.2717. PMC ... X. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro". DNA Research. 5 (3): ...
2/2003, p. 425-453 LEWIS C. INGRAM, M. H. RICHMOND, AND R. B. SYKES: "Molecular Characterization of the R Factors Implicated in ... "Construction and use of a versatile set of broad-host-range cloning and expression vectors based on the RK2 replicon", Appl. ... Journal of Molecular Biology, Volume 239, Issue 5, 23 June 1994, Pages 623-663 Sequence data deposited at the NCBI: https://www ... "Molecular vehicle properties of the broad host range plasmid RK2", Science, December 1975: pp. 1226-1228. ...
Darwin, X-23, and Synch also learn that any Children who have died are resurrected by the City through cloning to further the ... Serafina is also able to detect genetic and bio-chemical data down to the molecular level. This makes her an extraordinary ... She can also manipulate bio-molecular ingredients to some degree. Aguja (Needle): She possesses powers that include the ...
Cloning from somatic cells rather than germ cells may begin life with a higher initial load of damage. Dolly the sheep died ... Evolutionary theories of aging primarily explain why aging happens, but do not concern themselves with the molecular mechanism( ... Molecular Phylogenetics and Evolution. 107: 232-238. doi:10.1016/j.ympev.2016.11.007. PMID 27845203. Piraino S, Boero F, ... those that propose aging to be caused by specific molecular changes occurring over time. One theory was proposed by George C. ...
Patton JG, Mayer SA, Tempst P, Nadal-Ginard B (July 1991). "Characterization and molecular cloning of polypyrimidine tract- ... Patton JG, Porro EB, Galceran J, Tempst P, Nadal-Ginard B (March 1993). "Cloning and characterization of PSF, a novel pre-mRNA ... Patton JG, Porro EB, Galceran J, Tempst P, Nadal-Ginard B (March 1993). "Cloning and characterization of PSF, a novel pre-mRNA ... Journal of Molecular Biology. 298 (3): 395-405. doi:10.1006/jmbi.2000.3687. PMID 10772858. Conte MR, Grüne T, Ghuman J, Kelly G ...
"Entrez Gene: CD34 CD34 molecule". Simmons DL, Satterthwaite AB, Tenen DG, Seed B (January 1992). "Molecular cloning of a cDNA ... Simmons DL, Satterthwaite AB, Tenen DG, Seed B (January 1992). "Molecular cloning of a cDNA encoding CD34, a sialomucin of ... April 1997). "The characterization, molecular cloning, and expression of a novel hematopoietic cell antigen from CD34+ human ... October 2009). "The molecular basis of vascular lumen formation in the developing mouse aorta". Developmental Cell. 17 (4): 505 ...
The molecular weight of the protein is 76.5 kilodaltons, and the isoelectric point is 5.47.The gene has 6 transcript splice ... Cloning and functional analysis of cDNAs with open reading frames for 300 previously undefined genes expressed in CD34+ ...
... DNA. 1986 Aug;5(4):315-24. doi: 10.1089/dna.1986.5.315. ... has been cloned and sequenced (Oesch et al., 1985). Using that hamster PrP cDNA, we screened a human retina cDNA library and ... sequenced the cDNA clone with the longest hybridizing insert. This insert was found to contain a long open reading frame (ORF) ... has been cloned and sequenced (Oesch et … ...
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Molecular Characterization of Campylobacter jejuni Clones: A Basis for Epidemiologic Investigation Kate E. Dingle*, Frances M. ... Molecular Characterization of Campylobacter jejuni Clones: A Basis for Epidemiologic Investigation. ...
Chapter 9 Molecular Cloning, Expression, Purification and Im... By Shumaila Nida Muhammad Hanif, Rajaa Al-Attiyah and... ... Chapter 8 Molecular Cloning, Characterization, Expression An... By Gloria Esteso, Ángeles Jiménez-Marín, Gema Sanz... ... Molecular Cloning Selected Applications in Medicine and Biology Edited by Gregory G. Brown ... Molecular Cloning Selected Applications in Medicine and Biology Edited by Gregory G. Brown ...
1991) Molecular cloning of a gene encoding the histamine H2 receptor. Proc Natl Acad Sci USA 88:429-433. ... The GPCR97 clone was used to probe a human thalamus library, which resulted in the isolation of a full-length clone encoding a ... Through the molecular cloning of H1 and H2, these receptors were proven to belong to the superfamily of G protein-coupled ... the histamine H3 receptor has been the target of numerous cloning and purification attempts, yet its molecular identity has ...
Molecular cloning and characterization of a ripening-induced polygalacturonase related to papaya fruit softening. Plant ... Molecular cloning and characterization of a ripening-induced polygalacturonase related to papaya fruit softening Full text ...
The identification and molecular cloning of bioactive peptides from the venom of scorpion Androctonus Australis ...
The cloning of the mouse galanin gene should allow elucidation of the regulatory characteristics of its promoter and facilitate ... a genomic clone was isolated covering the entire mouse preprogalanin gene. The mouse gene has an exon:intron organisation very ... Molecular cloning and characterisation of the mouse preprogalanin gene. Abstract. Using a probe obtained by PCR amplification ... The cloning of the mouse galanin gene should allow elucidation of the regulatory characteristics of its promoter and facilitate ...
Molecular Cloning and Expression of Nucleoprotein gene (NPs) of tomato spotted wilt virus in E.coli ... Molecular Cloning and Expression of the Coat Protein Gene of Plum Pox Virus El-Amar Strain in E.coli. Cloning, exprision and ... Molecular Cloning and Expression of Nucleoprotein gene (NPs) of tomato spotted wilt virus in E.coli. E. T. Abdelsalam, Hala M. ... amplicon was cloned, and expressed into the pBAD-C expression vector in E.coli. The NPs fusion protein was induced using 0.002 ...
TRP: Molecular cloning, characterization and catalytic mechanism ...
Molecular cloning, nucleotide sequence and characteristics of a xylanase gene (xynA) from Ruminococcus albus 7. In: Animal ... Molecular cloning, nucleotide sequence and characteristics of a xylanase gene (xynA) from Ruminococcus albus 7. Animal Science ... Molecular cloning, nucleotide sequence and characteristics of a xylanase gene (xynA) from Ruminococcus albus 7. / Nakamura, ... title = "Molecular cloning, nucleotide sequence and characteristics of a xylanase gene (xynA) from Ruminococcus albus 7", ...
Molecular cloning of the fimbrial subunit gene from a benign type B isolate ofBacteroides nodosus. FEMS Microbiology Letters. ... Molecular cloning of the fimbrial subunit gene from a benign type B isolate ofBacteroides nodosus. / Boulos, Sherif; Rood, ... Boulos, S., & Rood, J. I. (1986). Molecular cloning of the fimbrial subunit gene from a benign type B isolate ofBacteroides ... Boulos, Sherif ; Rood, Julian I. / Molecular cloning of the fimbrial subunit gene from a benign type B isolate ofBacteroides ...
Reference Plasmid pHXB2_D is an HIV-1 Molecular Clone that Exhibits Identical LTRs and a Single Integration Site Indicative of ...
Molecular cloning of a novel kinin-forming enzyme and its tissue expression in the human cardiovascular organ. Research Project ... Kallikrein / Kinin / cloning / human tissues. Research Abstract. Abstract. We previously purified a novel kallikrein-like ... So far, we have not cloned a kallikrein gene from the human heart and neutrophil cDNA library by using this transcript as a ... To clarity whether this novel kallikrein-like enzyme is present in human tissue, we tried to clone this enzyme. In the present ...
Molecular cloning and sequence of Sparus aurata skeletal myosin light chains expressed in white muscle: developmental ... Amino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, DNA, Complementary, Gene Expression Regulation, Developmental ... Two full-length cDNA clones encoding the skeletal myosin light chain 2 (MLC2; 1452bp) and myosin light chain 3 (MLC3; 972bp) ... Gene Library, Molecular Sequence Data, Muscle, Skeletal, Myosin Light Chains, Perciformes, Phylogeny, Reverse Transcriptase ...
Molecular cloning and functional expression of rabbit α2-antiplasmin Academic Article ...
We have therefore decided to construct an STLV-3 molecular clone. We generated a full-length STLV-3 proviral clone (8,891 bp) ... All together, our data demonstrate that the STLV-3 molecular clone is functional and infectious. This clone will give us a ... This STLV-3 molecular clone was then transfected into 293T cells. Reverse transcriptase PCR experiments followed by sequence ... Construction and characterization of a full-length infectious simian T-cell lymphotropic virus type 3 molecular clone. ...
Molecular cloning and analysis of the mouse homologue of the tumor-associated mucin, MUC1, reveals conservation of potential O- ... Dive into the research topics of Molecular cloning and analysis of the mouse homologue of the tumor-associated mucin, MUC1, ...
BioConstructor Molecular Cloning Software Lenti vector viral particles from E. coli plasmids with Puro ... BioConstructor Molecular Cloning Software. Lenti vector viral particles from E. coli plasmids with Puro ...
Using mutagenesis and molecular cloning techniques to identify key residues within the molecular recognition site of the ... Using mutagenesis and molecular cloning techniques to identify key residues within the molecular recognition site of the ... Successful cloning to create mutant GusB proteins could not be achieved however, sequence analysis of a previous plasmid pE349 ... This led to questions as to whether this amino acid change was indeed part of the molecular recognition site. All steps ...
Molecular cloning and sequence analysis of the Chlamydomonas gene coding for radial spoke protein 3: flagellar mutation pf-14 ... B D Williams, M A Velleca, A M Curry, J L Rosenbaum; Molecular cloning and sequence analysis of the Chlamydomonas gene coding ... Molecular cloning and expression of flagellar radial spoke and dynein genes of Chlamydomonas. ...
GFP Vector Set (plasmid vectors for molecular cloning). Molecular cloning often benefits from optimizing the vector used for ...
Molecular cloning of doublesex genes of four cladocera (water flea) species. BMC genomics. 2013 Apr 10;14(1):239. doi: 10.1186/ ... Molecular cloning of doublesex genes of four cladocera (water flea) species. In: BMC genomics. 2013 ; Vol. 14, No. 1. ... Molecular cloning of doublesex genes of four cladocera (water flea) species. Kenji Toyota, Yasuhiko Kato, Masaru Sato, Naomi ... Molecular cloning of doublesex genes of four cladocera (water flea) species. / Toyota, Kenji; Kato, Yasuhiko; Sato, Masaru et ...
Based on SSR analysis, linkage mapping and genome database survey, we cloned a candidate gene designated as GmF3G2″Gt in the ... The second objective was to clone the candidate genes and to verify their function. Recombinant inbred lines (RILs) were ... molecular linkage group C2 (chromosome 6). The open reading frame of GmF3G2″Gt is 1380 bp long encoding 459 amino acids with ... From: Linkage mapping, molecular cloning and functional analysis of soybean gene Fg3 encoding flavonol 3-O-glucoside/ ...
Molecular Cloning and Chromosomal Localization of the Human T-Cell Receptor ζ Chain: Distinction from the Molecular CD3 Complex ... "Molecular Cloning and Chromosomal Localization of the Human T-Cell Receptor ζ Chain: Distinction from the Molecular CD3 Complex ...
Molecular" by people in this website by year, and whether "Cloning, Molecular" was a major or minor topic of these publications ... "Cloning, Molecular" is a descriptor in the National Library of Medicines controlled vocabulary thesaurus, MeSH (Medical ... Below are the most recent publications written about "Cloning, Molecular" by people in Profiles. ... Below are MeSH descriptors whose meaning is more general than "Cloning, Molecular". ...
Its hybrid origin needs to be confirmed using molecular technology. We investigated the origin of S. longifolium based on 10 ... Amplification, sequencing and cloning. We sequenced one chloroplast DNA fragment trnH-psbA12 and six nuclear loci developed ... which did not provide directly molecular evidence to clarify the origin of S. longifolium6. Further molecular studies are still ... Yu, Y., Li, F., Belyakov, E.A. et al. Molecular confirmation of the hybrid origin of Sparganium longifolium (Typhaceae). Sci ...
  • Over the years, understanding the biology and pathogenesis of HTLV-1 and HTLV-2 has been widely improved by the creation of molecular clones. (
  • NEB is the Doble Lab's source for restriction and cloning enzymes and a wide variety of molecular biology kits. (
  • molecular cloning, fourth edition, by the molecular cloning a laboratory sambrook, Download Molecular Cloning A Laboratory Manual 2nd Edition Pdf posttransfection analysis of cells flow cytometry fluorometry laserscanning molecular imaging luminometry microscopyprinciples and techniques of biochemistry and molecular biology seventh edition edited by keith wilson and john walker this new edition of the bestselling textbookbiosafety module resource book a introduction to. (
  • Background Vectors based mostly on plant viruses are essential instruments for purposeful genomics, mobile biology, plant genome engineering and molecular farming. (
  • The invention of PCR was revolutionary to molecular biology . (
  • These probes are used in nucleic acid hybridization, in situ hybridization and other molecular biology procedures. (
  • THESE days we can't escape from molecular biology-neither its achievements nor its hype. (
  • Molecular biology is growing so rapidly that the half-life of papers is but a few months, and students and postdocs rarely read or refer to anything published more than three years ago. (
  • In the name of molecular biology, they have destroyed what is to him the essence of our science-respect for and understanding of living processes. (
  • It was with relief, then, that I turned to Morange's well-researched and clearly written A History of Molecular Biology, which appeared in French in 1994 (elegantly translated by Matthew Cobb). (
  • Animal Cloning, Updated Edition discusses all aspects of this new biology, including the scientific, ethical, and legal issues. (
  • Joseph Panno, Ph.D. , holds a degree in biology from Simon Fraser University in British Columbia and specializes in molecular biology and physiology. (
  • Leverage high-throughput molecular biology to build large sets of DNA reagents for testing in cell culture. (
  • Dr. Bell has long-standing interest in the molecular biology and genetics of diabetes. (
  • and (i i) to develop (epi)genomic powerful molecular and/or cell biology and close col aboration to create synergies methodologies, profiling strategies, and functional genomics tools, recent progress and better exploit and further expand bioinformatics tools and resources that in understanding of the cancer (epi) unique research tools and expertise. (
  • Molecular cloning and characterization of a ripeni. (
  • Katoh M. Molecular cloning and characterization of human WNT3. (
  • STs, serotypes and PorA VR types as found in Hajj-related N. characterization of invasive isolated in Brazil from 1990 approaches meningitidis serogroup W135 clone. (
  • Using a probe obtained by PCR amplification from mouse genomic DNA, a genomic clone was isolated covering the entire mouse preprogalanin gene. (
  • The cloning of the mouse galanin gene should allow elucidation of the regulatory characteristics of its promoter and facilitate transgenic approaches to the analysis of galanin gene function in this species. (
  • A 2.6‐kb DNA fragment encoding a xylanase gene (xynA) was cloned from the rumen hemicellulolytic bacterium Ruminococcus albus 7. (
  • The fimbrial subunit gene from the benign type BBacteroides nodosus isolate AC/6 was cloned into theSphI site of the multicopy vector plasmid pUC19. (
  • Boulos, S & Rood, JI 1986, ' Molecular cloning of the fimbrial subunit gene from a benign type B isolate ofBacteroides nodosus ', FEMS Microbiology Letters , vol. 33, no. 1, pp. 73-78. (
  • So far, we have not cloned a kallikrein gene from the human heart and neutrophil cDNA library by using this transcript as a target DNA. (
  • Molecular cloning and sequence analysis of the Chlamydomonas gene coding for radial spoke protein 3: flagellar mutation pf-14 is an ochre allele. (
  • Molecular phylogenetic analysis suggested that the dsx gene duplication likely occurred prior to the divergence of these cladoceran species, because that of the giant tiger prawn Penaeus monodon is rooted ancestrally to both DSX1 and DSX2 of cladocerans. (
  • Cloning, expression, purification and biophysical analysis of two putative halogenases from the glycopeptide A47,934 gene cluster of Streptomyces toyocaensis. (
  • IMSEAR at SEARO: Molecular cloning of coat protein gene of cucumber mosaic virus, CMV-U strain. (
  • Raj SK, Leiser RM, Plobner L. Molecular cloning of coat protein gene of cucumber mosaic virus, CMV-U strain. (
  • ID13 cDNA library was screened with human hsp70 gene as a probe, and a positive clone was obtained. (
  • The sequence of cloned mouse hsp70 shows great homology (60-97\%) with other mouse hsp 70 genes and somewhat less homology (50\%) with E. coli hsp 70 gene (dnaK). (
  • The great homology (97\%) with already cloned mouse heat shock cognate 70 kDa protein (hsc 70) make us to think that the cloned gene is constitutively expressed at normal temperature. (
  • Molecular cloning, sequence analysis and transcriptional activity determination of cytochrome P450 gene CYP18A1 in the silkworm, Bombyx mori[J]., 2008, 51(3): 237-245. (
  • Cloning of ferritin gene AmFer3HCH and its response to temperature stress in the Italian honeybee, Apis mellifera ligust ica (Hymenoptera: Apidae) [J]. , 2018, 61(9): 1029-1039. (
  • Molecular cloning of a serine protease gene DdSP and its response to temperature stress in Galeruca daurica (Coleoptera: Chrysomelidae) [J]. , 2018, 61(7): 761-770. (
  • Cloning, prokaryotic expression and tissue expression profiling of an odorant binding protein gene BminOBP 25 from Bactrocera minax (Diptera: Tephritidae) [J]. , 2018, 61(5): 537-545. (
  • 2. Nuclear transfer is a technique used to duplicate genetic material by creating an embryo through the transfer and fusion of a diploid cell in an enucleated female oocyte.2 Cloning has a broader meaning than nuclear transfer as it also involves gene replication and natural or induced embryo splitting (see Annex 1). (
  • Roelink H, Wang J, Black DM, Solomon E, Nusse R. Molecular cloning and chromosomal localization to 17q21 of the human WNT3 gene. (
  • The molecular sequence of this clone aligns with the gene accession number as a point of reference only. (
  • Hierdie is die eerste epidemiologiese studie by Tygerberg hospitaal wat prospektiewe kliniese data van pasiënte met S. aureus bakteremie saam met spa tipering en aantoning van die mecA en pvl gene in 'n multipleks PKR insluit. (
  • Molecular cloning of the gene showed that HER2, Neu, and ErbB-2 are all encoded by the same orthologs. (
  • Douglas Prasher Douglas Prasher is the first one who cloned this gene. (
  • So, he cloned this gene. (
  • If you dont see any interesting for you, use our search form on bottom v. In order to facilitate the topics of restriction enzyme digestion and the generation of compatible ends in the process of gene cloning, an inclass activity was designed. (
  • 7468 /organism="Norovirus GII.6" /mol_type="genomic RNA" /isolation_source="stool" /host="Homo sapiens" /db_xref="taxon:499193" /clone="V4B" /environmental_sample /country="Venezuela" /collection_date="2015" /metagenome_source="gut metagenome" /note="metagenomic" gene 5. (
  • For the past 10 years, the histamine H 3 receptor has been the target of numerous cloning and purification attempts, yet its molecular identity has remained an enigma. (
  • Purification, molecular cloning and heterologous expression of a glutathione S-transferase involved in insecticide resistance from the rice brown planthopper, Nilaparvata lugens. (
  • The observed distribution of resistance plasmids and β-lactamase genes in several clones indicates a high degree of horizontal transfer. (
  • Now, interestingly enough is these genes (these antimicrobial resistant genes), they can find their way onto certain clones. (
  • And these clones, when the genes reach them, they act as hoarders and spreaders of these antimicrobial genes. (
  • As cepas de P. aeruginosa foram identificadas por MALDI-TOF MS, submetidas a testes de susceptibilidade aos antimicrobianos, identificação de genes de virulência através de PCR e tipagem molecular através de PFGE. (
  • P. aeruginosa strains were identified by MALDI-TOF MS, subjected to antimicrobial susceptibility testing, identification of virulence genes by PCR and molecular typing by PFGE. (
  • 5. In 2001, France and Germany requested the United Nations General Assembly to develop international conventions on human reproductive cloning, therapeutic cloning and research on stem cells. (
  • 2001. Molecular dynamics studies of sequence-directed curvature in bending locus of Trypanosome kinetoplast DNA . (
  • 2001. Molecular dynamics of minimal B-DNA . (
  • Prevalence of Major Methicillin-Resistant Staphylococcus aureus Clones in Korea Between 2001 and 2008. (
  • A successful PCR amplification of the virus-specific DNA fragment (777 bp) amplicon was cloned, and expressed into the pBAD-C expression vector in E.coli. (
  • Double-stranded cDNA was cloned in PRT103 vector at Xho1/Kpn1 site and about 1kb insert obtained. (
  • Cloning of P5 and P7 Illumina adaptor sequences into the vector backbone allows direct Sequencing of the viral cassette. (
  • Vector for standard cloning and efficient in vitro RNA synthesis. (
  • Successful cloning to create mutant GusB proteins could not be achieved however, sequence analysis of a previous plasmid pE349 encoding a mutant GusB found an unexpected amino acid mutation at position 218 of the glucuronide membrane transporter. (
  • To screen for probable target(s), we introduced the ASKA pooled-plasmid library into Escherichia coli W3110 imp4213 and enriched the library for resistant clones with increasing concentrations of xanthorrhizol. (
  • The plasmid pSP65 which has cloned the complete HBV sequence, was amplified by transformation of coli (DH5). (
  • WHA50.37 of 1997 argues that human cloning is ethically unacceptable and contrary to human integrity and morality. (
  • Infec- for each strain were de novo assembled by using CLC tions were mostly associated with the pandemic clone Genomics Workbench version 7.5.1 (QIAGEN, Valencia, throughout 1997 and 1998 and then with a less clear pat- CA, USA). (
  • We used Spectral Karyoryping (SKY), mapping with fluorescently labeled genomic clones (FISH), comparative genomic hybridization (CGH), expression array, real time polymerase chain reaction and Western blot to analyze 15 primary adenocarcinoma and 9 pairs of high and low invasive cell cultures to detect molecular changes. (
  • Genomic Analysis of Carbapenem-Resistant Pseudomonas aeruginosa Isolated From Urban Rivers Confirms Spread of Clone Sequence Type 277 Carrying Broad Resistome and Virulome Beyond the Hospital. (
  • In the current paper, we have isolated bovine and rat cDNA clones encoding PPT. (
  • These recombinant clones will prove useful for the construction of a multivalent recombinant vaccine for the control of ovine footrot. (
  • The recombinant clones were sequenced, which had the same sequence as the predictive contig. (
  • Our study expanded the previous investigation to include cases from 2011 to 2014 and to perform Multi-Virulence-Locus Sequence Typing (MVLST) on isolates in Cluster11/ST38 to better understand their epidemiology and possibly identify a common source outbreak clone. (
  • The molecular epidemiology of hospital acquired (HA), health-care associated (HCA) and community acquired (CA) S. aureus bacteraemic strains at this hospital was examined. (
  • is also a great information resource for molecular cloning, genome editing, fluorescent proteins and much more. (
  • Small peptides, especially those derived from natural proteins as inhibitory peptide aptamers (iPAs), can produce highly effective and selective blockade of specific nociceptive molecular pathways to reduce pain with minimal off-target effects. (
  • In nature, many organisms produce clones through asexual reproduction.Cloning in biotechnology refers to the process of creating clones of MOLECULAR CLONING A LABORATORY MANUAL 2ND EDITION PDF READ Molecular Cloning A Laboratory Manual 2nd Edition pdf. (
  • Molecular cloning : a laboratory manual / T. Maniatis, E. F. Fritsch, J. Sambrook. (
  • molecular cloning a laboratory manual second edition pdf no other manual has been so popular, or so influential. (
  • Molecular Cloning A Laboratory Manual Second Edition. (
  • T. Maniatis, Harvard, Great Molecular Cloning A Laboratory Manual 2nd Edition. (
  • molecular cloning laboratory manual second edition Fri, 21 Dec 2018 17:09:00 GMT molecular cloning laboratory manual second pdf - Cloning is the process of producing genetically identical individuals of an organism either naturally or artificially. (
  • Molecular biologists use PCR to cloning DNA. (
  • He argues that molecular biologists suffer from a delusion (he calls it "delirium genetica") that the unfolding of living processes in the four dimensions of space and time can be read off the one-dimensional strand of DNA. (
  • SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files. (
  • We generated a full-length STLV-3 proviral clone (8,891 bp) by PCR amplification of overlapping fragments. (
  • Researchers in the Human Genome Project use PCR to look for markers in cloned DNA segments and to order DNA fragments in libraries. (
  • The fragments were cloned and some clones were size-selectd and sequenced. (
  • Using E. coli as a model organism, site-directed mutagenesis and cloning techniques employed in the laboratory were used to create six residual amino acid changes within the Glucuronide Transporter Protein. (
  • A clone is an organism that is a genetic copy of an existing one. (
  • Initially illnesses were reported only in the genome sequencing of clinical and environmental samples from the outbreak revealed a new V. parahaemolyticus northern cities of Peru (Cajamarca, Chiclayo, and Piura), clone. (
  • Whole-genome sequence contigs clone ST-3, which also originated in Asia ( 8 , 12 ). (
  • It circumvents cell heterogeneity, a consequence of Cas9 genome editing, by scoring single cell derived clones individually. (
  • By substituting amino acids to others with different charges and polarity, it was expected that protein folding mechanisms and the molecular recognition site would be disrupted. (
  • Recently, we are examining molecular mechanisms by which prostanoids interact with other lipid signaling systems to protect the vascular system and maintain health. (
  • Some of the more interesting cellular and molecular mechanisms are reviewed briefly in this article. (
  • Molecular Cloning and Chromosomal Localization of the Human T-Cell Rec" by Allan M. Weissman, Damon Hou et al. (
  • 1995) Cloning, expression, and chromosomal localization of human uridine nucleotide receptors. (
  • This STLV-3 molecular clone was then transfected into 293T cells. (
  • Transient transfection of 293T cells and of 293T-long terminal repeat-green fluorescent protein cells with the STLV-3 clone promoted syncytium formation, a hallmark of PTLV Env expression, as well as the appearance of fluorescent cells, also demonstrating that the Tax3 protein was expressed. (
  • Nature has been cloning animals, cells, and molecules for millions of years. (
  • American molecular biologist Kary Mullis developed the techniques of PCR in the 1970s. (
  • In 1996 when Ian Wilmut, a British biologist, cloned a sheep named Dolly, the reaction was dramatically different. (
  • molecular cloning, fourth edition, by the molecular cloning a laboratory sambrook molecular cloning a laboratory pdf molecular cloning refers to the process of making multiple molecules. (
  • FiveEscherichia coli recombinants that were positive in a colony immunoassay were shown, by Western transfer analysis, to produce an immunologically cross-reacting protein of identical molecular size to fimbrial subunits prepared fromB. (
  • Therefore, molecular identification is an essential method for the study of natural hybridization in Sparganium . (
  • The full-length open reading frame of the canine REIC/Dkk-3 homologue was cloned and the tissue distribution of REIC/Dkk-3 mRNA was determined, along with the subcellular localisation of the REIC/Dkk-3 protein in canine cancer cell lines. (
  • Before molecular analysis, the genotypes and subgenotypes associated with these outbreaks. (
  • 2011) Molecular cloning and analysis of the UDP-glucose pyrophosphorylase in Streptococcus equi subsp. (
  • Clonal cluster groups of S. aureus isolates were obtained by BURP analysis and compared to international important clones. (
  • The production of IgE is tightly regulated and involves a complex network of cellular and molecular signals. (
  • Here we will give a brief overview of the roles of IgE in allergic pathophysiology, and the molecular and cellular factors that ultimately regulate IgE production and Th2 expansion. (
  • We're trying to understand the molecular, cellular nature and the circuitry arrangement of how nervous system works. (
  • Here we present CRISPR-UMI (Unique Molecular Identifier), a single cell tracing approach, providing a robust screening method that can detect, and thus overcome, cellular heterogeneity and clonal outliers. (
  • The positive clone was subcloned into pUC19 and the precise restriction map was obtained. (
  • exposures and lifestyle alter critical advances as well as the uniqueness and These developments have also led to molecular pathways and promote cancer strengths of IARC. (
  • A cDNA clone of Clonorchis sinensis (CsFtn), 565 bp long, encoded a putative polypeptide of 166 amino acids. (
  • The GPCR97 clone was used to probe a human thalamus library, which resulted in the isolation of a full-length clone encoding a putative G protein-coupled receptor. (
  • Documentation of laboratory-confirmed SARS-CoV-2 infection, as determined by a molecular (nucleic acid) or antigen test from any respiratory tract specimen (e.g., oropharyngeal, NP, or nasal swab, or saliva) collected ââ? (
  • Cloning, Molecular" is a descriptor in the National Library of Medicine's controlled vocabulary thesaurus, MeSH (Medical Subject Headings) . (
  • All together, our data demonstrate that the STLV-3 molecular clone is functional and infectious. (
  • We describe an infectious molecular clone of a Japanese genotype lb strain of hepatitis C virus (HCV-N). The molecularly cloned sequence of HCV- N was compared with alignments of other HCV sequences, leading to the identification of 15 unique, nonconservative amino acid substitutions within the HCV-N open reading frame (ORF). (
  • The HCV- N clone is the first infectious molecular clone of HCV that is comprised entirely of genotype lb sequence, and it contains an ORF sequence that is significantly divergent from that of a previously described genotype la/lb chimera. (
  • Klonale groepe (spa-CC) van MRSA en MSSA isolate is deur BURP analise verkry, en vergelyk met internasionaal belangrike klone. (
  • Laastens, oorspronklike en daaropvolgende herhaal isolate is gekollekteer om moontlike organisme- faktore geassosieerd met persisterende en herhalende bakteremiese episodes te analiseer. (
  • The isolated cDNA (1.1 kb) contained an 858 bp open reading frame encoding a 286 amino acid polypeptide with molecular weight of 33439. (
  • First is mobile genetic elements, and then they find their way onto these special or successful clones and then they tend to keep. (
  • The MRSA population in the Philippines comprised a limited number of genetic clones, including several international epidemic clones, such as CC30-spa-t019-SCCmec-IV-PVL+, CC5-SCCmec-typeIV and ST239-spa-t030-SCCmec-typeIII. (
  • We showed independent acquisition of resistance to sulfamethoxazole/trimethoprim in various locations and genetic clones but mostly in paediatric patients with invasive infections. (
  • This can be determined using the Revised International Prognostic Scoring System (IPSS-R) in addition to evaluation of a patient's performance status, symptoms, care goals, cytopenias, and molecular genetic testing (eg, for 5q31 deletion or SF3B1 mutations). (
  • 3. Media reports on nuclear transfer are usually about one form, reproductive nuclear transfer, also known as reproductive cloning of human beings . (
  • Beginning chapters discuss cloning within the context of a natural process that many animals use as a survival strategy, followed by the historical development of the nuclear transfer procedure, the cloning of Dolly the sheep, the medical applications of cloning technology, plant clones, and much more. (
  • We found that radial clones do not appear to be fate-restricted, but instead individual clones are composed of a random sampling of the transcriptomic cell types present in a particular cortical area. (
  • From cloned sheep to sterile corn, we are surrounded by more or less informed debates about what its effects will be. (
  • Resistência antimicrobiana e tipagem molecular de pseudomonas aeruginosa isoladas de feri. (
  • We have therefore decided to construct an STLV-3 molecular clone. (
  • To cure molecular biology's illness, he argues for a form of neo-Lamarckism, drawing on ideas ranging from Stuart Kauffman's self-organising networks of mutually-catalysing reactions, James Lovelock's Gaia hypothesis, to the postmodernist philosophy of Jacques Derrida. (
  • 1985). Using that hamster PrP cDNA, we screened a human retina cDNA library and sequenced the cDNA clone with the longest hybridizing insert. (
  • 2. Over the years, the international community has tried without success to build a consensus on an international convention against the reproductive cloning of human beings. (
  • General Assembly the adoption of a declaration on human cloning by which Member States were called upon to prohibit all forms of human cloning inasmuch as they are incompatible with human dignity and the protection of human life. (
  • 3. Creating awareness among ministries of health in the African Region will provide them with critical and relevant information on the reproductive cloning of human beings and its implications to the health status of the general population. (
  • 7. The WHO Regional Committee for Africa is invited to review this document for information and guidance concerning reproductive cloning of human beings. (
  • WHA50.37, which states "the use of cloning for the replication of human individuals is ethically unacceptable and contrary to human integrity and morality. (
  • To elucidate the role of Nanos in an amphibian species, we cloned Nanos3 cDNA from the testis of the R. rugosa frog. (
  • In this study, Jarid2b was cloned and characterized in Nile tilapia , which was highly conserved among the analyzed fish species. (