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
Molecular cloning[edit]. Most strains used by molecular biologists are derivatives of E. coli K-12, and possess both Dam and ... Gardiner-Garden M, Frommer M (July 1987). "CpG islands in vertebrate genomes". Journal of Molecular Biology. 196 (2): 261-82. ... Molecular break light assay for DNA adenine methyltransferase activity - an assay that relies on the specificity of the ... Ying and Li-Byarlay (2015). "Physiological and Molecular Mechanisms of Nutrition in Honey Bees". Advances in Insect Physiology ...
Gene isolation and cloning[edit]. Main article: Molecular cloning. The next step is to isolate the candidate gene. The cell ... Koh H, Kwon S, Thomson M (26 August 2015). Current Technologies in Plant Molecular Breeding: A Guide Book of Plant Molecular ... Mutagenesis (molecular biology technique). References[edit]. *^ "Terms and Acronyms". U.S. Environmental Protection Agency ... "Isolating, Cloning, and Sequencing DNA (4th ed.). New York: Garland Science.. *^ Kaufman RI, Nixon BT (July 1996). "Use of PCR ...
Molecular cloning and expression". Biochem. J. 329 ( Pt 2) (Pt 2): 275-82. PMC 1219041 . PMID 9425109.. ... Molecular function. • zinc ion binding. • peptidase activity. • aminopeptidase activity. • protein binding. • hydrolase ...
"Molecular and Cellular Biology. 25 (14): 5846-58. doi:10.1128/MCB.25.14.5846-5858.2005. PMC 1168807. PMID 15988002.. ... Cloning and Stem Cells. 11 (3): 427-35. doi:10.1089/clo.2009.0024. PMID 19751112.. ... "International Journal of Molecular Sciences. 20 (14): 3563. doi:10.3390/ijms20143563. PMC 6679312. PMID 31330871.. ... "International Journal of Molecular Sciences. 21 (7): 2598. doi:10.3390/ijms21072598. PMC 7177281. PMID 32283613.. ...
Molecular cloning and chromosomal localization". Advances in Experimental Medicine and Biology. 414: 253-260. doi:10.1007/978-1 ... Saronwala, A.; Tournay, A.; Gargus, J. J. "Genetic inborn error of metabolism provides a unique window into molecular ... "Two exon-skipping mutations as the molecular basis of succinic semialdehyde dehydrogenase deficiency (4-hydroxybutyric ...
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 0-8053-9592-X. Russell, David W.; Sambrook, Joseph (2001). Molecular cloning: a laboratory manual. Cold Spring Harbor, N.Y ... This is particularly determined by the number of clones needed to have in a library. The number of clones to get a sampling of ...
Molecular cloning is one of the most commonly used procedures in molecular biology. A gene of interest may be inserted into a ... "Chapter 1". Molecular Cloning - A Laboratory Manual. 1 (3rd ed.). p. 1.27. ISBN 978-0-87969-577-4.. ... "White and green screening with circular polymerase extension cloning for easy and reliable cloning". Protein Science. 22 (6): ... The plasmid carries within the lacZα sequence an internal multiple cloning site (MCS). This MCS within the lacZα sequence can ...
Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY. ...
"Future and applications of cloning". Methods Mol. Biol. Methods in Molecular Biology. 348: 319-32. doi:10.1007/978-1-59745-154 ... The attempt to clone a banteng bull was more successful, as were the attempts to clone mouflon (a form of wild sheep), both ... The cell used as the donor for the cloning of Dolly was taken from a mammary gland, and the production of a healthy clone ... Even though Dolly was not the first animal cloned, she received media attention because she was the first cloned from an adult ...
"European Molecular Biology Laboratory - Hamburg. Retrieved 2008-06-07.. *^ Russell DW, Sambrook J (2001). Molecular cloning: a ... and they are a vital tool in molecular cloning.[8][9][10] ... Enzymes of Molecular Biology. Methods of Molecular Biology. 16 ... "Microbiology and Molecular Biology Reviews. 64 (2): 412-34. doi:10.1128/MMBR.64.2.412-434.2000. PMC 98998. PMID 10839821.. ... this characteristic is widely used to perform in-vitro cloning techniques such as Golden Gate cloning. These enzymes may ...
Sambrook, Joseph; Russell, David W. (2001). "Commonly Used Techniques in Molecular Cloning". Molecular Cloning. 3. Li, Richard ... Phenol-chloroform extraction is a liquid-liquid extraction technique in molecular biology used to separate nucleic acids from ...
Ohara, O. (2009). "ORFeome Cloning". Reverse Chemical Genetics. Methods in Molecular Biology. 577. pp. 3-9. doi:10.1007/978-1- ... 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 ...
Tomato as a model system: I. Genetic and physical mapping of jointless". MGG Molecular & General Genetics. 242 (6). doi:10.1007 ... Tomatoes have been used as a model in map-based cloning, where transgenic plants must be created to prove that a gene has been ... Foolad, M. R. (2007). "Current Status Of Breeding Tomatoes For Salt And Drought Tolerance". Advances in Molecular Breeding ... Wing, R.; Zhang, H. B.; Tanksley, S. (1994). "Map-based cloning in crop plants. ...
Russell MW, Kemp P, Wang L, Brody LC, Izumo S (December 1998). "Molecular cloning of the human HAND2 gene". Biochimica et ... Molecular function. • DNA binding. • sequence-specific DNA binding. • RNA polymerase II regulatory region sequence-specific DNA ... Srivastava D, Gottlieb PD, Olson EN (2003). "Molecular mechanisms of ventricular hypoplasia". Cold Spring Harbor Symposia on ... Srivastava D (1999). "HAND proteins: molecular mediators of cardiac development and congenital heart disease". Trends in ...
"Molecular cloning of the human eosinophil peroxidase. Evidence for the existence of a peroxidase multigene family". J. Exp. Med ... Petrides, PE (September 1998). "Molecular genetics of peroxidase deficiency". Journal of molecular medicine (Berlin, Germany). ... Molecular weight: 57 kDa (heavy chain), 11 kDa (light chain) (predicted); 52 kDa, 15 kDa (observed) Isoelectric point pI = ... Lanza, F (September 1998). "Clinical manifestation of myeloperoxidase deficiency". Journal of molecular medicine (Berlin, ...
Gene isolation and cloningEdit. Main article: Molecular cloning. The next step is to isolate the candidate gene. The cell ... Koh H, Kwon S, Thomson M (26 August 2015). Current Technologies in Plant Molecular Breeding: A Guide Book of Plant Molecular ... "Isolating, Cloning, and Sequencing DNA (4th ed.). New York: Garland Science.. *^ Kaufman RI, Nixon BT (July 1996). "Use of PCR ... "Microbiology and Molecular Biology Reviews. 67 (1): 16-37, table of contents. doi:10.1128/MMBR.67.1.16-37.2003. PMC 150518. ...
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. ...
"Molecular cloning of a functional human galanin receptor". Proceedings of the National Academy of Sciences of the United States ... Molecular function. • G-protein coupled receptor activity. • neuropeptide binding. • signal transducer activity. • peptide ... Lorimer DD, Benya RV (May 1996). "Cloning and quantification of galanin-1 receptor expression by mucosal cells lining the human ... Lorimer DD, Matkowskj K, Benya RV (Dec 1997). "Cloning, chromosomal location, and transcriptional regulation of the human ...
"Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 1823 (9): 1426-33. doi:10.1016/j.bbamcr.2012.03.004. PMC 3718258 ... February 2000). "Positional cloning of zebrafish ferroportin1 identifies a conserved vertebrate iron exporter". Nature. 403 ( ... "DNA cloning using in vitro site-specific recombination". Genome Research. 10 (11): 1788-95. doi:10.1101/gr.143000. PMC 310948 ... Molecular Diagnosis. 6 (4): 347. doi:10.1054/modi.2001.0060347. PMID 11774199.. ...
... properties and molecular cloning". Proc Natl Acad Sci U S A. 92 (19): 8695-9. doi:10.1073/pnas.92.19.8695. PMC 41033 . PMID ... Molecular function. • transferase activity. • enoyl-[acyl-carrier-protein reductase (NADPH, A-specific) activity]. • 3- ... Stevens L, Price NC (1999). Fundamentals of enzymology: the cell and molecular biology of catalytic proteins. Oxford [ ... Asturias FJ, Chadick JZ, Cheung IK, Stark H, Witkowski A, Joshi AK, Smith S (March 2005). "Structure and molecular organization ...
"Bayesian Coalescent Inference of Past Population Dynamics from Molecular Sequences". Molecular Biology and Evolution. 22 (5): ... Shapiro, Beth (2015). How to Clone a Mammoth: The Science of De-Extinction. Princeton, NJ: Princeton University Press. ISBN ... Shapiro's research on ecology has been published in leading journals[3] including Molecular Biology and Evolution,[12] PLOS ... Beth Alison Shapiro (born 1976[5]) is an American evolutionary molecular biologist. She is a Professor in the Department of ...
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. 42. pp. 259-87. doi:10.1016/s0079-6603(08)60578-4. ISBN 9780125400428 ...
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". ...
For their use as vectors, and for molecular cloning, plasmids often need to be isolated. There are several methods to isolate ... A plasmid cloning vector is typically used to clone DNA fragments of up to 15 kbp. To clone longer lengths of DNA, lambda phage ... Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA ... Russell, David W.; Sambrook, Joseph (2001). Molecular cloning: a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor ...
Molecular cloning and functional characterization". The Journal of Biological Chemistry. 277 (26): 23294-300. doi:10.1074/jbc. ... Molecular Cell. 13 (5): 677-88. doi:10.1016/S1097-2765(04)00083-8. PMID 15023338. Fort P, Blangy A (June 2017). "The ...
Molecular cloning and cofactor characterization". The Journal of Biological Chemistry. 273 (27): 16678-85. doi:10.1074/jbc. ... Liu JQ, Odani M, Yasuoka T, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (July 2000). "Gene cloning and overproduction of ...
Neuberger, M. S.; Honjo, T.; Alt, Frederick W. (2004). Molecular biology of B cells. Amsterdam: Elsevier. pp. 189-191. ISBN 0- ... the activation and growth of B cell clones able to secrete antibodies of higher affinity for the antigen. ...
Methods in Molecular Biology, Springer US, pp. 15-25, doi:10.1007/978-1-4939-9865-4_3, ISBN 9781493998654. , PMID 31541435. ... "Map-Based Cloning of the Gene Associated With the Soybean Maturity Locus E3". Genetics. 182 (4): 1251-1262. doi:10.1534/ ... Methods in Molecular Biology, Springer US, 2072, pp. 15-25, doi:10.1007/978-1-4939-9865-4_3, ISBN 9781493998654. , PMID ... QTLs are mapped by identifying which molecular markers (such as SNPs or AFLPs) correlate with an observed trait. This is often ...
"DNA Cloning Using In Vitro Site-Specific Recombination". Genome Res. 10 (11): 1788-95. PMC 310948. . PMID 11076863. doi ... Molecular function. • follicle-stimulating hormone activity. • ربط بروتيني. • فعالية الهرمون. Cellular component. • ...
"Cloning and characterization of MN1, a gene from chromosome 22q11, which is disrupted by a balanced translocation in a ... Molecular function. • molecular function. Cellular component. • cellular component. Biological process. • multicellular ... "A genome annotation-driven approach to cloning the human ORFeome". Genome Biol. 5 (10): R84. doi:10.1186/gb-2004-5-10-r84. PMC ...
1987). "Molecular cloning and sequencing of a human hepatitis delta (delta) virus RNA". Nature. 329 (6137): 343-346. doi: ... "Molecular Phylogenetic Analyses Indicate a Wide and Ancient Radiation of African Hepatitis Delta Virus, Suggesting a ...
Molecular function. • ion channel activity. • benzodiazepine receptor activity. • chloride channel activity. • extracellular ... Yang W, Drewe JA, Lan NC (1996). "Cloning and characterization of the human GABAA receptor alpha 4 subunit: identification of a ...
Molecular function. • acrosin binding. • GO:0001948 protein binding. • carbohydrate binding. • identical protein binding. • ... van Duin M, Polman JE, Verkoelen CC, Bunschoten H, Meyerink JH, Olijve W, Aitken RJ (December 1992). "Cloning and ... Rankin T, Dean J (2000). "The zona pellucida: using molecular genetics to study the mammalian egg coat". Rev. Reprod. 5 (2): ...
"Molecular Biology of the Cell. 6 (5): 509-24. doi:10.1091/mbc.6.5.509. PMC 301212 . PMID 7545030.. ... and functional expression of cDNA clones". Biochemistry. 30 (44): 10640-6. doi:10.1021/bi00108a006. PMID 1657150.. ... American Journal of Respiratory Cell and Molecular Biology. 24 (2): 101-7. doi:10.1165/ajrcmb.24.2.4264. PMID 11159042.. ...
Molecular cloning. N. *Non-coding DNA. *Nuclear DNA. P. *Point mutation. *Promoter (genetics) ...
"Molecular and Cellular Biology. 21 (2): 524-33. doi:10.1128/MCB.21.2.524-533.2001. PMC 86614. PMID 11134340.. ... "Chromosomal localization of seven PAX genes and cloning of a novel family member, PAX-9". Nature Genetics. 3 (4): 292-8. doi: ... "Molecular and Cellular Biology. 13 (10): 6024-35. doi:10.1128/mcb.13.10.6024. PMC 364662. PMID 8413205.. ... Molecular function. • DNA binding. • sequence-specific DNA binding. • transcription factor activity, sequence-specific DNA ...
Molecular Biology and Evolution 21 (1): 14-28. PMID 12949138. doi:10.1093/molbev/msg224. Cite uses deprecated parameter ,month= ... "Identification of duplicated fourth alpha2-adrenergic receptor subtype by cloning and mapping of five receptor genes in ...
"Molecular and Cellular Biology. 20 (9): 3102-15. doi:10.1128/MCB.20.9.3102-3115.2000. PMC 85605. PMID 10757795.. ... "Complete amino acid sequence of the human progesterone receptor deduced from cloned cDNA". Biochemical and Biophysical Research ... "Molecular and Cellular Biology. 19 (2): 1182-9. doi:10.1128/mcb.19.2.1182. PMC 116047. PMID 9891052.. ... "Molecular and Cellular Biology. 18 (8): 4471-87. doi:10.1128/mcb.18.8.4471. PMC 109033. PMID 9671457.. ...
Hideyuki Mannena & Steven S.-L. Li (1999). "Molecular evidence for a clade of turtles". Molecular Phylogenetics and Evolution. ... In some species of squamates, a population of females is able to produce a unisexual diploid clone of the mother. This form of ... Classically, turtles were considered to be related to the primitive anapsid reptiles.[32] Molecular work has usually placed ... Lee, M.S.Y. (2013). "Turtle origins: Insights from phylogenetic retrofitting and molecular scaffolds". Journal of Evolutionary ...
15,0 15,1 Lajtha, A. (2007). Handbook of neurochemistry and molecular neurobiology: Neural protein metabolism and function, 2nd ... Pimenta, A. F., Fischer, I., Levitt, P. (1996). cDNA cloning and structural analysis of the human limbic-system-associated ... George J. Siegel, R. Wayne Albers, Scott T. Brady, Donald L. Price, Basic neurochemistry: molecular, cellular and medical ...
"Methods in Molecular Biology 226.. *↑ Burke, D. T., el al. (1987). "Cloning of Large Segments of Exogenous DNA into Yeast by ... "Journal of Molecular Biology 302 (1): 205-217.. *↑ Roberts, L. (2001). "A History of the Human Genome Project". Science 291 ( ... Molecular Evolution. ISBN 978-0878934805.. *↑ Pop, M. (2004). "Shotgun Sequence Assembly" (PDF). Advances in Computers 60. ISSN ... "Journal of Molecular Biology 98 (3).. *↑ Sanger, F.; et al. (1977). "DNA sequencing with chain-terminating inhibitors". ...
January - The first animal from an extinct species to be recreated by cloning, a Pyrenean Ibex, is born alive, but dies seven ... American scientist who played a central role in the early history of molecular biology (b. 1912). 29 October - Bei Shizhang, ... Gray, Richard; Dobson, Roger (January 31, 2009). "Extinct ibex is resurrected by cloning". The Daily Telegraph. Retrieved 2011- ... President of the International Society for Molecular and Cell Biology and Biotechnology Protocols and Researches (ISMCBBPR). ( ...
Molecular function. • nucleotide binding. • DNA binding. • DNA-dependent ATPase activity. • recombinase activity. • chromatin ... Shinohara A, Ogawa H, Matsuda Y, Ushio N, Ikeo K, Ogawa T (Jul 1993). "Cloning of human, mouse and fission yeast recombination ... "Nature Structural & Molecular Biology. 17 (10): 1247-54. doi:10.1038/nsmb.1915. PMC 4094107 . PMID 20871615.. ... Klopfleisch R, von Euler H, Sarli G, Pinho SS, Gärtner F, Gruber AD (Jan 2011). "Molecular carcinogenesis of canine mammary ...
... and a poliovirus clone was the first infectious DNA clone made of an RNA virus in animals. Along with rhinovirus, poliovirus ... J Gen Virol 85(Pt 5):1145-1151 Mingxiao M, Ming L, Jian C, Song Y, Shude W, Pengfei L (2011) Molecular and biological ... This was the first time that infection virus had been produced from molecular building blocks in the cells. Polyprotein ... "ICTV Report Picornaviridae". Ryu, W.S, 2016, Molecular Virology of Human Pathogenic Viruses, Academic Press, Korea, Page 153- ...
Obesity wars: Molecular progress confronts an expanding epidemic. Cell (Review). 2004, 116 (2): 337-50. PMID 14744442. doi: ... Positional cloning of the mouse obese gene and its human homologue.. Nature (Research Support). 1994-12-01, 372 (6505): 425-32 ... Barness LA, Opitz JM, Gilbert-Barness E. Obesity: genetic, molecular, and environmental aspects. American Journal of Medical ... Boulpaep, Emile L.; Boron, Walter F. Medical physiology: A cellular and molecular approach. Philadelphia: Saunders. 2003: 1227 ...
"Molecular cloning of the human T-lymphocyte surface CD2 (T11) antigen.". Proc. Natl. Acad. Sci. U.S.A. 83 (22): 8718-22. PMC ... "Molecular cloning and expression of T11 cDNAs reveal a receptor-like structure on human T lymphocytes.". Proc. Natl. Acad. Sci ... "Molecular cloning of the CD2 antigen, the T-cell erythrocyte receptor, by a rapid immunoselection procedure.". Proc. Natl. Acad ... Wilkins A, Yang W, Yang J (2003). "Structural biology of the cell adhesion protein CD2: from molecular recognition to protein ...
Kao CY and Levinson SR (1986) Tetrodotoxin, saxitoxin, and the molecular biology of the sodium channel New York Academy of ... Carmichael WW, Gorham PR (1978). "Anatoxins from clones of Anabaena flos-aquae isolated from lakes of western Canada". Mitt. ... Gerald Karp (19 October 2009). Cell and Molecular Biology: Concepts and Experiments. John Wiley and Sons. pp. 14-. ISBN 978-0- ... Herrero A and Flores E (editor). (2008). The Cyanobacteria: Molecular Biology, Genomics and Evolution. Caister Academic Press. ...
Further, to help understand the neurobiology of behavior, fox and dog orthologs of serotonin receptor genes were cloned.[9] ... Trut and her colleagues have applied modern molecular techniques to the fox populations with the aim of not only identifying ...
Molecular function. • calcium channel activity. • metal ion binding. • voltage-gated ion channel activity. • ion channel ... "Cloning, chromosomal localization, and functional expression of the alpha 1 subunit of the L-type voltage-dependent calcium ... "Molecular Psychiatry. 15 (10): 1016-22. doi:10.1038/mp.2009.49. PMC 3011210. PMID 19621016.. ... Perez-Reyes E, Wei XY, Castellano A, Birnbaumer L (Nov 1990). "Molecular diversity of L-type calcium channels. Evidence for ...
... molecular biology by allowing the polymerase chain reaction to be used in research as a simple and rapid technique for cloning ... Cellular and Molecular Life Sciences (CMLS). 54 (4): 305-308. doi:10.1007/s000180050156.. ... Garrett RA; Klenk H (2005). Archaea: Evolution, Physiology and Molecular Biology. WileyBlackwell. ISBN 1-4051-4404-1.. ... Cavicchioli R (2007). Archaea: Molecular and Cellular Biology. American Society for Microbiology. ISBN 1-55581-391-7.. ...
Dearry A, Gingrich JA, Falardeau P, Fremeau RT Jr, Bates MD, Caron MG (1990). "Molecular cloning and expression of the gene for ... Zhou QY, Grandy DK, Thambi L, Kushner JA, Van Tol HH, Cone R, Pribnow D, Salon J, Bunzow JR, Civelli O (1990). "Cloning and ...
A new gene located on chromosome 2 was named timeless (tim) and was successfully cloned and sequenced. They found strong ... "for their discoveries of molecular mechanisms controlling the circadian rhythm".[3][4] ... which led to his future work in cloning the period gene.[6] ... University School of Medicine with an interest in molecular ...
Reddy SV, Zhou ZQ, Rao KJ, et al. (1989). „Molecular characterization of human factor XSan Antonio". Blood 74 (5), 1486-90. o. ... Messier TL, Pittman DD, Long GL, et al. (1991). „Cloning and expression in COS-1 cells of a full-length cDNA encoding human ... Jagadeeswaran P, Reddy SV, Rao KJ, et al. (1990). „Cloning and characterization of the 5' end (exon 1) of the gene encoding ... Cooper DN, Millar DS, Wacey A, et al. (1997). „Inherited factor X deficiency: molecular genetics and pathophysiology". Thromb. ...
"Molecular and Cellular Biology. 7 (1): 541-544. doi:10.1128/MCB.7.1.541. PMC 365100. PMID 3550423.. ... GTPase activity of different mutant forms of p21, one cloned from a patient with myeloblastic leukaemia and one derived from in ... "Molecular and Cellular Biology. 31 (1): 81-91. doi:10.1128/MCB.01001-10. PMC 3019857. PMID 20974804.. ... These findings were published in Molecular and Cellular Biology (MCB).[6] Alan Hall showed the specificity of Rho in the ...
1998). "Molecular cloning and characterization of a human protein kinase that specifically activates c-Jun N-terminal kinase". ...
DNA Peptide biology biosynthesis cloning gene gene expression immune system metabolism physiology protein protein structure ... During the past ten years, refinements in the techniques of recombinant DNA technology have resulted in the cloning of genes ... 1.Laboratory of Molecular Endocrinology and Howard Hughes Medical InstituteMassachusetts General HospitalBostonUSA ... proaches that have been used to identify and select the cloned genes encoding these polypeptides. The contents of the two in- ...
Experts worried about human cloning warn that attempts to clone monkeys -- genetically closer to people -- using the Dolly ... A new study reveals why cloned human and other primate cells dont result in a healthy pregnancy. ... Molecular Block to Viable Clones. WASHINGTON - Cloning humans, or any other primates, may be impossible with todays techniques ... Cloning experts worry that attempting human cloning is dangerous, not just because of all the barnyard clones with birth ...
Molecular Cloning: A Laboratory Manual, Book 1. Molecular Cloning: A Laboratory Manual, Joseph Sambrook, ISBN 0879693096, ... Molecular cloning: a laboratory manual. Vol. 3, Volume 2. Joseph Sambrook,David W. Russell. No preview available - 2001. ... Cloning.html?id=qbATAQAAIAAJ&utm_source=gb-gplus-shareMolecular Cloning. ... Molecular Cloning: 1. Joseph Sambrook,Edward F. Fritsch,Thomas Maniatis. No preview available - 2012. ...
Molecular Cloning: A Laboratory Manual, Book 1. Molecular Cloning: A Laboratory Manual, Joseph Sambrook, ISBN 0879693096, ... Cloning.html?id=qbATAQAAIAAJ&utm_source=gb-gplus-shareMolecular Cloning. ... Molecular Cloning: A Laboratory Manual, Book 1. Joseph Sambrook,Tom Maniatis. Snippet view - 1989. ... Molecular Cloning: A Laboratory Manual, Book 1. Joseph Sambrook,Tom Maniatis. Snippet view - 1989. ...
Molecular Cloning (Process by which gene products are made) PCR product cloning vector ligation transformation expression E. ... the piece of DNA next to the gene must be turned on • By cloning the promoter beta-lactoglobulin and attaching it to the human ... The objective of Genetic engineering in this case is to clone the insulin gene from the human genome. ... After sufficient replication.The gene when extracted from the human genome is ligated unto the cloning vector. bacteriophage ...
Kurogi K., Sakakibara Y., Yasuda S., Liu MC., Suiko M. (2010) Molecular Cloning and Characterization of a Novel Canine ... Tsoi, C., Falany, C.N., Morgenstern, R., and Swedmark, S. (2001) Molecular cloning, expression, and characterization of a ... Tsoi, C., Morgenstern, R., and Swedmark, S. (2002) Canine sulfotransferase SULT1A1: Molecular cloning, expression, and ... Molecular cloning, expression, and characterization of novel human SULT1C sulfotransferases that catalyze the sulfonation of N- ...
Molecular Biology Reports. Methylglyoxal is a kind of poisonous metabolite that can react with RNA, DNA and protein, which ... Molecular cloning and characterization of a novel glyoxalase I gene TaGly I in wheat (Triticum aestivum L.). *Lin F ... The corresponding full length gene, named TaGly I, was cloned, sequenced and characterized. Its genomic sequence consists of ...
... Eur J Biochem. 1996 Dec 15;242(3):822-31. doi: 10.1111/j.1432-1033.1996. ...
The development of gene cloning vectors and selection methodologies enabled the cloning of REases. Cloning not only allowed the ... Restriction Endonucleases: Molecular Cloning and Beyond. Return to Restriction Endonucleases The sequence-specific DNA cleavage ... Home Products Restriction Endonuclease Products Restriction Endonucleases Restriction Endonucleases: Molecular Cloning and ... Restriction enzymes have been one of the major forces that enabled the cloning of genes and transformed molecular biology. ...
Molecular Cloning and Beyond Restriction Endonucleases: Molecular Cloning and Beyond. The sequence-specific DNA cleavage ... The development of gene cloning vectors and selection methodologies enabled the cloning of REases. Cloning not only allowed the ... Restriction enzymes have been one of the major forces that enabled the cloning of genes and transformed molecular biology. ... Innovative applications of these enzymes will take REases role beyond molecular cloning by continuing to accelerate the ...
Accordingly, we undertook molecular cloning and sequence analysis of human endothelial NO synthase. Complementary DNA clones ... Molecular cloning and characterization of human endothelial nitric oxide synthase.. Marsden PA1, Schappert KT, Chen HS, Flowers ... However, recent cloning and molecular characterization of NO synthase from bovine endothelial cells indicated the existence of ...
Molecular cloning, sequence, expression, and processing of the interleukin 16 precursor. Michael Baier, Norbert Bannert, ... Molecular cloning, sequence, expression, and processing of the interleukin 16 precursor. Michael Baier, Norbert Bannert, ... Molecular cloning, sequence, expression, and processing of the interleukin 16 precursor Message Subject (Your Name) has sent ... Molecular cloning, sequence, expression, and processing of the interleukin 16 precursor. Michael Baier, Norbert Bannert, ...
The Condensed Protocols From Molecular Cloning: A Laboratory Manual is a single-volume adaptation of the three-volume third ... Step By Step Laboratory Manual for Condensed Protocols From Molecular Cloning. by Sam Savage ... "Condensed Protocols from Molecular Cloning: A Laboratory Manual" report to their offering. ... edition of Molecular Cloning: A Laboratory Manual. This condensed book contains only the step-by-step portions of the protocols ...
Any techniques related to molecular cloning including restriction digestion, ligation, transformation, plasmid... ... Molecular Cloning. Any techniques related to molecular cloning including restriction digestion, ligation, transformation, ... Designing Crispr guides and cloning into appropriate vectors Started by Natalia KM, 13 Sep 2019 molecular cloning, Crispr/Cas9 ... Cloning of an unknown gene Started by bio_kid, 13 Apr 2015 cloning, genome library and 1 more... * 4 replies ...
Top : Forum Archives: : Molecular Cloning. cloning - (Jan/14/2008 ). I am trying to PCR the digested and dephosphorylated ...
Zheng, T., Rabach, M., Chen, N.Y., Rabach, L., Hu, X., Elias, J.A. and Zhu, Z. (2005) Molecular Cloning and Functional ... Suzuki, T., Kakizaki, H., Ikeda, M. and Matsumiya, M. (2014) Molecular Cloning of a Novel Chitinase Gene from Blue Shark ( ... Kurokawa, T., Tuji, S. and Suzuki, T. (2004) Molecular Cloning of Multiple Chitinase Genes in Japanese Flounder, Paralichthys ... Molecular Cloning, Distribution, and Phylogenetic Analysis. Open Journal of Marine Science, 8, 136-151. doi: 10.4236/ojms. ...
This chapter of the Protocols and Applications Guide provides a background on basic cloning with a focus on cloning PCR ... Background of basic cloning with a focus on cloning PCR fragments Introduction The cloning of genes, gene fragments and other ... T-Cloning Vectors T vectors are a specific type of cloning vector that get their name from the T overhangs added to a ... Promega Products for Cloning Thermostable DNA Polymerases The use of amplification enzymes is the first step in cloning by PCR ...
ATCC offers a growing selection of genomic and cDNA clones to enhance your research. ...
Molecular cloning is a set of methods, which are used to insert recombinant DNA into a vector - a carrier of DNA molecules that ... Critical aspects of molecular cloning are discussed, such as the need for molecular cloning strategy and how to keep track of ... Youve just watched JoVEs video on molecular cloning. You should now understand how molecular cloning works and how the ... Molecular Cloning. JoVE, Cambridge, MA, (2018).. Molecular cloning is a set of techniques used to insert recombinant DNA from a ...
Molecular cloning of a novel cytoplasmic protein tyrosine phosphatase PTP epsilon.. Nakamura K1, Mizuno Y, Kikuchi K. ...
Human acetyl-CoA carboxylase: characterization, molecular cloning, and evidence for two isoforms.. L Abu-Elheiga, A Jayakumar, ... Human acetyl-CoA carboxylase: characterization, molecular cloning, and evidence for two isoforms. ... Human acetyl-CoA carboxylase: characterization, molecular cloning, and evidence for two isoforms. ... Human acetyl-CoA carboxylase: characterization, molecular cloning, and evidence for two isoforms. ...
Other articles related to molecular cloning, molecular, cloning:. Clone (genetics) - Overview. ... Molecular cloning takes ... Read more about Molecular Cloning: History of Molecular Cloning, Overview, Steps in Molecular Cloning, Applications of ... Molecular Cloning. Molecular cloning is a set of experimental methods in molecular biology that are used to assemble ... Molecular Cloning. ... Molecular cloning refers to the process of making multiple molecules ... Cloning is commonly used to ...
... Mol Cell Neurosci. 1990 Dec;1(3):214-23. doi: ...
Molecular Cloning. by Sambrook, J., Fritsch, E. F., Maniatis, T. by Sambrook, J., Fritsch, E. F., Maniatis, T. Recommend this! ... Sambrook, J. is the author of Molecular Cloning, published 1989 under ISBN 9780879693732 and ISBN 0879693738. ...
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 ...
... Seow-Neng Chan,1 Norliza Abu ... Molecular Cloning of CypCl Gene and Sequence Analysis. Once the PCR product was proven to show similarity to other ... P. Bangrak and W. Chotigeat, "Molecular cloning and biochemical characterization of a novel cystatin from Hevea rubber latex," ... W. Megdiche, C. Passaquet, W. Zourrig, Y. Zuily Fodil, and C. Abdelly, "Molecular cloning and characterization of novel ...
Topics cover isolation of gene fragment, screening, polymerase chain reaction, cloning vectors, and the isolation of plasmids. ... This free online cloning course provides a comprehensive knowledge of the fundamental processes involved in molecular cloning. ... Introduction to Molecular Cloning This free online course on Alison takes you through the concepts, processes, and significance ... Introduction to Molecular Cloning is a free online course that begins by discussing the procedure for isolating specific genes ...
Get more protocols at molecular cloning central ». Want to discover the easiest way to clone?. The easiest way to clone is to ... Struggling to clone your gene of interest or wasting precious time troubleshooting molecular cloning procedures? Our Molecular ... What are the advantages of GenEZ™ cDNA ORF Clones?. *Convenient: cDNA ORF clones available in the vector of your choice (choose ... Start by searching for your ORF sequence in our ORF clone database below, or if you know the accession number of your clone, ...
Molecular cloning and characterization of an insulin-regulatable glucose transporter [29].. *Molecular cloning of cDNAs ... Disease relevance of Cloning, Molecular. *Amplification and Molecular Cloning of HTLV-I Sequences from DNA of Multiple ... High impact information on Cloning, Molecular. *Molecular cloning of NCR revealed novel members of the Ig superfamily ... Biological context of Cloning, Molecular. *From peptide sequence, molecular cloning revealed a cDNA encoding a novel protein, ...
Molecular cloning and duplication of the nematode sex-determining gene tra-1.. J Hodgkin ... Molecular cloning and duplication of the nematode sex-determining gene tra-1.. J Hodgkin ... Molecular cloning and duplication of the nematode sex-determining gene tra-1.. J Hodgkin ... Molecular cloning and duplication of the nematode sex-determining gene tra-1. ...
  • Falany, C. and Roth, J.A. (1993) In: Jeffery, E.H. (Ed.), Human Drug Metabolism: From Molecular Biology to Man (pp. 101-115). (
  • The sequence-specific DNA cleavage activity of restriction endonucleases (REases), combined with other enzymatic activities that amplify and ligate nucleic acids, have enabled modern molecular biology. (
  • Our investigation of the molecular biology of IL-16 has therefore subsequently focused on the characterization of this initially elusive IL-16 precursor. (
  • This condensed book contains only the step-by-step portions of the protocols, accompanied by selected appendices from the world's best-selling manual of molecular biology techniques. (
  • 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. (
  • Molecular cloning methods are central to many contemporary areas of modern biology and medicine. (
  • Prior to the 1970s, the understanding of genetics and molecular biology was severely hampered by an inability to isolate and study individual genes from complex organisms. (
  • MRC Laboratory of Molecular Biology, Cambridge, England. (
  • A vector is plasmid DNA used as a tool in molecular biology to make more copies of or produce a protein from a certain gene. (
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  • Since then, molecular cloning has become one of the most powerful tools of the molecular biology laboratory enabling the expression of the smallest genes, as well as the engineering of whole genomes. (
  • Dublin, Aug. 21, 2014 (GLOBE NEWSWIRE) -- Research and Markets ( ) has announced the addition of the "Global Molecular Biology Enzymes, Kits, & Reagents (Cloning, Epigenetics, PCR, Restriction Digestion, Sequencing) Market - Forecast to 2018" report to their offering. (
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  • For the fourth edition of this classic work, the content has been entirely recast to include nucleic-acid based methods selected as the most widely used and valuable in molecular and cellular biology laboratories. (
  • Building on thirty years of trust, reliability, and authority, the fourth edition of Molecular Cloning is the new gold standard the one indispensable molecular biology laboratory manual and reference source. (
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  • cDNA cloning and gene expression of cecropin D, an antibacterial protein in the silkworm, Bombyx mori," Comparative Biochemistry and Physiology-B Biochemistry and Molecular Biology , vol. 122, no. 4, pp. 409-414, 1999. (
  • Characterization of a Bombyx mori cDNA encoding a novel member of the attacin family of insect antibacterial proteins," Insect Biochemistry and Molecular Biology , vol. 25, no. 3, pp. 385-392, 1995. (
  • Institute of Molecular Biology I, University of Zurich, Switzerland. (
  • The first two editions of this manual have been mainstays of molecular biology for nearly twenty years, with an unrivalled reputation for reliability, accuracy, and clarity. (
  • In this new edition, authors Joe Sambrook and David Russell have completely updated the book, revising every protocol and adding a mass of new material, to broaden its scope and maintain its unbeatable value for studies in genetics, molecular cell biology, developmental biology, microbiology, neuroscience, and immunology. (
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  • Message Body (Your Name) thought you would be interested in this article in Molecular and Cellular Biology. (
  • In this volume, top scientists in these fields provide perspectives on how molecular data in biology help to elucidate key questions in estimating paleontological divergence and in understanding the mechanisms behind phenotypic evolution. (
  • Paleobiological questions such as genome size, digit homologies, genetic control cascades behind phenotype, estimates of vertebrate divergence dates, and rates of morphological evolution are addressed, with a special emphasis on how molecular biology can inform paleontology, directly and indirectly, to better understand life's past. (
  • Highlighting a significant shift towards interdisciplinary collaboration, this is a valuable resource for students and researchers interested in the integration of organismal and molecular biology. (
  • Following the cloning of NaPi-IIa [ 33 ], the phosphate physiology field has progressed rapidly, benefitting from advances in molecular and cell biology, imaging, and biophysical assays, as exemplified by the invited articles. (
  • Working at the forefront of molecular biology, Maniatis has perfected protocols for identifying and isolating genes and has used this methodology to decipher how different genes are activated in response to the environment. (
  • Soon after, the discovery and purification of REases that recognized and cut at specific sites (Type II REases) allowed scientists to perform precise manipulations of DNA in vitro , such as the cloning of exogenous genes and creation of efficient cloning vectors. (
  • The development of gene cloning vectors and selection methodologies enabled the cloning of REases. (
  • That is, these plasmids could serve as cloning vectors to carry genes. (
  • By the end of the course, you will be familiar with the roles and importance of cloning vectors. (
  • Finally, you will be introduced to the transforming agents known as cloning vectors and the determinants of a good vector. (
  • Molecular cloning is a set of techniques used to insert recombinant DNA from a prokaryotic or eukaryotic source into a replicating vehicle such as plasmids or viral vectors. (
  • E. coli grows rapidly, is widely available and has numerous different cloning vectors commercially produced. (
  • A MCS is found in a variety of vectors, including cloning vectors to increase the number of copies of target DNA, and in expression vectors to create a protein product. (
  • During the past ten years, refinements in the techniques of recombinant DNA technology have resulted in the cloning of genes encoding approximately 50 different hormonal and regulatory peptides, including those in which the peptides themselves and the mRNAs encoding the peptides are present in only trace amounts in the tissues of origin. (
  • Two of the objectives in the assembly of this book are to pre- sent, in one volume, the known primary structures of the genes encoding several of the polypeptide hormones and related regulatory peptides, and to provide an account of the various ap- proaches that have been used to identify and select the cloned genes encoding these polypeptides. (
  • The contents of the two in- troductory chapters are intended to provide the reader with a brief background of the approaches to gene cloning and the struc- ture and expression of hormone-encoding genes. (
  • Introduction to Molecular Cloning is a free online course that begins by discussing the procedure for isolating specific genes. (
  • He then became excited by DNA and worked on mitochondrial genes in fungi in order to learn the new (in those days) techniques for gene cloning and DNA sequencing. (
  • To define the HA mutations related to these antigenic sites, we cloned and sequenced the HA genes amplified by polymerase chain reaction of parent virus and MAb-selected escape mutants. (
  • Molecular cloning and characterization of annexin genes in peanut (Arachis hypogaea L. (
  • Cloning protocols for using the pLKO.1 vector, a backbone used by the RNAi consortium for targeting human and mouse genes. (
  • Molecular cloning and characterization of acrA and acrE genes of Escherichia coli. (
  • The concluding chapters deal with methods to screen expression libraries, express cloned genes in both prokaryotes and eukaryotic cells, analyze transcripts and proteins, and detect protein-protein interactions. (
  • Among these, causative genes for inherited diseases of the retina have been identified by positional cloning or candidate gene approaches. (
  • Thus, isolating genes uniquely expressed in the retina could lead to the clarification of the molecular mechanisms of phototransduction or the development of retinal disorders. (
  • Efstratiadis was interested in establishing methods for cloning mammalian genes. (
  • Multiple cloning sites are a feature that allows for the insertion of foreign DNA without disrupting the rest of the plasmid which makes it extremely useful in biotechnology, bioengineering, and molecular genetics. (
  • Research and Markets ( has announced the addition of the "Condensed Protocols from Molecular Cloning: A Laboratory Manual" report to their offering. (
  • The Condensed Protocols From Molecular Cloning: A Laboratory Manual is a single-volume adaptation of the three-volume third edition of Molecular Cloning: A Laboratory Manual. (
  • For corrections to Molecular Cloning: A Laboratory Manual, Fourth Edition, click HERE . (
  • Molecular Cloning: A Laboratory Manual fills the same niche in the laboratory (with) information to help both the inexperienced and the advanced user. (
  • It) has once again established its primacy as the molecular laboratory manual and is likely to be found on lab benches. (
  • Molecular Cloning: A Laboratory Manual has always been the laboratory mainstay for protocols and techniques. (
  • the next generation of Molecular Cloning not only carries on the proud heritage of the first two editions but also admirably expands on that tradition to provide a truly essential laboratory manual. (
  • Molecular Cloning: A Laboratory Manual came out in 1982. (
  • Complementary DNA clones predict a protein of 1,203 amino acids sharing 94% identity with the bovine endothelial protein, but only 60% identity with the rat brain NO synthase isoform. (
  • In earlier investigations the good agreement between the apparent molecular mass of IL-16 in SDS/PAGE (17 kDa) and an open reading frame of 390 bp suggested that IL-16 is produced as a mature 130-amino acid protein with neither a signal peptide nor further processing ( 10 ). (
  • However, we recently found that the originally cloned IL-16 ( 10 ) is part of a much longer open reading frame ORF, indicating that the 17 kDa protein is derived from a corresponding precursor ( 14 ). (
  • Molecular cloning of a novel cytoplasmic protein tyrosine phosphatase PTP epsilon. (
  • The cDNA isolated from a clone library consisted of 2772 bp containing a 2115 bp open reading frame coding for a 705 aa protein with an approximate molecular mass of 80 kDa. (
  • Internal amino acid sequence was obtained from this 120,000-dalton protein and, by means of this sequence, two types of complementary DNA clones were isolated and characterized. (
  • The recombinant protein showed an estimated molecular mass of 87 kDa, consistent with the expected value of the annotated protein, and an optimal activity for the hydrolysis of α-PNPG was detected at 40 °C, and at pH 6.0. (
  • Molecular cloning and expression study of Xenopus latent TGF-beta binding protein-1 (LTBP-1). (
  • Sequence analysis of XLTBP-1 cDNA revealed an open reading frame of 4518 bp encoding a 1398 amino acid protein with a molecular mass of 154.1 kDa and an isoelectric point of 4.65. (
  • deglycosylation studies indicate that four of the five N-linked glycosylation sites are used and that the mature core protein has a molecular mass of 25 kDa, close to that predicted by the cDNA sequence. (
  • Zebrafish P-selectin cDNA is 2,800 bp and encodes a putative 868 amino acid protein with a theoretical molecular weight of 122.36 kDa and isoelectric point of 6.27. (
  • Sequence analysis indicated that the cloned PIPK gene had a high degree of homology with other mammalian PIPKs at the DNA and protein level. (
  • In this study, a cDNA encoding a novel canine SULT was cloned from MDCK cells using reverse transcription-polymerase chain reaction (RT-PCR) technique. (
  • With the advent of the Polymerase Chain Reaction (PCR), RT-PCR, and PCR-based mutagenesis methodologies, the traditional cloning workflow transformed biological research in the decades that followed. (
  • Cloning and characterization of a Xenopus poly(A) polymerase. (
  • In this study, we cloned the full-length cDNA of P-selectin from zebrafish ( Danio rerio ) by the method of rapid amplification of cDNA ends polymerase chain reaction (RACE-PCR). (
  • Molecular cloning is a set of methods, which are used to insert recombinant DNA into a vector - a carrier of DNA molecules that will replicate recombinant DNA fragments in 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 . (
  • Strictly speaking, recombinant DNA refers to DNA molecules, while molecular cloning refers to the experimental methods used to assemble them. (
  • 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. (
  • Microbiologists, seeking to understand the molecular mechanisms through which bacteria restricted the growth of bacteriophage, isolated restriction endonucleases, enzymes that could cleave DNA molecules only when specific DNA sequences were encountered. (
  • At least two important DNA molecules are required before cloning begins. (
  • The opening chapters describe essential techniques, some well-established, some new, that are used every day in the best laboratories for isolating, analyzing and cloning DNA molecules, both large and small. (
  • We have isolated a cDNA clone (mERD2) for the mammalian (bovine) homologue of the yeast ERD2 gene, which codes for the yeast HDEL receptor. (
  • The cDNA clone was ligated into a mammalian expression vector and transfected into COS-1 cells. (
  • One bacterial plasmid used in genetic engineering as a plasmid cloning vector is pUC18. (
  • Learn about genetic engineering, gene manipulation and modification acheived by molecular cloning and transformation. (
  • The kits and reagents product segment has five sub-segments, namely, cloning and mutagenesis, nucleic acid analysis, PCR sequencing, and other kits and reagents. (
  • These are followed by chapters on cDNA cloning and exon trapping, amplification of DNA, generation and use of nucleic acid probes, mutagenesis, and DNA sequencing. (
  • The DNA fragment containing the acrA locus of the Escherichia coli chromosome has been cloned by using a complementation test. (
  • Molecular cloning and characterization of human endothelial nitric oxide synthase. (
  • We have cloned, sequenced, and examined several biochemical properties of a Xenopus PAP. (
  • Molecular cloning, characterization, and chromosomal mapping of a novel human gene ( GTF3A ) that is highly homologous to Xenopus transcription factor IIIA. (
  • Using molecular cloning techniques, human homologs of the known members of the trk family of neurotrophin receptors have been cloned and sequenced. (
  • Molecular cloning and characterization of crustacean type-one dopamine receptors: D1alphaPan and D1betaPan. (
  • As a first step toward illuminating the molecular underpinnings of dopaminergic signal transduction in the crustacean STNS, we have cloned and characterized two type-one DA receptors (DARs) from the spiny lobster (Panulirus interruptus): D(1alphaPan) and D(1betaPan). (
  • Transmembrane proteins corresponding to the ligand binding chains of these receptors have recently been cloned. (
  • The cDNA cloning was based on the structural homology to hepatic asialoglycoprotein receptors. (
  • 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. (
  • In this study, the full-length cDNA of pig Cd127 was cloned and sequenced. (
  • In the present study, we cloned the full-length cDNA of zebrafish P-selectin and analyzed its expression pattern. (
  • Fujimori, K. , Fukushima, H. and Matsumiya, M. (2018) Molecular Cloning and Phylogenetic Analysis of a Chitin Deacetylase Isolated from the Epidermis of the Red Snow Crab Chionoecetes japonicas . (
  • Molecular cloning and functional characterization of a new Cap'n' collar family transcription factor Nrf3. (
  • Nucleotide sequence revealed that the newly cloned canine SULT cDNA has an open reading frame encompassing 912 nucleotides and encoding a 303-amino acid polypeptide. (
  • Cloning, nucleotide sequence analysis, and characterization of cDNA for medaka (Oryzias latipes) transferrin. (
  • The nucleotide sequence of this cDNA clone was homologous to those of the galactose- and N-acetylgalactosamine-specific C-type macrophage lectins of rodents. (
  • Accordingly, we undertook molecular cloning and sequence analysis of human endothelial NO synthase. (
  • Sequence analysis demonstrated varying homologies with TnC complementary DNA clones isolated from developing chick skeletal muscle. (
  • One type encoded GAP with a predicted molecular mass of 116,000 daltons and 96% identity with bovine GAP. (
  • The second type appeared to be generated by a differential splicing mechanism and encoded a novel form of GAP with a predicted molecular mass of 100,400 daltons. (
  • The 855 bp cDNA of this gene contains the open reading frame (ORF) encoding 232 amino acids with a predicted molecular mass of approx. (
  • To investigate the uptake mechanism, the novel Oatp1d1 in rainbow trout (rtOatp1d1) was cloned, identified and characterized. (
  • A new Oatp1d1 in rainbow trout (rtOatp1d1) was cloned, identified and characterized. (
  • This affordable companion volume, designed for bench use, offers individual investigators the opportunity to have their own personal collection of short protocols from the essential Molecular Cloning. (
  • Welcome to cloning central - a centralized portal for molecular cloning techniques, protocols, and troubleshooting guides . (
  • Core chapters from the third edition have been revised to feature current strategies and approaches to the preparation and cloning of nucleic acids, gene transfer, and expression analysis. (
  • Molecular cloning, expression analysis and cellular localization of an LFRFamide gene in the cuttlefish Sepiella japonica. (
  • Molecular cloning, characterization and expression analysis of S- adenosyl- L-homocysteine hydrolase (SAHH) during the pathogenic infection of Litopenaeus vannamei by Vibrio alginolyticus. (
  • 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 . (
  • Therefore, if any segment of DNA from any organism is inserted into a DNA segment containing the molecular sequences required for DNA replication, and the resulting recombinant DNA is introduced into the organism from which the replication sequences were obtained, then the foreign DNA will be replicated along with the host cell's DNA in the transgenic organism. (
  • In standard molecular cloning experiments, the cloning of any DNA fragment essentially involves seven steps: (1) Choice of host organism and cloning vector, (2) Preparation of vector DNA, (3) Preparation of DNA to be cloned, (4) Creation of recombinant DNA, (5) Introduction of recombinant DNA into host organism, (6) Selection of organisms containing recombinant DNA, (7) Screening for clones with desired DNA inserts and biological properties. (
  • Now, human complementary DNA clones corresponding to three polypeptides present in this BMP preparation have been isolated, and expression of the recombinant human proteins have been obtained. (
  • High accuracy robotics combined with gentle fluidics-based systems establish viable clones with much higher efficiency. (
  • A doctor who separately is pursuing human cloning has reported in an Internet journal preliminary data on an early-stage cloned human embryo, but with no chromosome information. (
  • Indeed, inside cloned monkey cells, the Pittsburgh researchers discovered deformed spindles and chaotic chromosome numbers. (
  • involving the use of restriction enzymes DNA sequencing (where the sequence of nucleotides is known and the gene can be synthesized in the laboratory) Nucleic acid hybridization (used to locate the target gene on its chromosome) DNA cloning (the gene is inserted into the genome of a foreign organism (host) and is replicated as the cells in the host divide). (
  • We have isolated a full length complementary DNA clone (pCTnC1) from a 19-day embryonic chicken heart library corresponding to cardiac troponin C (TnC). (
  • Quarto N (2002), Molecular cloning and expression study of Xenop. (
  • From the very first step, cloned primate cells don't divide properly, causing a helter-skelter mix of chromosomes too abnormal for pregnancy to even begin, University of Pittsburgh researchers reported Thursday in the journal Science. (
  • It also means the related field of therapeutic cloning - using embryonic stem cells to grow customized tissues for medical treatment - may prove harder, too, Schatten said. (
  • Schatten then pulls out that sperm and egg DNA, leaving just the clone DNA in the now-growing monkey cells. (
  • However, recent cloning and molecular characterization of NO synthase from bovine endothelial cells indicated the existence of a family of constitutive NO synthases. (
  • The complete pro-IL-16 cDNA was subsequently molecularly cloned, sequenced, and expressed in COS-7 cells. (
  • The fundamental difference between the two methods is that molecular cloning involves replication of the DNA in a living microorganism, while PCR replicates DNA in an in vitro solution, free of living cells. (
  • George S, Leaver M, Frerichs GN & Burgess D (1989) Fish metallothioneins: Molecular cloning studies and induction in cultured cells, Marine Environmental Research , 28 (1-4), pp. 173-177. (
  • RNA was isolated from Z210R.1 cells, oligo-dT-primed cDNA was synthesized and cloned into the pDC406 vector, pools of clones were generated, and transfections into CV-1/EBNA cells were performed, all as described ( 8 ). (
  • Examples of the DNA sequences that are difficult to clone are inverted repeats, origins of replication, centromeres and telomeres. (
  • This deficit is largely or wholly due to the presence of sequences that cannot be cloned in rec+bacterial hosts. (
  • Start by searching for your ORF sequence in our ORF clone database below, or if you know the accession number of your clone, place a quote directly by clicking the button at the right. (
  • In vivo methyl labeling experiments with L-[methyl-3H]methionine showed that this knockout mutant lacked an MCP with a molecular weight of approximately 68,000. (
  • Brain circuit assemblies comprise different cellular subpopulations that exhibit morphological, electrophysiological, and molecular diversity. (
  • This study will prospectively characterize the molecular, cellular and genetic properties of primary and metastatic neuroblastoma, osteosarcoma, retinoblastoma, Ewing sarcoma family of tum. (
  • The GTP cyclohydrolase I (GTP-CH) gene of the cellular slime mould Dictyostelium discoideum has been cloned and sequenced. (
  • The easiest way to clone is to let GenScript do the work for you through our sequence-verified, expression-ready, GenEZ™ cDNA ORF clones and ORF mutant clones . (
  • What are the advantages of GenEZ™ cDNA ORF Clones? (
  • Known world-wide as the standard introductory text to this important and exciting area, the seventh edition of Gene Cloning and DNA Analysis addresses new and growing areas of research whilst retaining the philosophy of the previous editions. (
  • Professor Brown has written a number of undergraduate textbooks including Gene Cloning and DNA Analysis: An Introduction (6th edition, Wiley-Blackwell, 2010) and Genomes (3rd edition, Garland Science, 2006). (
  • Pellet P, Berger R, Bernheim A, Brouet J, Tsapis A. Molecular analysis of a t(9;14)(p11;q32) translocation occurring in a case of human alpha heavy chain disease. (
  • I. A multivariate analysis of the binding profiles of 14 drugs at 21 native and cloned human receptor subtypes. (
  • Molecular cloning and functional analysis of a blue light receptor gene MdCRY2 from apple (Malus domestica). (
  • Moirangthem LD, Ibrahim KS, Vanlalsangi R, Stensjö K, Lindblad P, Bhattacharya J. Molecular Cloning and Biochemical Characterization of the Iron Superoxide Dismutase from the Cyanobacterium Nostoc punctiforme ATCC 29133 and Its Response to Methyl Viologen-Induced Oxidative Stress. (
  • Borel F, Mueller C. Design, Cloning, and In Vitro Screening of Artificial miRNAs to Silence Alpha-1 Antitrypsin. (
  • This video explains the major methods that are combined, in tandem, to comprise the overall molecular cloning procedure. (
  • In this video you will learn about the different steps of molecular cloning, how to set up the procedure, and different applications of this technique. (
  • The first step of the general molecular cloning procedure is to obtain the desired insert, which can be derived from DNA or mRNA from any cell type. (
  • This can be advantageous when an insert contains a number of restriction sites within its sequence, making it difficult to identify restriction enzymes that will not cut the gene of interest during the cloning procedure. (
  • I am a master student and I am doing my master thesis in the cloning of keratinase sequence in competent cell.I tried several different methods and procedure for the cloning and I got the colonies but when I did the mini prep I had the good concentration of the DNA but when run on agarosei could not see the band. (
  • This changed dramatically with the advent of molecular cloning methods. (
  • A) The sequence given is a composite of three clones (see Materials and Methods). (
  • If you are a Scientist with extensive lentiviral vector design/production, vector construction, and DNA cloning experience, please read on! (
  • Cloning experts worry that attempting human cloning is dangerous , not just because of all the barnyard clones with birth defects, but because attempts to clone monkeys - far closer genetically to people - using the Dolly technique so far have failed. (
  • The objective of Genetic engineering in this case is to clone the insulin gene from the human genome. (
  • The gene when extracted from the human genome is ligated unto the cloning vector. (
  • Human acetyl-CoA carboxylase: characterization, molecular cloning, and evidence for two isoforms. (
  • You'll also find discussions on the health of genetically modified foods and the ethics behind cloning and human genetic engineering. (
  • The murine clone has 61% nucleotide identity with the human cDNA. (
  • Molecular cloning and expression of cDNA encoding human macrophage C-type lectin. (
  • A human macrophage calcium-dependent (C-type) lectin cDNA clone was obtained from a library derived from IL-2-treated peripheral blood monocytes. (
  • Eggs harbor proteins that act as molecular motors that are key to spindle formation. (
  • The insert from this clone, clone 17, was then used to screen an oligo-dT-primed Z210R.1 cDNA library made in λgt10, from which two other clones (2-1 and 1A) were isolated. (
  • Although there is no stretch of polyA at the 3′ end of our clones, it is likely that the AATAAA sequence at nucleotides 1111-1116 represents a polyadenylation signal, since the clone containing it was isolated from an oligo-dT-primed cDNA library. (
  • This method can also be used to add new restriction sites to a multiple cloning site. (
  • A cDNA for plaice metallothionein (MT) was cloned and the sequence for the first 45 amino acid residues was deduced. (
  • XlGSTM1-1 is composed of 219 amino acid residues with a calculated molecular mass of 25359Da. (