Protein analogs and derivatives of the Aequorea victoria green fluorescent protein that emit light (FLUORESCENCE) when excited with ULTRAVIOLET RAYS. They are used in REPORTER GENES in doing GENETIC TECHNIQUES. Numerous mutants have been made to emit other colors or be sensitive to pH.
Proteins which are involved in the phenomenon of light emission in living systems. Included are the "enzymatic" and "non-enzymatic" types of system with or without the presence of oxygen or co-factors.
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.
Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye.
Genes whose expression is easily detectable and therefore used to study promoter activity at many positions in a target genome. In recombinant DNA technology, these genes may be attached to a promoter region of interest.
The class of true jellyfish, in the phylum CNIDARIA. They are mostly free-swimming marine organisms that go through five stages in their life cycle and exhibit two body forms: polyp and medusa.
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 property of emitting radiation while being irradiated. The radiation emitted is usually of longer wavelength than that incident or absorbed, e.g., a substance can be irradiated with invisible radiation and emit visible light. X-ray fluorescence is used in diagnosis.
A light microscopic technique in which only a small spot is illuminated and observed at a time. An image is constructed through point-by-point scanning of the field in this manner. Light sources may be conventional or laser, and fluorescence or transmitted observations are possible.
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 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.
Compound such as LUMINESCENT PROTEINS that cause or emit light (PHYSICAL LUMINESCENCE).
Genes that are introduced into an organism using GENE TRANSFER TECHNIQUES.
Laboratory mice that have been produced from a genetically manipulated EGG or EMBRYO, MAMMALIAN.
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 process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport.
Established cell cultures that have the potential to propagate indefinitely.
The introduction of functional (usually cloned) GENES into cells. A variety of techniques and naturally occurring processes are used for the gene transfer such as cell hybridization, LIPOSOMES or microcell-mediated gene transfer, ELECTROPORATION, chromosome-mediated gene transfer, TRANSFECTION, and GENETIC TRANSDUCTION. Gene transfer may result in genetically transformed cells and individual organisms.
ANIMALS whose GENOME has been altered by GENETIC ENGINEERING, or their offspring.
Substances used for the detection, identification, analysis, etc. of chemical, biological, or pathologic processes or conditions. Indicators are substances that change in physical appearance, e.g., color, at or approaching the endpoint of a chemical titration, e.g., on the passage between acidity and alkalinity. Reagents are substances used for the detection or determination of another substance by chemical or microscopical means, especially analysis. Types of reagents are precipitants, solvents, oxidizers, reducers, fluxes, and colorimetric reagents. (From Grant & Hackh's Chemical Dictionary, 5th ed, p301, p499)
Light-induced change in a chromophore, resulting in the loss of its absorption of light of a particular wave length. The photon energy causes a conformational change in the photoreceptor proteins affecting PHOTOTRANSDUCTION. This occurs naturally in the retina (ADAPTATION, OCULAR) on long exposure to bright light. Photobleaching presents problems when occurring in PHOTODYNAMIC THERAPY, and in FLUORESCENCE MICROSCOPY. On the other hand, this phenomenon is exploited in the technique, FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING, allowing measurement of the movements of proteins and LIPIDS in the CELL MEMBRANE.
The transfer of bacterial DNA by phages from an infected bacterium to another bacterium. This also refers to the transfer of genes into eukaryotic cells by viruses. This naturally occurring process is routinely employed as a GENE TRANSFER TECHNIQUE.
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.
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
A genus of the family RETROVIRIDAE consisting of non-oncogenic retroviruses that produce multi-organ diseases characterized by long incubation periods and persistent infection. Lentiviruses are unique in that they contain open reading frames (ORFs) between the pol and env genes and in the 3' env region. Five serogroups are recognized, reflecting the mammalian hosts with which they are associated. HIV-1 is the type species.
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 type of FLUORESCENCE SPECTROSCOPY using two FLUORESCENT DYES with overlapping emission and absorption spectra, which is used to indicate proximity of labeled molecules. This technique is useful for studying interactions of molecules and PROTEIN FOLDING.
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.
Agents that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags.
Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
PLANTS, or their progeny, whose GENOME has been altered by GENETIC ENGINEERING.
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.
A plant genus of the family SOLANACEAE. Members contain NICOTINE and other biologically active chemicals; its dried leaves are used for SMOKING.
The in vitro fusion of GENES by RECOMBINANT DNA techniques to analyze protein behavior or GENE EXPRESSION REGULATION, or to merge protein functions for specific medical or industrial uses.
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 class in the phylum CNIDARIA which alternates between polyp and medusa forms during their life cycle. There are over 2700 species in five orders.
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 first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
A tricarbocyanine dye that is used diagnostically in liver function tests and to determine blood volume and cardiac output.
Short, predominantly basic amino acid sequences identified as nuclear import signals for some proteins. These sequences are believed to interact with specific receptors at the NUCLEAR PORE.
Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.
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.
Measurement of the intensity and quality of fluorescence.
A species of CERCOPITHECUS containing three subspecies: C. tantalus, C. pygerythrus, and C. sabeus. They are found in the forests and savannah of Africa. The African green monkey (C. pygerythrus) is the natural host of SIMIAN IMMUNODEFICIENCY VIRUS and is used in AIDS research.
The part of a cell that contains the CYTOSOL and small structures excluding the CELL NUCLEUS; MITOCHONDRIA; and large VACUOLES. (Glick, Glossary of Biochemistry and Molecular Biology, 1990)
Techniques and strategies which include the use of coding sequences and other conventional or radical means to transform or modify cells for the purpose of treating or reversing disease conditions.
A genus of the family PARVOVIRIDAE, subfamily PARVOVIRINAE, which are dependent on a coinfection with helper adenoviruses or herpesviruses for their efficient replication. The type species is Adeno-associated virus 2.
A method used to study the lateral movement of MEMBRANE PROTEINS and LIPIDS. A small area of a cell membrane is bleached by laser light and the amount of time necessary for unbleached fluorescent marker-tagged proteins to diffuse back into the bleached site is a measurement of the cell membrane's fluidity. The diffusion coefficient of a protein or lipid in the membrane can be calculated from the data. (From Segen, Current Med Talk, 1995).
A family of non-enveloped viruses infecting mammals (MASTADENOVIRUS) and birds (AVIADENOVIRUS) or both (ATADENOVIRUS). Infections may be asymptomatic or result in a variety of diseases.
Elements of limited time intervals, contributing to particular results or situations.
Proteins prepared by recombinant DNA technology.
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.
Proteins that originate from plants species belonging to the genus ARABIDOPSIS. The most intensely studied species of Arabidopsis, Arabidopsis thaliana, is commonly used in laboratory experiments.
Proteins found in any species of bacterium.
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 marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts.
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.
Expanded structures, usually green, of vascular plants, characteristically consisting of a bladelike expansion attached to a stem, and functioning as the principal organ of photosynthesis and transpiration. (American Heritage Dictionary, 2d ed)
The basic cellular units of nervous tissue. Each neuron consists of a body, an axon, and dendrites. Their purpose is to receive, conduct, and transmit impulses in the NERVOUS SYSTEM.
The visually perceived property of objects created by absorption or reflection of specific wavelengths of light.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.
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 movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments.
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.
Microscopy in which television cameras are used to brighten magnified images that are otherwise too dark to be seen with the naked eye. It is used frequently in TELEPATHOLOGY.
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.
Components of a cell produced by various separation techniques which, though they disrupt the delicate anatomy of a cell, preserve the structure and physiology of its functioning constituents for biochemical and ultrastructural analysis. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p163)
A technique in which electric pulses of intensity in kilovolts per centimeter and of microsecond-to-millisecond duration cause a temporary loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA.
Any of a variety of procedures which use biomolecular probes to measure the presence or concentration of biological molecules, biological structures, microorganisms, etc., by translating a biochemical interaction at the probe surface into a quantifiable physical signal.
A system of cisternae in the CYTOPLASM of many cells. In places the endoplasmic reticulum is continuous with the plasma membrane (CELL MEMBRANE) or outer membrane of the nuclear envelope. If the outer surfaces of the endoplasmic reticulum membranes are coated with ribosomes, the endoplasmic reticulum is said to be rough-surfaced (ENDOPLASMIC RETICULUM, ROUGH); otherwise it is said to be smooth-surfaced (ENDOPLASMIC RETICULUM, SMOOTH). (King & Stansfield, A Dictionary of Genetics, 4th ed)
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.
Proteins found in plants (flowers, herbs, shrubs, trees, etc.). The concept does not include proteins found in vegetables for which VEGETABLE PROTEINS is available.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in plants.
A phylum of radially symmetrical invertebrates characterized by possession of stinging cells called nematocysts. It includes the classes ANTHOZOA; CUBOZOA; HYDROZOA, and SCYPHOZOA. Members carry CNIDARIAN VENOMS.
A class in the phylum CNIDARIA, comprised mostly of corals and anemones. All members occur only as polyps; the medusa stage is completely absent.
Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs.
Procedures by which protein structure and function are changed or created in vitro by altering existing or synthesizing new structural genes that direct the synthesis of proteins with sought-after properties. Such procedures may include the design of MOLECULAR MODELS of proteins using COMPUTER GRAPHICS or other molecular modeling techniques; site-specific mutagenesis (MUTAGENESIS, SITE-SPECIFIC) of existing genes; and DIRECTED MOLECULAR EVOLUTION techniques to create new genes.
An exotic species of the family CYPRINIDAE, originally from Asia, that has been introduced in North America. They are used in embryological studies and to study the effects of certain chemicals on development.
A stack of flattened vesicles that functions in posttranslational processing and sorting of proteins, receiving them from the rough ENDOPLASMIC RETICULUM and directing them to secretory vesicles, LYSOSOMES, or the CELL MEMBRANE. The movement of proteins takes place by transfer vesicles that bud off from the rough endoplasmic reticulum or Golgi apparatus and fuse with the Golgi, lysosomes or cell membrane. (From Glick, Glossary of Biochemistry and Molecular Biology, 1990)
The infusion of leaves of CAMELLIA SINENSIS (formerly Thea sinensis) as a beverage, the familiar Asian tea, which contains CATECHIN (especially epigallocatechin gallate) and CAFFEINE.
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 protoplasm and plasma membrane of plant, fungal, bacterial or archaeon cells without the CELL WALL.
Chemical reactions effected by light.
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.
Gated transport mechanisms by which proteins or RNA are moved across the NUCLEAR MEMBRANE.
Fluorescence microscopy utilizing multiple low-energy photons to produce the excitation event of the fluorophore. Multiphoton microscopes have a simplified optical path in the emission side due to the lack of an emission pinhole, which is necessary with normal confocal microscopes. Ultimately this allows spatial isolation of the excitation event, enabling deeper imaging into optically thick tissue, while restricting photobleaching and phototoxicity to the area being imaged.
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.
Proteins found in any species of fungus.
Directed modification of the gene complement of a living organism by such techniques as altering the DNA, substituting genetic material by means of a virus, transplanting whole nuclei, transplanting cell hybrids, etc.
That portion of the electromagnetic spectrum in the visible, ultraviolet, and infrared range.
Relatively undifferentiated cells that retain the ability to divide and proliferate throughout postnatal life to provide progenitor cells that can differentiate into specialized cells.
Amino acid sequences found in transported proteins that selectively guide the distribution of the proteins to specific cellular compartments.
A partitioning within cells due to the selectively permeable membranes which enclose each of the separate parts, e.g., mitochondria, lysosomes, etc.
A genetic rearrangement through loss of segments of DNA or RNA, bringing sequences which are normally separated into close proximity. This deletion may be detected using cytogenetic techniques and can also be inferred from the phenotype, indicating a deletion at one specific locus.
Herbaceous biennial plants and their edible bulbs, belonging to the Liliaceae.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
Any spaces or cavities within a cell. They may function in digestion, storage, secretion, or excretion.
Proteins found in any species of virus.
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.
Viral proteins that facilitate the movement of viruses between plant cells by means of PLASMODESMATA, channels that traverse the plant cell walls.
Filamentous proteins that are the main constituent of the thin filaments of muscle fibers. The filaments (known also as filamentous or F-actin) can be dissociated into their globular subunits; each subunit is composed of a single polypeptide 375 amino acids long. This is known as globular or G-actin. In conjunction with MYOSINS, actin is responsible for the contraction and relaxation of muscle.
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.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation.
The usually underground portions of a plant that serve as support, store food, and through which water and mineral nutrients enter the plant. (From American Heritage Dictionary, 1982; Concise Dictionary of Biology, 1990)
Histochemical localization of immunoreactive substances using labeled antibodies as reagents.
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.
Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.
Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.
Family of RNA viruses that infects birds and mammals and encodes the enzyme reverse transcriptase. The family contains seven genera: DELTARETROVIRUS; LENTIVIRUS; RETROVIRUSES TYPE B, MAMMALIAN; ALPHARETROVIRUS; GAMMARETROVIRUS; RETROVIRUSES TYPE D; and SPUMAVIRUS. A key feature of retrovirus biology is the synthesis of a DNA copy of the genome which is integrated into cellular DNA. After integration it is sometimes not expressed but maintained in a latent state (PROVIRUSES).
Techniques where DNA is delivered directly into organelles at high speed using projectiles coated with nucleic acid, shot from a helium-powered gun (gene gun). One of these techniques involves immunization by DNA VACCINES, which delivers DNA-coated gold beads to the epidermis.
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.
The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.
Transport proteins that carry specific substances in the blood or across cell membranes.
Organisms whose GENOME has been changed by a GENETIC ENGINEERING technique.
The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle.
Cellular uptake of extracellular materials within membrane-limited vacuoles or microvesicles. ENDOSOMES play a central role in endocytosis.
A cell line derived from cultured tumor cells.
Slender, cylindrical filaments found in the cytoskeleton of plant and animal cells. They are composed of the protein TUBULIN and are influenced by TUBULIN MODULATORS.
Microscopic threadlike filaments in FUNGI that are filled with a layer of protoplasm. Collectively, the hyphae make up the MYCELIUM.
The rate dynamics in chemical or physical systems.
Pollution prevention through the design of effective chemical products that have low or no toxicity and use of chemical processes that reduce or eliminate the use and generation of hazardous substances.
Semiautonomous, self-reproducing organelles that occur in the cytoplasm of all cells of most, but not all, eukaryotes. Each mitochondrion is surrounded by a double limiting membrane. The inner membrane is highly invaginated, and its projections are called cristae. Mitochondria are the sites of the reactions of oxidative phosphorylation, which result in the formation of ATP. They contain distinctive RIBOSOMES, transfer RNAs (RNA, TRANSFER); AMINO ACYL T RNA SYNTHETASES; and elongation and termination factors. Mitochondria depend upon genes within the nucleus of the cells in which they reside for many essential messenger RNAs (RNA, MESSENGER). Mitochondria are believed to have arisen from aerobic bacteria that established a symbiotic relationship with primitive protoeukaryotes. (King & Stansfield, A Dictionary of Genetics, 4th ed)
Diseases of plants.
Naturally occurring or experimentally induced animal diseases with pathological processes sufficiently similar to those of human diseases. They are used as study models for human diseases.
Proteins found in the nucleus of a cell. Do not confuse with NUCLEOPROTEINS which are proteins conjugated with nucleic acids, that are not necessarily present in the nucleus.
A phylum of photosynthetic EUKARYOTA bearing double membrane-bound plastids containing chlorophyll a and b. They comprise the classical green algae, and represent over 7000 species that live in a variety of primarily aquatic habitats. Only about ten percent are marine species, most live in freshwater.
Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.
Membrane-like channels of cytoplasm connecting adjacent plant cells. Plasmodesmata connect through pores in the CELL WALL and associate with the CYTOSKELETON machinery. They are essential for intercellular transport and communication.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
Microbodies which occur in animal and plant cells and in certain fungi and protozoa. They contain peroxidase, catalase, and allied enzymes. (From Singleton and Sainsbury, Dictionary of Microbiology and Molecular Biology, 2nd ed)
The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability.
Theoretical representations that simulate the behavior or activity of biological processes or diseases. For disease models in living animals, DISEASE MODELS, ANIMAL is available. Biological models include the use of mathematical equations, computers, and other electronic equipment.
Proteins obtained from the species SACCHAROMYCES CEREVISIAE. The function of specific proteins from this organism are the subject of intense scientific interest and have been used to derive basic understanding of the functioning similar proteins in higher eukaryotes.
Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy.
Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.
Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.
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.
Mutagenesis where the mutation is caused by the introduction of foreign DNA sequences into a gene or extragenic sequence. This may occur spontaneously in vivo or be experimentally induced in vivo or in vitro. Proviral DNA insertions into or adjacent to a cellular proto-oncogene can interrupt GENETIC TRANSLATION of the coding sequences or interfere with recognition of regulatory elements and cause unregulated expression of the proto-oncogene resulting in tumor formation.
A 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.
The developmental history of specific differentiated cell types as traced back to the original STEM CELLS in the embryo.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.
The entity of a developing mammal (MAMMALS), generally from the cleavage of a ZYGOTE to the end of embryonic differentiation of basic structures. For the human embryo, this represents the first two months of intrauterine development preceding the stages of the FETUS.
Discrete concentrations of energy, apparently massless elementary particles, that move at the speed of light. They are the unit or quantum of electromagnetic radiation. Photons are emitted when electrons move from one energy state to another. (From Hawley's Condensed Chemical Dictionary, 11th ed)
A gene silencing phenomenon whereby specific dsRNAs (RNA, DOUBLE-STRANDED) trigger the degradation of homologous mRNA (RNA, MESSENGER). The specific dsRNAs are processed into SMALL INTERFERING RNA (siRNA) which serves as a guide for cleavage of the homologous mRNA in the RNA-INDUCED SILENCING COMPLEX. DNA METHYLATION may also be triggered during this process.
A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5.
Refers to animals in the period of time just after birth.
The tendency of a gas or solute to pass from a point of higher pressure or concentration to a point of lower pressure or concentration and to distribute itself throughout the available space. Diffusion, especially FACILITATED DIFFUSION, is a major mechanism of BIOLOGICAL TRANSPORT.
Commonly observed structural components of proteins formed by simple combinations of adjacent secondary structures. A commonly observed structure may be composed of a CONSERVED SEQUENCE which can be represented by a CONSENSUS SEQUENCE.
Cells contained in the bone marrow including fat cells (see ADIPOCYTES); STROMAL CELLS; MEGAKARYOCYTES; and the immediate precursors of most blood cells.
Recombinases that insert exogenous DNA into the host genome. Examples include proteins encoded by the POL GENE of RETROVIRIDAE and also by temperate BACTERIOPHAGES, the best known being BACTERIOPHAGE LAMBDA.
A species of nematode that is widely used in biological, biochemical, and genetic studies.
A thin layer of cells forming the outer integument of seed plants and ferns. (Random House Unabridged Dictionary, 2d ed)
Any of the processes by which cytoplasmic or intercellular factors influence the differential control of gene action in bacteria.
The integration of exogenous DNA into the genome of an organism at sites where its expression can be suitably controlled. This integration occurs as a result of homologous recombination.
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.
Enzymes that oxidize certain LUMINESCENT AGENTS to emit light (PHYSICAL LUMINESCENCE). The luciferases from different organisms have evolved differently so have different structures and substrates.
An antioxidant flavonoid, occurring especially in woody plants as both (+)-catechin and (-)-epicatechin (cis) forms.
Deletion of sequences of nucleic acids from the genetic material of an individual.
A genus of plant viruses in the family FLEXIVIRIDAE, that cause mosaic and ringspot symptoms. Transmission occurs mechanically. Potato virus X is the type species.
The movement of cells from one location to another. Distinguish from CYTOKINESIS which is the process of dividing the CYTOPLASM of a cell.
Self-replicating cytoplasmic organelles of plant and algal cells that contain pigments and may synthesize and accumulate various substances. PLASTID GENOMES are used in phylogenetic studies.
Cytoplasmic vesicles formed when COATED VESICLES shed their CLATHRIN coat. Endosomes internalize macromolecules bound by receptors on the cell surface.
Cell lines whose original growing procedure consisted being transferred (T) every 3 days and plated at 300,000 cells per plate (J Cell Biol 17:299-313, 1963). Lines have been developed using several different strains of mice. Tissues are usually fibroblasts derived from mouse embryos but other types and sources have been developed as well. The 3T3 lines are valuable in vitro host systems for oncogenic virus transformation studies, since 3T3 cells possess a high sensitivity to CONTACT INHIBITION.
The relationships of groups of organisms as reflected by their genetic makeup.
A CELL LINE derived from the kidney of the African green (vervet) monkey, (CERCOPITHECUS AETHIOPS) used primarily in virus replication studies and plaque assays.
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.
Biologically functional sequences of DNA chemically synthesized in vitro.
Cells derived from the BLASTOCYST INNER CELL MASS which forms before implantation in the uterine wall. They retain the ability to divide, proliferate and provide progenitor cells that can differentiate into specialized cells.
The injection of very small amounts of fluid, often with the aid of a microscope and microsyringes.
The developmental entity of a fertilized egg (ZYGOTE) in animal species other than MAMMALS. For chickens, use CHICK EMBRYO.
A pyridoxal-phosphate protein that catalyzes the alpha-decarboxylation of L-glutamic acid to form gamma-aminobutyric acid and carbon dioxide. The enzyme is found in bacteria and in invertebrate and vertebrate nervous systems. It is the rate-limiting enzyme in determining GAMMA-AMINOBUTYRIC ACID levels in normal nervous tissues. The brain enzyme also acts on L-cysteate, L-cysteine sulfinate, and L-aspartate. EC
Interruption or suppression of the expression of a gene at transcriptional or translational levels.
The membrane system of the CELL NUCLEUS that surrounds the nucleoplasm. It consists of two concentric membranes separated by the perinuclear space. The structures of the envelope where it opens to the cytoplasm are called the nuclear pores (NUCLEAR PORE).
Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.
Plants or plant parts which are harmful to man or other animals.
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.
Specific particles of membrane-bound organized living substances present in eukaryotic cells, such as the MITOCHONDRIA; the GOLGI APPARATUS; ENDOPLASMIC RETICULUM; LYSOSOMES; PLASTIDS; and VACUOLES.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Mutant mice homozygous for the recessive gene "nude" which fail to develop a thymus. They are useful in tumor studies and studies on immune responses.
A strain of albino rat used widely for experimental purposes because of its calmness and ease of handling. It was developed by the Sprague-Dawley Animal Company.
Membrane proteins whose primary function is to facilitate the transport of molecules across a biological membrane. Included in this broad category are proteins involved in active transport (BIOLOGICAL TRANSPORT, ACTIVE), facilitated transport and ION CHANNELS.
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)
Camellia sinensis L. (formerly Thea sinensis) is an evergreen Asiatic shrub of the THEACEAE family. The infusion of leaves of this plant is used as Oriental TEA which contains CAFFEINE; THEOPHYLLINE; and epigallocatechin gallate.
Luciferases from FIREFLIES, usually Photinus, that oxidizes FIREFLY LUCIFERIN to cause emission of PHOTONS.
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.
Different forms of a protein that may be produced from different GENES, or from the same gene by ALTERNATIVE SPLICING.
Thin structures that encapsulate subcellular structures or ORGANELLES in EUKARYOTIC CELLS. They include a variety of membranes associated with the CELL NUCLEUS; the MITOCHONDRIA; the GOLGI APPARATUS; the ENDOPLASMIC RETICULUM; LYSOSOMES; PLASTIDS; and VACUOLES.
Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies.
Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.
An electrophysiologic technique for studying cells, cell membranes, and occasionally isolated organelles. All patch-clamp methods rely on a very high-resistance seal between a micropipette and a membrane; the seal is usually attained by gentle suction. The four most common variants include on-cell patch, inside-out patch, outside-out patch, and whole-cell clamp. Patch-clamp methods are commonly used to voltage clamp, that is control the voltage across the membrane and measure current flow, but current-clamp methods, in which the current is controlled and the voltage is measured, are also used.
A basic element found in nearly all organized tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes.
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.
A type VI intermediate filament protein expressed mostly in nerve cells where it is associated with the survival, renewal and mitogen-stimulated proliferation of neural progenitor cells.
Characteristic restricted to a particular organ of the body, such as a cell type, metabolic response or expression of a particular protein or antigen.
Non-invasive imaging of cells that have been labeled non-destructively, such as with nanoemulsions or reporter genes that can be detected by molecular imaging, to monitor their location, viability, cell lineage expansion, response to drugs, movement, or other behaviors in vivo.

Mitochondrial depolarization accompanies cytochrome c release during apoptosis in PC6 cells. (1/19912)

Cytochrome c is released from mitochondria into the cytosol in cells undergoing apoptosis. The temporal relationship between cytochrome c release and loss of mitochondrial membrane potential was monitored by laser-scanning confocal microscopy in single living pheochromocytoma-6 cells undergoing apoptosis induced by staurosporine. Mitochondrial membrane potential monitored by tetramethylrhodamine methyl ester decreased abruptly in individual cells from 2 to 7 h after treatment with staurosporine. Depolarization was accompanied by cytochrome c release documented by release of transfected green fluorescent protein-tagged cytochrome c in these cells. The results show that mitochondrial depolarization accompanies cytochrome c release in pheochromocytoma-6 cells undergoing apoptosis.  (+info)

The dually acylated NH2-terminal domain of gi1alpha is sufficient to target a green fluorescent protein reporter to caveolin-enriched plasma membrane domains. Palmitoylation of caveolin-1 is required for the recognition of dually acylated g-protein alpha subunits in vivo. (2/19912)

Here we investigate the molecular mechanisms that govern the targeting of G-protein alpha subunits to the plasma membrane. For this purpose, we used Gi1alpha as a model dually acylated G-protein. We fused full-length Gi1alpha or its extreme NH2-terminal domain (residues 1-32 or 1-122) to green fluorescent protein (GFP) and analyzed the subcellular localization of these fusion proteins. We show that the first 32 amino acids of Gi1alpha are sufficient to target GFP to caveolin-enriched domains of the plasma membrane in vivo, as demonstrated by co-fractionation and co-immunoprecipitation with caveolin-1. Interestingly, when dual acylation of this 32-amino acid domain was blocked by specific point mutations (G2A or C3S), the resulting GFP fusion proteins were localized to the cytoplasm and excluded from caveolin-rich regions. The myristoylated but nonpalmitoylated (C3S) chimera only partially partitioned into caveolin-containing fractions. However, both nonacylated GFP fusions (G2A and C3S) no longer co-immunoprecipitated with caveolin-1. Taken together, these results indicate that lipid modification of the NH2-terminal of Gi1alpha is essential for targeting to its correct destination and interaction with caveolin-1. Also, a caveolin-1 mutant lacking all three palmitoylation sites (C133S, C143S, and C156S) was unable to co-immunoprecipitate these dually acylated GFP-G-protein fusions. Thus, dual acylation of the NH2-terminal domain of Gi1alpha and palmitoylation of caveolin-1 are both required to stabilize and perhaps regulate this reciprocal interaction at the plasma membrane in vivo. Our results provide the first demonstration of a functional role for caveolin-1 palmitoylation in its interaction with signaling molecules.  (+info)

Properties of filament-bound myosin light chain kinase. (3/19912)

Myosin light chain kinase binds to actin-containing filaments from cells with a greater affinity than to F-actin. However, it is not known if this binding in cells is regulated by Ca2+/calmodulin as it is with F-actin. Therefore, the binding properties of the kinase to stress fibers were examined in smooth muscle-derived A7r5 cells. Full-length myosin light chain kinase or a truncation mutant lacking residues 2-142 was expressed as chimeras containing green fluorescent protein at the C terminus. In intact cells, the full-length kinase bound to stress fibers, whereas the truncated kinase showed diffuse fluorescence in the cytoplasm. After permeabilization with saponin, the fluorescence from the truncated kinase disappeared, whereas the fluorescence of the full-length kinase was retained on stress fibers. Measurements of fluorescence intensities and fluorescence recovery after photobleaching of the full-length myosin light chain kinase in saponin-permeable cells showed that Ca2+/calmodulin did not dissociate the kinase from these filaments. However, the filament-bound kinase was sufficient for Ca2+-dependent phosphorylation of myosin regulatory light chain and contraction of stress fibers. Thus, dissociation of myosin light chain kinase from actin-containing thin filaments is not necessary for phosphorylation of myosin light chain in thick filaments. We note that the distance between the N terminus and the catalytic core of the kinase is sufficient to span the distance between thin and thick filaments.  (+info)

A fluorescent orthotopic bone metastasis model of human prostate cancer. (4/19912)

Here, we report a fluorescent spontaneous bone metastatic model of human prostate cancer developed by surgical orthotopic implantation of green fluorescent protein (GFP)-expressing prostate cancer tissue. Human prostate cancer PC-3 cells were transduced with the pLEIN expression retroviral vector containing the enhanced GFP and neomycin resistance genes. Stable GFP high-expression PC-3 clones were selected in vitro with G418, which were then combined and injected s.c. in nude mice. For metastasis studies, fragments of a single highly fluorescent s.c. growing tumor were implanted by surgical orthotopic implantation in the prostate of a series of nude mice. Subsequent micrometastases and metastases were visualized by GFP fluorescence throughout the skeleton, including the skull, rib, pelvis, femur, and tibia The central nervous system, including the brain and spinal cord, was also involved with tumor, as visualized by GFP fluorescence. Systemic organs, including the lung, plural membrane, liver, kidney, and adrenal gland, also had fluorescent metastases. The metastasis pattern in this model reflects the bone and other metastatic sites of human prostate cancer. Thus, this model should be very useful for the study and development of treatment for metastatic androgen-independent prostate cancer.  (+info)

Role of cytokine signaling molecules in erythroid differentiation of mouse fetal liver hematopoietic cells: functional analysis of signaling molecules by retrovirus-mediated expression. (5/19912)

Erythropoietin (EPO) and its cell surface receptor (EPOR) play a central role in proliferation, differentiation, and survival of erythroid progenitors. Signals induced by EPO have been studied extensively by using erythroid as well as nonerythroid cell lines, and various controversial results have been reported as to the role of signaling molecules in erythroid differentiation. Here we describe a novel approach to analyze the EPO signaling by using primary mouse fetal liver hematopoietic cells to avoid possible artifacts due to established cell lines. Our strategy is based on high-titer retrovirus vectors with a bicistronic expression system consisting of an internal ribosome entry site (IRES) and green fluorescent protein (GFP). By placing the cDNA for a signaling molecule in front of IRES-GFP, virus-infected cells can be viably sorted by fluorescence-activated cell sorter, and the effect of expression of the signaling molecule can be assessed. By using this system, expression of cell-survival genes such as Bcl-2 and Bcl-XL was found to enhance erythroid colony formation from colony-forming unit-erythroid (CFU-E) in response to EPO. However, their expression was not sufficient for erythroid colony formation from CFU-E alone, indicating that EPO induces signals for erythroid differentiation. To examine the role of EPOR tyrosine residues in erythroid differentiation, we introduced a chimeric EGFR-EPOR receptor, which has the extracellular domain of the EGF receptor and the intracellular domain of the EPOR, as well as a mutant EGFR-EPOR in which all the cytoplasmic tyrosine residues are replaced with phenylalanine, and found that tyrosine residues of EPOR are essential for erythroid colony formation from CFU-E. We further analyzed the function of the downstream signaling molecules by expressing modified signaling molecules and found that both JAK2/STAT5 and Ras, two major signaling pathways activated by EPOR, are involved in full erythroid differentiation.  (+info)

Nuclear translocation of green fluorescent protein-nuclear factor kappaB with a distinct lag time in living cells. (6/19912)

A highly fluorescent mutant form of the green fluorescent protein (GFP) has been fused to the human nuclear factor kappaB (NF-kappaB) p50 and p105 (p50/IkappaB gamma), a precursor protein of NF-kappaB p50. GFP-p50 and GFP-p105 were expressed in monkey COS-7 cells and human HeLa cells. Translocation of these chimeric proteins was observed by confocal laser scanning microscopy. GFP-p50 (without IkappaB gamma) in the transfected cells resided in the nucleus. On the other hand, GFP-p105 (GFP-p50 with IkappaB gamma) localized only in the cytoplasm before stimulation and translocated to the nucleus with stimulant specificity similar to that of native NF-kappaB/IkappaB. In addition, the translocation of NF-kappaB to the nucleus had a distinct lag time (a quiescent time) in the target cells. The lag time lasted 10-20 min after stimulation with hydrogen peroxide or tumor necrosis factor alpha. It was suggested that this might be due to the existence of a limiting step where NF-kappaB is released from NF-kappaB/IkappaB by the proteasome.  (+info)

Microtubule-dependent plus- and minus end-directed motilities are competing processes for nuclear targeting of adenovirus. (7/19912)

Adenovirus (Ad) enters target cells by receptor-mediated endocytosis, escapes to the cytosol, and then delivers its DNA genome into the nucleus. Here we analyzed the trafficking of fluorophore-tagged viruses in HeLa and TC7 cells by time-lapse microscopy. Our results show that native or taxol-stabilized microtubules (MTs) support alternating minus- and plus end-directed movements of cytosolic virus with elementary speeds up to 2.6 micrometer/s. No directed movement was observed in nocodazole-treated cells. Switching between plus- and minus end-directed elementary speeds at frequencies up to 1 Hz was observed in the periphery and near the MT organizing center (MTOC) after recovery from nocodazole treatment. MT-dependent motilities allowed virus accumulation near the MTOC at population speeds of 1-10 micrometer/min, depending on the cell type. Overexpression of p50/dynamitin, which is known to affect dynein-dependent minus end-directed vesicular transport, significantly reduced the extent and the frequency of minus end-directed migration of cytosolic virus, and increased the frequency, but not the extent of plus end-directed motility. The data imply that a single cytosolic Ad particle engages with two types of MT-dependent motor activities, the minus end- directed cytoplasmic dynein and an unknown plus end- directed activity.  (+info)

Estimation of the number of alpha-helical and beta-strand segments in proteins using circular dichroism spectroscopy. (8/19912)

A simple approach to estimate the number of alpha-helical and beta-strand segments from protein circular dichroism spectra is described. The alpha-helix and beta-sheet conformations in globular protein structures, assigned by DSSP and STRIDE algorithms, were divided into regular and distorted fractions by considering a certain number of terminal residues in a given alpha-helix or beta-strand segment to be distorted. The resulting secondary structure fractions for 29 reference proteins were used in the analyses of circular dichroism spectra by the SELCON method. From the performance indices of the analyses, we determined that, on an average, four residues per alpha-helix and two residues per beta-strand may be considered distorted in proteins. The number of alpha-helical and beta-strand segments and their average length in a given protein were estimated from the fraction of distorted alpha-helix and beta-strand conformations determined from the analysis of circular dichroism spectra. The statistical test for the reference protein set shows the high reliability of such a classification of protein secondary structure. The method was used to analyze the circular dichroism spectra of four additional proteins and the predicted structural characteristics agree with the crystal structure data.  (+info)

Structural investigations on green fluorescent protein variants and the adenylyl cyclase associated protein [Elektronische Ressource] / Dorota Ksiazek : Technische Universität München Institut für Organische Chemie und Biochemie Max-Planck-Institut für Biochemie Abteilung Strukturforschung Biologische NMR-Arbeitsgruppe Structural Investigations on Green Fluorescent Protein Variants and the Adenylyl Cyclase-Associated Protein Dorota Ksiazek Vollständiger Abdruck der von der Fakultät für Chemie der Technischen Universität München zur
Aaron Kolb // Brandt Lab // Publications // Dec 06 2003. PubMed ID: 14656463. Author(s): Kolb AW, Brandt CR. Enhanced isolation of low frequency herpes simplex virus recombinants using green-fluorescent protein and FACS. J Virol Methods. 2004 Jan;115(1):73-81. Journal: Journal Of Virological Methods, Volume 115, Issue 1, Jan 2004. The generation of recombinant herpes simplex virus to study the effect of engineered mutations on viral biology relies on the isolation of recombinants from a mixed population of viruses following a marker transfer procedure. Currently, the E. coli lacZ or green-fluorescent protein (GFP) genes are most frequently used as markers for isolation and isolation of recombinants relies on visual screening of plaques. Alternatively, novel restriction site changes can be inserted into a gene followed by screening of individual plaques for the novel change. These methods are inefficient when the frequency of recombinants in the pool of viruses is low. Using GFP as a selection ...
TY - JOUR. T1 - Green fluorescent protein mutant as label in homogeneous assays for biomolecules. AU - Deo, Sapna K.. AU - Daunert, Sylvia. PY - 2001/2/1. Y1 - 2001/2/1. N2 - The green fluorescent protein (GFP) and its mutants have been extensively used to study various cellular processes and, more recently, as labels in binding assays. We have employed a mutant of GFP, an enhanced GFP (EGFP), in the development of homogeneous assays for biotin and cortisol. To demonstrate the feasibility of using EGFP as a label with different kinds of binders in the development of homogeneous assays, we employed the biotin-avidin and an antigen-antibody as the binding pairs. Biotin and cortisol were chemically conjugated to EGFP. A quenching of fluorescence intensity of EGFP was observed upon binding of avidin to the EGFP-biotin conjugate. The percentage fluorescence quenching observed decreased as the concentration of free biotin in the sample increased due to the fewer binding sites on avidin available for ...
Molecular Plant-Microbe Interactions 10:394-400...Continual Green-Fluorescent Protein Monitoring of Cauliflower Mosaic Virus 35S Promoter Activity in Nematode-Induced Feeding Cells in Arabidopsis thaliana...
Read this Science Presentation or Speech and over 30,000 other research documents. Introduction to Green Fluorescent Protein (gfp).. Introduction to Green Fluorescent Protein (GFP) The green fluorescent protein (GFP) obtained from the jellyfish Aequorea victoria has become one of the most widely researched proteins in biochemistry and cell biology, In early 1960s researcher named Osamu Shimomura set out to find the reason for this luminescence , after harvesting...
OV-IA82Δ113 and OV-IA82Δ116 viruses were constructed by infecting OFTu cells (in T25 flasks) at a multiplicity of infection (MOI) of 1.0 with wild type OV-IA 82 for 3 hours and subsequently transfecting the cells with 10 μg of pSVP-113LF-EGFP-113RF and pSVP-116LF-EGFP-116RF transfer vectors by standard in vivo recombination protocols [28, 29]. Transfections were carried out using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturers instructions.. Viruses were harvested 48 h pi by scraping infected/transfected OFTu cells into sterile 15 ml conical tubes. The cell suspensions were vortexed, frozen/thawed 3 times, and then centrifuged at 1000 rpm, for 10 min at 4°C (Eppendorf Centrifuge 5810R, 15 amp version, Hamburg, Germany). The supernatant (viruses) were transferred into 2 ml cryogenic vials (Corning, NY, USA) and stored at -80°C for future use.. In order to select and purify recombinant viruses, limited dilution and plaque assays were performed. 3 ×104 of ...
TY - JOUR. T1 - FMDV replicons encoding green fluorescent protein are replication competent. AU - Tulloch, Fiona. AU - Pathania, Uday. AU - Luke, Garry A.. AU - Nicholson, John. AU - Stonehouse, Nicola J.. AU - Rowlands, David J.. AU - Jackson, Terry. AU - Tuthill, Toby. AU - Haas, Juergen. AU - Lamond, Angus I.. AU - Ryan, Martin D.. PY - 2014/12/1. Y1 - 2014/12/1. N2 - The study of replication of viruses that require high bio-secure facilities can be accomplished with less stringent containment using non-infectious replicon systems. The FMDV replicon system (pT7rep) reported by Mclnerney et al. (2000) was modified by the replacement of sequences encoding chloramphenicol acetyl-transferase (CAT) with those encoding a functional (L pro) linked to a bi-functional fluorescent/antibiotic resistance fusion protein (green fluorescent protein/puromycin resistance, [GFP-PAC]). Cells were transfected with replicon-derived transcript RNA and GFP fluorescence quantified. Replication of transcript RNAs ...
The monoclonal Anti-GFP-HRP antibody facilitates the fast and convenient detection of GFP and GFP derivatives or GFP-tagged fusion proteins in Western blot or ELISA applications. Conjugation to horseradish peroxidase (HRP) simplifies Western blot or ELISA analysis as incubation with secondary antibodies becomes dispensable. After incubation the Anti-GFP-HRP antibody can be directly detected using commercially available chemiluminescent reagents. The monoclonal Anti-GFP antibody specifically reconizes wild-type GFP from Aequorea victoria and derivatives, e.g., EGFP, CFP, YFP, and BFP. - 中国
Expression and Purification of recombinant Green Fluorescent Protein ABSTRACT: The purpose of this experiment was to determine if a His-6 tagged recombinant form of Green Fluorescent Protein could be expressed in a pRSETA vector of E. Coli. This was determined through multiple procedures beginning with purifying the sample with Ni +2 agarose chromatography which showcased the relative fluorescent activity of the samples, which elution sample two (E2) had approximately 100,592.2 RFU/mg . The yield of total protein was found by use of a Bradford Assay and a standard curve. The purity of the GFP was determined by comparing the intensity of bands that appeared at around 31.4 kDa (the molecular weight of rGFP) to a molecular weight ladder on an SDS-PAGE gel. The Western Blot test, utilizing a nitrocellulous membrane, confirmed the expression of rGFP. The Western Blot confirmed that the correct bands were analyzed in the SDS-PAGE gel which E3 had an estimated purity of 0.4, indicating a yield of ...
Goat Polyclonal GFP antibody for FLISA, ICC, FACS, IHC (fro), IF, PLA, WB. Published in 7 Pubmed References. Order this anti-GFP antibody. | Product number ABIN151299
TY - JOUR. T1 - A transgenic rat expressing green fluorescent protein (GFP) in peripheral nerves provides a new hindlimb model for the study of nerve injury and regeneration. AU - Moore, Amy M.. AU - Borschel, Gregory H.. AU - Santosa, Katherine B.. AU - Flagg, Eric R.. AU - Tong, Alice Y.. AU - Kasukurthi, Rahul. AU - Newton, Piyaraj. AU - Yan, Ying. AU - Hunter, Daniel A.. AU - Johnson, Philip J.. AU - Mackinnon, Susan E.. PY - 2012/2/1. Y1 - 2012/2/1. N2 - Background: In order to evaluate nerve regeneration in clinically relevant hindlimb surgical paradigms not feasible in fluorescent mice models, we developed a rat that expresses green fluorescent protein (GFP) in neural tissue. Methods: Transgenic Sprague-Dawley rat lines were created using pronuclear injection of a transgene expressing GFP under the control of the thy1 gene. Nerves were imaged under fluorescence microscopy and muscles were imaged with confocal microscopy to determine GFP expression following sciatic nerve crush, ...
Anti-GFP antibody conjugated to Biotin [LGB-1] validated for WB. Referenced in 1 publication. Immunogen corresponding to recombinant full length protein
As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards. The RCSB PDB also provides a variety of tools and resources. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.
Lack of expression of dystrophin leads to degeneration of muscle fibers and infiltration of connective and adipose tissue. Cell transplantation therapy has been proposed as a treatment for intractable muscle degenerative disorders. Several reports have demonstrated the ability of bone-marrow derived cells (BMDC) to contribute to non-haematopoietic tissues including epithelium, heart, liver, skeletal muscle and brain following transplantation by means of fusion and reprogramming. A key issue is the extent to which fusion and reprogramming can occur in vivo, particularly under conditions of myogenic deterioration.To investigate the therapeutic potential of bone marrow transplantation in monogenetic myopathy, green fluorescent protein-positive (GFP+) bone marrow cells were transplanted into non-irradiated c-kit receptor-deficient (W⁴¹) mdx mice. This model allows BMDC reconstitution in the absence of irradiation induced myeloablation. We provide the first report of BMDC fusion in a ...
Aims Cells derived from the stroma vascular fraction (SVF) of mouse adipose tissue can spontaneously give rise to rare, functional, cardiac-like cells in vitro. This study aimed to improve the production of adipose-derived cardiomyogenic cells (AD-CMG), to characterize them and to assess their cardiac fate and functional outcomes after their administration in a mouse model of acute myocardial infarction. Methods and results The culture process optimized to improve in vitro cardiac specification consisted of a primary culture of murine SVF cells in semi-solid methylcellulose medium, a selection of AD-CMG cell clusters, and a secondary culture and expansion in BHK21 medium. AD-CMG cells were CD29+, CD31−, CD34−, CD44+, CD45−, CD81+, CD90−, CD117−, and Flk-1− and expressed several cardiac contractile proteins. After 1, 2, and 4 weeks of their injection in mice having acute myocardial infarction, a strong presence of green fluorescent protein-positive cells was identified by ...
The recruitment of monocytes and their differentiation into macrophages at sites of inflammation are key events in determining the outcome of the inflammatory response and initiating the return to tissue homeostasis. To study monocyte trafficking and macrophage differentiation in vivo, we have generated a novel transgenic reporter mouse expressing a green fluorescent protein (GFP) under the control of the human CD68 promoter. CD68-GFP mice express high levels of GFP in both monocyte and embryo-derived tissue resident macrophages in adult animals. The human CD68 promoter drives GFP expression in all CD115(+) monocytes of adult blood, spleen, and bone marrow; we took advantage of this to directly compare the trafficking of bone marrow-derived CD68-GFP monocytes to that of CX3CR1(GFP) monocytes in vivo using a sterile zymosan peritonitis model. Unlike CX3CR1(GFP) monocytes, which downregulate GFP expression on differentiation into macrophages in this model, CD68-GFP monocytes retain high-level GFP
Novel fluorescent tools such as green fluorescent protein analogs and Fluorogen Activating Proteins (FAPs) are useful in biological imaging to track protein dynamics in real-time with low fluorescence background. evolution experiments of both VH-VL M8 and M8VL, led us to rationally design tandem, covalent homodimers of M8VL domains joined by a flexible linker that have a higher affinity for DIR and great quantum yield. The introduction of fluorescent systems has revolutionized mobile imaging and molecular biology, as well as the electricity of encoded fluorescent proteins, such as for example green fluorescent proteins (GFP), for the recognition of particular proteins appealing can be well recorded (1). Theres a dependence on extra still, well-characterized tools offering real-time, high signal-to-noise fluorescence and demonstrate high fluorescence quantum produce (?f), photo-stability, and a wide spectral range. Fluorogen Activating Protein (FAPs) are section of book, immunoglobulin-based, ...
Aequoria victoria green fluorescent protein (GFP) is a revolutionary molecular biology tool because of its spontaneous peptide backbone cyclization and chromophore formation from residues Ser65, Tyr66, and Gly67. Here we use structure-based design, comprehensive targeted mutagenesis, and high-resolution crystallography to probe the significant functional role of conserved Arg96 (R96) in chromophore maturation. The R96M GFP variant, in which the R96M side chain is similar in volume but lacks the R96 positive charge, exhibits dramatically slower chromophore maturation kinetics (from hours to months). Comparison of the precyclized conformation of the chromophore-forming residues with the mature R96M chromophore reveals a similar Y66 conformer, contrary to the large Y66 conformational change previously defined in the slowly maturing R96A variant [Barondeau, D. P., Putnam, C. D., Kassmann, C. J., Tainer, J. A., and Getzoff, E. D. (2003) Proc. Natl. Acad. Sci. U.S.A. 100, 12111-12116]. Comprehensive ...
TY - JOUR. T1 - Studying cytoskeletal dynamics in living cells using green fluorescent protein. AU - Yoon, Yisang. AU - Pitts, Kelly. AU - McNiven, Mark. PY - 2002/7/4. Y1 - 2002/7/4. N2 - Microfilaments, intermediate filaments, and microtubules are three major cytoskeletal systems providing cells with stability to maintain proper shape. Although the word cytoskeleton implicates rigidity, it is quite dynamic exhibiting constant changes within cells. In addition to providing cell stability, it participates in a variety of essential and dynamic cellular processes including cell migration, cell division, intracellular transport, vesicular trafficking, and organelle morphogenesis. During the past eight years since the green fluorescent protein (GFP) was first used as a marker for the exogenous gene expression, it has been an especially booming era for live cell observations of intracellular movement of many proteins. Because of the dynamic behavior of the cytoskeleton in the cell, GFP has ...
References for The Development of New FPs 15. Shaner , N C, Patterson, G H, & Davidson, M W. (2007). Advances in fluorescent protein technology . Journal of Cell Science, 120. PMID: 18057027 16. Shaner, N C, Steinbach, P A & Tsien, R Y. (2005). A guide to choosing fluorescent proteins. Nature Methods, 2, PMID: 16299475 17. Ai, H, Shaner, N C, Cheng, Z, Tsien, R Y & Campbell, R E. (2007). Exploration of new chromophore structures leads to the identification of improved blue fluorescent proteins. Biochemistry, 46, PMID: 17444659 18. Kremers, G J, Goedhart, J, van den Heuvel, D J, Gerritsen, H C. & Gadella, T W J. (2007). Improved green and blue fluorescent proteins for expression in bacteria and mammalian cells. Biochemistry, 46, PMID: 17323929 19. Cubitt, A B., Heim, R, Adams, S R, Boyd, A E, Gross, L A & Tsien, R Y. (1995). Understanding, improving and using green fluorescent proteins. Trends Biochemical Science, 20, PMID: 8578587 20. Rizzo, M A, Springer, G H, Granada, B & Piston, D W. (2004). ...
References for The Development of New FPs 15. Shaner , N C, Patterson, G H, & Davidson, M W. (2007). Advances in fluorescent protein technology . Journal of Cell Science, 120. PMID: 18057027 16. Shaner, N C, Steinbach, P A & Tsien, R Y. (2005). A guide to choosing fluorescent proteins. Nature Methods, 2, PMID: 16299475 17. Ai, H, Shaner, N C, Cheng, Z, Tsien, R Y & Campbell, R E. (2007). Exploration of new chromophore structures leads to the identification of improved blue fluorescent proteins. Biochemistry, 46, PMID: 17444659 18. Kremers, G J, Goedhart, J, van den Heuvel, D J, Gerritsen, H C. & Gadella, T W J. (2007). Improved green and blue fluorescent proteins for expression in bacteria and mammalian cells. Biochemistry, 46, PMID: 17323929 19. Cubitt, A B., Heim, R, Adams, S R, Boyd, A E, Gross, L A & Tsien, R Y. (1995). Understanding, improving and using green fluorescent proteins. Trends Biochemical Science, 20, PMID: 8578587 20. Rizzo, M A, Springer, G H, Granada, B & Piston, D W. (2004). ...
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
The assessment of how many cells have been successfully transfected or transduced in a cell population is a basic and critical evaluation parameter in many cell and molecular biology labs. Commonly, the cells of interest are transfected or transduced with a construct that results in the expression of a fluorescent protein (FP) reporter, such as GFP.
TY - JOUR. T1 - Chapter 1. T2 - Biophysics of the Green Fluorescent Protein. AU - Prendergast, F. G.. PY - 1998/1/1. Y1 - 1998/1/1. N2 - It is almost certainly a truism that the interpretation of the fluorescence of a protein matrix-embedded chromophore in terms of the physicochemical character of its environment requires that the tertiary structure of the protein be known to high resolution. This reality derives from the complexity of the photophysics of most fluorescent molecules-complexity that reveals the imperfections of available theory. The accuracy of these dicta is highlighted by the biophysical properties of the green fluorescent protein (GFP) now being so elegantly elucidated from the application of X-ray crystallography, ultrafast optical spectroscopy, and site-specific mutagenesis. Given the apparent malleability of the GFP sequence and the sensitivity of the chromophores photophysics to a broad spectrum of physicochemical factors, it is inevitable that additional useful and ...
The green fluorescent protein (GFP) is the most commonly used reporter protein for monitoring gene expression and protein localization in a variety of living and fixed cells, including not only prokaryotes, but eukaryotes also, e. EGFP inhibited both cell nest and growth development, and activated cell loss of life in Ku80-lacking hamster cells, i.y., xrs-6 cells. In addition, Ku80 attenuated EGFP-induced cytotoxicity in the xrs-6 cells. No EGFP-induced cytotoxicity was noticed in the NHEJ primary proteins XRCC4-lacking hamster cells, i.y., XR-1 cells. Furthermore, Substantially enhanced X-ray-induced cytotoxicity in the xrs-6 cells EGFP. These outcomes recommend that Ku80 has a essential function in the story NHEJ-independent protection system against EGFP-induced cytotoxicity. Extreme care should end up being used in taking into consideration of the potential impact by the tension response system, specifically, the Ku80-reliant reduction system of EGFP-induced cytotoxicity, getting turned on, ...
TY - JOUR. T1 - DNA sequence-enabled reassembly of the green fluorescent protein. AU - Stains, Cliff I.. AU - Porter, Jason R.. AU - Ooi, Aik T.. AU - Segal, David J.. AU - Ghosh, Indraneel. PY - 2005/8/10. Y1 - 2005/8/10. N2 - We describe a general methodology for the direct detection of DNA by the design of a split-protein system that reassembles to form an active complex only in the presence of a targeted DNA sequence. This approach, called SEquence Enabled Reassembly (SEER) of proteins, combines the ability to rationally dissect proteins to construct oligomerization-dependent protein reassembly systems and the availability of DNA binding Cys2-His2 zinc-finger motifs for the recognition of specific DNA sequences. We demonstrate the feasibility of the SEER approach utilizing the split green fluorescent protein appended to appropriate zinc fingers, such that chromophore formation is only catalyzed in the presence of DNA sequences that incorporate binding sites for both zinc fingers.. AB - We ...
The gene encoding green fluorescent protein (GFP) has recently become an important visual marker of gene expression in eukaryotic organisms, as it is more sensitive than other reporter genes, requires no special cofactors for detection (7), and can be quantitated with a spectrofluorimeter (24). GFP has not been as widely applied to prokaryotic organisms because of a lack of constructs useful for diverse groups of bacteria, although GFP vectors are available for specialized bacterial systems (13, 24, 33, 41, 42). The wild-type gfpgene has been mutated to improve detection and expression of the fluorescent protein in prokaryotes (10, 18, 30), and both the wild-type and mutated forms have been used to construct less specialized bacterial GFP vectors.. A broad-host-range plasmid expressing the improved gfp(mut2) (10) gene from either a lac or annpt-2 promoter has been used successfully to tag gram-negative soil bacteria with GFP (27). Escherichia coli-Pseudomonas spp. shuttle vectors ...
TY - JOUR. T1 - Dynamics of insulin-stimulated translocation of GLUT4 in single living cells visualised using green fluorescent protein. AU - Dobson, SP. AU - Livingston, C. AU - Gould, GW. AU - Tavaré, JM. PY - 1996. Y1 - 1996. M3 - Article (Academic Journal). VL - 393. SP - 179. EP - 184. JO - FEBS Letters. JF - FEBS Letters. SN - 0014-5793. ER - ...
Sino Biologcial offers high quality green fluorescent protein GFPSpark® with features of bright green fluorescence, high pH-stability and fast maturation.
The recent emergence of an autofluorescent protein, the green fluorescent protein (GFP), has opened the door for the convenient use of intact living cells and organisms as experimental systems in fields ranging from cell biology to biomedicine. We present an overview of some of the major applications of GFP, namely its use in protein tagging and in monitoring gene expression as well as its potential in a variety of biological screens.. ...
As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards. The RCSB PDB also provides a variety of tools and resources. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.
UBC-GFP transgenic mice express green fluorescent protein directed by the human ubiqutin C promoter, and were discovered to have transgene insertion on chromosome 17 resulting in linkage to H-2|sup|b|/sup| MHC haplotype (see details below). Certain hematopoietic cell types display distinct expression levels of GFP, allowing identification of different cells types by FACS analysis. This strain is a useful tool for studying hematopoietic cell differentiation and in vivo leukocyte tracking. This transgene is also available on C57BL/6J as Stock No. |a href=|004353|/a|. |br /||br /||strong|In 2020, the UBC-GFP transgene was determined to have integrated at Chr17:29,435,589 - a noncoding region centromeric from the H-2 locus (~4.6 Mbp away from |em|H2-K1|/em|) - and is closely linked to H-2|sup|b|/sup| MHC haplotype. See Important Note for additional details|/strong|.
Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol...
Use of a dicistronic expression cassette encoding the green fluorescent protein for the screening and selection of cells expressing inducible gene products
A calculator mutagenesis technique has been used to characterize the structural effects associated with more than 46,000 amino acid variants simple and multiple green fluorescent proteins Aequorea ...
Green lights in the dark When someone first shows up in our lab, the prime goal I set up for him or her is to make green cells - I mean to introduce a Green Fluorescent Protein into a mammalian cell culture. In order to be able to perform this one has to know some basic molecular…
Shop a large selection of products and learn more about Thermo Scientific Lab Vision GFP (Green Fluorescent Protein) Ab-1, Mouse 100µL; 200µg/mL; Unlabeled; Purified
Photodamage of symbiotic algae exposed to thermal stress is involved in mass coral bleaching, a major cause of reef decline. Photoprotection is therefore a vital part of coral stress physiology. Corals produce a variety of green fluorescent protein (GFP)-like proteins from which some representatives screen the symbiotic algae from excess light. Different tissue concentrations of these GFP-like proteins distinguish colour morphs that are characteristic for many coral species. The question arises whether these pigmentation differences may diversify the niches that can be occupied by corals along the steep light gradient that structures coral reef communities. We assessed the implications of GFP-like protein expression in two color morphs of the symbiotic coral Hydnophora grandis, both associated with the same Symbiodinium sp. (subclade C40). The colour morphs of this species (high fluorescent, HF; and low fluorescent, LF), characterized by markedly different contents of a cyan fluorescent protein, were
In most previous papers concerning nisin quantification the authors emphasized the lowest detectable amount of nisin, which was given as the final assay concentration. However, this value seldom describes the most important numerical value giving true limits to the usefulness of the method in question, namely, the lowest detectable concentration of nisin in a food sample. Indeed, in some studies food material was spiked with amounts of nisin far from the linear dose-response range of the assay described, and the authors did not reveal the solvent into which the food extract was diluted prior to measurement (3, 5). Because of the sensitivity of immunological methods to interfering substances in a sample matrix, this kind of reporting makes it impossible for a reader to decide whether the assay in question is useful for his or her application.. At present, the most widely used quantification assay for nisin, the agar diffusion method, which was developed by Tramer and Fowler in 1964, is more ...
Green fluorescent protein (GFP), a 27 kDa protein derived from the jellyfish Aequorea victoria, emits green light (emission peak 509 nm) when excited by blue light (excitation peak 395 nm). GFP has become an invaluable tool in cell biology research, since its intrinsic fluorescence can be visualized in living cells. GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP has been widely used as a reporter for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. Other applications of GFP include assessment of protein-protein interactions through the yeast two hybrid system and measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. GFP is used to measure single cell metastasis and successful proliferation of stem cells.. ...
A complementary DNA for the Aequorea victoria green fluorescent protein (GFP) produces a fluorescent product when expressed in prokaryotic (Escherichia coli) or eukaryotic (Caenorhabditis elegans) cells. Because exogenous substrates and cofactors are not required for this fluorescence, GFP expression can be used to monitor gene expression and protein localization in living organisms. ...
Fig. 4 Functional assays show increased transformation potential and sensitivity to TNK2 inhibition.. (A) Total colony formation in mouse bone marrow colony formation assay. Mouse bone marrow cells were cotransduced to express PTPN11, PTPN11 E76K, TNK2, or empty vector controls. Cells were selected for GFP+ (green fluorescent protein-positive) and puromycin resistance and plated in a methylcellulose GM-CSF sensitivity colony formation assay. Colonies were counted at 14 days [GM-CSF] = 0.05 nM (0.71 ng/ml). ****P , 0.0001 by one-way ANOVA. (B) Total colony formation in mouse bone marrow colony formation assay in cells transduced with PTPN11, PTPN11 E76K, or PTPN11 G60R. Cells were sorted for GFP+. Cells were plated with increasing concentrations of dasatinib. ***P , 0.005 and ****P , 0.0005 by one-way ANOVA. (C) Total colony formation and percent total colony formation in mouse bone marrow colony formation assay. Mouse bone marrow cells were cotransduced to express PTPN11 E76K and TNK2 or TNK2 ...
Magnetic resonance imaging (MRI) of magnetically labeled stem cells has become a valuable tool in the understanding and evaluation of experimental stem cell-based therapies of degenerative central nervous system disorders. This comprehensive study assesses the impact of magnetic labeling of both human and rodent stem cell-containing populations on multiple biologic parameters as maintenance of stemness and oxidative stress levels. Cells were efficiently magnetically labeled with very small superparamagnetic iron oxide particles. Only under the condition of tailored labeling strategies can the impact of magnetic labeling on vitality, proliferation, pluripotency, and oxidative stress levels be minimized. In a rat model of Parkinson disease, magnetically labeled mouse embryonic stem cells were tracked by high-field MRI for 6 months. Significant interindividual differences concerning the spatial distribution of cells became evident. Histologically, transplanted green fluorescent protein-positive ...
Neuropeptide S (NPS) has been associated with a number of complex brain functions, including anxiety-like behaviors, arousal, sleep-wakefulness regulation, drug-seeking behaviors, and learning and memory. In order to better understand how NPS influences these functions in a neuronal network context, it is critical to identify transmitter systems that control NPS release and transmitters that are co-released with NPS. For this purpose, we generated several lines of transgenic mice that express enhanced green-fluorescent protein (EGFP) under control of the endogenous NPS precursor promoter. NPS/EGFP-transgenic mice show anatomically correct and overlapping expression of both NPS and EGFP. A total number of similar to 500 NPS/EGFP-positive neurons are present in the mouse brain, located in the pericoerulear region and the Kolliker-Fuse nucleus. NPS and transgene expression is first detectable around E14, indicating a potential role for NPS in brain development. EGFP-positive cells were harvested by ...
Funded by the NIH National Center for Advancing Translational Sciences through its Clinical and Translational Science Awards Program, grant number UL1TR002541 ...
Spinal cord injuries (SCI) are disastrous neuropathologies causing permanent disabilities. The availability of different strains of mice is valuable for studying the pathophysiological mechanisms involved in SCI. However, strain differences have a profound effect on spontaneous functional recovery after SCI. CX3CR1+/eGFP and Aldh1l1-EGFP mice that express green fluorescent protein in microglia/monocytes and astrocytes, respectively, are particularly useful to study glial reactivity. Whereas CX3CR1+/eGFP mice have C57BL/6 background, Aldh1l1-EGFP are in Swiss Webster background. We first assessed spontaneous functional recovery in CX3CR1+/eGFP and Aldh1l1-EGFP mice over 6 weeks after lateral spinal cord hemisection. Second, we carried out a longitudinal follow-up of lesion evolution using in vivo T2-weighted magnetic resonance imaging (MRI). Finally, we performed in-depth analysis of the spinal cord tissue using ex vivo T2-weighted MRI as well as detailed histology. We demonstrate that CX3CR1+/eGFP mice
TY - JOUR. T1 - Drug inhibition of Gly-Sar uptake and hPepT1 localization using hPepT1-GFP fusion protein. AU - Sun, Duxin. AU - Landowski, Christopher P.. AU - Chu, Xiaoyan. AU - Wallsten, Richard. AU - Komorowski, Thomas E.. AU - Fleisher, David. AU - Amidon, Gordon L.. PY - 2001/12/1. Y1 - 2001/12/1. N2 - An hPepT1-GFP fusion construct was made to study drug inhibition of dipeptide uptake and apical, basolateral, or subcellular hPepT1 localization. The hPepT1 stop codon was mutated by polymerase chain reaction and was subsequently cloned into the pEGFP-N1 vector. The hPepT1-GFP fusion construct was then transfected into Caco-2 and HeLa cells, and drug inhibition was studied by inhibiting 3H-Gly-Sar uptake. Western blot analysis was used to determine hPepT1-GFP expression levels and confocal microscopy was used to examine the localization. Both anti-hPepT1 antibody and anti-GFP antibody recognized a 120kd hPepT1-GFP fusion protein in the transfected cells. The 3H-Gly-Sar uptake in transfected ...
Green fluorescent protein (GFP), molecular model. The molecule has a cylindrical structure formed from beta sheets (ribbons). GFP is found in the Pacific jellyfish Aequorea victoria. It fluoresces green when blue light is shone on it. GFP is widely used as a research tool in biology and medicine. The gene coding for it can be tagged to the genes of other proteins or viruses to study their movements within cells. They can also be used to tag cancer cells to track their spread through the body. - Stock Image F006/9343
Purpose : The purpose of the current study is to determine in-depth functions of selected transcripts that are enriched in rod photoreceptors, in order to gain insights into intrinsic mechanisms underlying rod development and regeneration. Methods : We used a transgenic zebrafish line (XOPS:eGFP) in which rod photoreceptors express green fluorescent protein (GFP) as a model organism for this study. RNA-seq of FACS-sorted dissociated retinal cells was performed to identify differentially expressed (in GFP+ vs. GFP- cells) transcripts with FDR , 0.01. Selected rod-specific genes were prioritized for further qRT-PCR and in situ hybridization studies at different life stages of the zebrafish, and for qRT-PCR studies of rods that regenerated after widespread chemical lesioning of the retina. The rxrγa gene was of particular interest as its transcript was enriched in rods, while previous studies in mouse indicated roles in cone determination. Therefore, a new rxrγa mutant line was created by ...
TY - JOUR. T1 - Probing intra-molecular mechanics of single circularly permuted green fluorescent protein with atomic force microscopy. AU - Wang, Tong. AU - Nakajima, Ken. AU - Miyawaki, Atsushi. AU - Hara, Masahiko. PY - 2005/11/1. Y1 - 2005/11/1. N2 - We investigated the mechanical unfolding of single circularly permuted green fluorescent protein (cpGFP) with atomic force microscopy (AFM). The molecule was stretched from its N- and C-termini by an external force causing an elongation of the polypeptide chain up to its full length. The features of the force-extension (F-E) curves were found to depend on the stretching speed. At fast speeds, we detected one peak in the F-E curves before final rupture of the extended molecule, which we interpreted as the unfolding of two terminal halves within cpGFP. We observed several more force peaks in a sawtooth pattern at much slower speeds, and explained the appearance of such force peaks as cooperative unfolding of the hidden sub-structures inside each ...
Hi, brief question. does anti-GFP from invitrogen can recognize EGFP from clontech? I am currently trying to detect EGFP on western blot by using anti-GFP from invitrogen but I cannot detect it. Also anybody working with anti-GFP from invitrogen and having any comment? should I use anti-GFP from other company? mUsClemUsClemUsClemUsClemUsCle Keitaro YAMANOUCHI D. V. M., Ph. D. Muscle Biology Group Nutritional Science University of Arizona 601 Shantz Building Tucson AZ 85721-0038 Tel: 520-621-3829 Fax: 520-621-1396 e-mail: keitaro at mUsClemUsClemUsClemUsClemUsCle ...
In the present paper, we introduce a transgenic mouse line whose sperm express green fluorescent protein (GFP) in their acrosome and red fluorescent protein (RFP) in their mitochondria [B6D2F1- Tg(CAG/su9-DsRed2, Acr3-EGFP)RBGS002Osb]. The dual fluorescent sperm showed normal fertilizing ability in …
Simple and easy to engineer metal-sensing molecules that are capable of differentiating metal ions and producing metal-specific signals are highly desirable. Metal ions affect the thermal stability of proteins by increasing or decreasing their resistance to unfolding. This work illustrates a new strategy for designing bivalent fluorescent fusion proteins capable of differentiating metal ions in solution through their distinct effects on a proteins thermal stability. A new dual purpose metal sensor was developed consisting of biotin protein ligase (BirA) from B. pseudomallei (Bp) fused to green fluorescent protein (GFP). When coupled with differential scanning fluorimetry of GFP-tagged proteins (DSF-GTP) for signal-transduction detection, Bp BirA-GFP yields distinct protein unfolding signatures with Zn(II) and Cu(II) ions in aqueous solutions. The limit of detection of the system is ∼1 μM for both metal species. The system can be used in a variety of high-throughput assay formats including ...
Light micrograph (LM Fluorescence) of a transformed bacteria colony (Escherichia coli) containing a jellyfish gene for GFP (green fluorescent protein). GFP is a 238 amino acid protein found in the Pacific jellyfish (Aequorea victoria). It exhibits a bright green fluorescence (green bioluminescence) when exposed to ultraviolet blue light. GFP is widely used as a research tool in biology and medicine. The gene coding for GFP can be tagged to the genes of other proteins or viruses to study their movements within cells. GFP can also be used to tagged cancer cells to track their spread through the body. The purpose of both bioluminescence and GFP fluorescence in jellyfish is unknown. Magnification: x20 when shortest axis printed at 25 millimetres. - Stock Image C032/2759
Over the past several years, we have differentiated green fluorescent protein-expressing human embryonic stem cells (GFP-hESC) into mesenchymal stem cell-like cells (eMSCs). These eMSCs expressed markers (CD-29, CD-44, CD-73, CD-105, CD-166 and Nestin, but not CD-34, C45, CD-106, SSEA-4 or Oct-4) which are consistent with a mesenchymal stem cell state. We have transplanted eMSCs into the femoral veins of SHRs following MCAO. The expression of GFP allows us to follow the migration and further differentiation of the eMSCs in the ischemic brain. We observed migration of the injected eMSCs to the infarction region (Fig. 1) and their subsequent differentiation into neurons (which expressed β-tubulin III, MAP2, HuC and neurofilament) (see Fig. 2) and vascular endothelial cells (which expressed vWF; see Fig. 3). The grafted cells do not appear to differentiate into astrocytes.. ...
Over the past several years, we have differentiated green fluorescent protein-expressing human embryonic stem cells (GFP-hESC) into mesenchymal stem cell-like cells (eMSCs). These eMSCs expressed markers (CD-29, CD-44, CD-73, CD-105, CD-166 and Nestin, but not CD-34, C45, CD-106, SSEA-4 or Oct-4) which are consistent with a mesenchymal stem cell state. We have transplanted eMSCs into the femoral veins of SHRs following MCAO. The expression of GFP allows us to follow the migration and further differentiation of the eMSCs in the ischemic brain. We observed migration of the injected eMSCs to the infarction region (Fig. 1) and their subsequent differentiation into neurons (which expressed β-tubulin III, MAP2, HuC and neurofilament) (see Fig. 2) and vascular endothelial cells (which expressed vWF; see Fig. 3). The grafted cells do not appear to differentiate into astrocytes.. ...
Abstract: Background: Sterile α motif and HD domain-containing protein-1 (SAMHD1) inhibits HIV-1 reverse transcription by decreasing the pool of intracellular deoxynucleotides. SAMHD1 is controlled by cyclin-dependent kinase (CDK)-mediated phosphorylation. However, the exact mechanism of SAMHD1 regulation in primary cells is unclear. We explore the effect of palbociclib, a CDK6 inhibitor, in HIV-1 replication.. Methods: Human primary monocytes were differentiated into macrophages with monocyte-colony stimulating factor and CD4+ T lymphocytes stimulated with phytohaemagglutinin (PHA)/interleukin-2. Cells were treated with palbociclib and then infected with a Green fluorescent protein-expressing HIV-1 or R5 HIV-1 BaL. Viral DNA was measured by quantitative PCR and infection assessed by flow cytometry. Deoxynucleotide triphosphate (dNTP) content was determined using a polymerase-based method.. Results: Pan-CDK inhibitors AT7519, roscovitine and purvalanol A reduced SAMHD1 phosphorylation. HIV-1 ...
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
mKate2 is the next generation of monomeric far-red fluorescent protein TagFP635 (mKate) [Shcherbo et al., 2007; Shcherbo et al., 2009]. Possessing fluorescence with excitation/emission maxima at 588 and 633 nm, mKate2 is almost 3-fold brighter than TagFP635 and is 10-fold brighter than mPlum at the physiological pH = 7.5. Within the optical window optimal for light penetration in living tissues, calculated brightness of mKate2 is at least 2-fold higher compared to any monomeric fluorescent protein reported to date. mKate2 is characterized by complete and fast chromophore maturation at 37°C with maturation half-time ,20 min (versus 40 min for mCherry). It is more photostable under both widefield and confocal illumination than other monomeric far-red proteins, including TagFP635, mRaspberry and mPlum. The high brightness, far-red emission spectrum, excellent pH resistance and photostability, coupled with low toxicity demonstrated in transgenic Xenopus laevis embryos, make mKate2 a superior ...
Need antibody products for research? Find and compare multiple sources of anti-Green Fluorescent Protein (GFP) antibody using the Linscotts Directory search engine. Monoclonal, polyclonal, and recombinant antibodies. Select applications, conjugates, hosts, and reactivity. Get complete supplier details here.
Need antibody products for research? Find and compare multiple sources of anti-Green Fluorescent Protein (GFP) antibody using the Linscotts Directory search engine. Monoclonal, polyclonal, and recombinant antibodies. Select applications, conjugates, hosts, and reactivity. Get complete supplier details here.
Green-to-red photoconversion is a reaction that occurs in a limited number of fluorescent proteins and that is currently mechanistically debated. In this contribution, we report on our investigation of the photoconvertible fluorescent protein Dendra2 by employing a combination of pump-probe, up-conversion and single photon timing spectroscopic techniques. Our findings indicate that upon excitation of the neutral green state an excited state proton transfer proceeds with a time constant of 3.4 ps between the neutral green and the anionic green states. In concentrated solution we detected resonance energy transfer (25 ps time constant) between green and red monomers. The time-resolved emission spectra suggest also the formation of a super-red species, first observed for DsRed (a red fluorescent protein from the corallimorph species Discosoma) and consistent with peculiar structural details present in both proteins.
Transgenic techniques have rapidly evolved in recent years. However, the efficiency of these techniques to produce viable offspring is still disappointingly low. The purpose of this study was to assess in vitro development, transgene expression, and integration following pronuclear or cytoplasmic microinjection of condensed or linear green fluorescent protein DNA into murine embryos using electroporation. In experiment 1, the effect of embryo orientation (group or linear) within the electroporation chamber on development was evaluated using zygotes which received one pulse duration (10 msec), and one of two voltages (250 or 400 V). Zygotes that received 400 V had the lowest development score (Group, 2.06 ? 0.12; Linear, 1.97 ? 0.13), irrespective of orientation. Embryos that received 250 V had the highest development of the voltage treated groups (Group 3.42 ? 0.12; Linear 3.32 ? 0.12), irrespective of orientation, and development was lower than the control embryos (Control 4.28 ? 0.12; Mannitol ...
Staphylococcus aureus AgrC is an important component of the agr quorum-sensing system. AgrC is a membrane-embedded histidine kinase that is thought to act as a sensor for the recognition of environmental signals and the transduction of signals into the cytoplasm. However, the difficulty of expressing and purifying functional membrane proteins has drastically hindered in-depth understanding of the molecular structures and physiological functions of these proteins. Here, we describe the high-yield expression and purification of AgrC, and analyze its kinase activity. A C-terminal green fluorescent protein (GFP) fusion to AgrC served as a reporter for monitoring protein expression levels in real time. Protein expression levels were analyzed by the microscopic assessment of the whole-cell fluorescence. The expressed AgrC-GFP protein with a C-terminal His-tagged was purified using immobilized metal affinity chromatography (IMAC) and size exclusion chromatography (SEC) at yields of ≥10 mg/L, following
Bone marrow harbors cells that have the capacity to differentiate into cells of nonhematopoietic tissues of neuronal, endothelial, epithelial, and muscular phenotype. Here we demonstrate that bone marrow-derived cells populate pancreatic islets of Langerhans. Bone marrow cells from male mice that express, using a CRE-LoxP system, an enhanced green fluorescent protein (EGFP) if the insulin gene is actively transcribed were transplanted into lethally irradiated recipient female mice. Four to six weeks after transplantation, recipient mice revealed Y chromosome and EGFP double-positive cells in their pancreatic islets. Neither bone marrow cells nor circulating peripheral blood nucleated cells of donor or recipient mice had any detectable EGFP. EGFP-positive cells purified from islets express insulin, glucose transporter 2 (GLUT2), and transcription factors typically found in pancreatic β cells. Furthermore, in vitro these bone marrow-derived cells exhibit - as do pancreatic β cells - ...
P. Didier, L. Guidoni, and E. Weiss, Ultrafast Excited-state Dynamics of Genetic Fusions: The Green Fluorescent Protein as a Folding Reporter, in Conference on Lasers and Electro-Optics/Quantum Electronics and Laser Science Conference and Photonic Applications Systems Technologies, Technical Digest (CD) (Optical Society of America, 2006), paper QMH1 ...
A reporter gene assay using the human placental secreted alkaline phosphatase (SEAP) or the green fluorescent protein for G proteiencoupled receptors
Neurons and glia are derived from a common set of precursor stem cells. Morrow et al. used a cortical slice assay to examine the signals required to specify neuronal versus glial cell fate and differentiation. Dissociated embryonic neural stem cells from a green fluorescent protein (GFP)-expressing strain of mice were plated with cortical slices from mice of different ages (embryonic through postnatal). Cell fate and differentiation were monitored by changes in the morphology and expression of marker proteins for neurons and glia. Incubation of neural stem cells with embryonic cortical slices led to the development of mostly neuronal cells and incubation with postnatal cortical slices produced mostly glial cells, suggesting that different factors are produced at different stages of development and that the stem cells are poised to respond to either set of signals. Analysis of clones of the GFP-expressing cells showed that the signals were predominantly instructive and not trophic signals. The ...
We combine Fluorescence Recovery After Photobleaching (FRAP) experiments with mathematical modelling to study the dynamics inside the nucleus of both the TGF-β-sensitive transcriptional regulator Smad2, and Green-Fluorescent Protein (GFP). We show how combining modelling with bleaching strips of different areas allows a rigorous test of whether or not a protein is moving via diffusion as a single species. As noted recently by others, it is important to consider diffusion during the bleaching process. Neglecting it can cause serious error. Also, it is possible to use the bleaching process itself to provide an extra consistency test to the models predicting the recovery. With our method we show that the dynamics of GFP are consistent with it diffusing as a single species in a uniform environment in which flow is negligible. In contrast, the dynamics of the intracellular signal transducer Smad2 are never consistent with it moving as a single species via simple diffusion in a homogeneous ...
Mono- and Stereopictres of 5.0 Angstrom coordination sphere of Copper atom in PDB 1kyr: Crystal Structure of A Cu-Bound Green Fluorescent Protein Zn Biosensor
Fingerprint Dive into the research topics of Sensitivity of the yellow variant of green fluorescent protein to halides and nitrate [2]. Together they form a unique fingerprint. ...
The advent of jellyfish green fluorescent protein and its spectral variants, together with promising new fluorescent proteins from other classes of the Cnidarian phylum (coral and anemones), has greatly enhanced and promises to further boost the detection and localization of proteins in cell biology. It has been less widely appreciated that highly sensitive methods have also recently been developed for detecting the movement and localization in living cells of the very molecules that precede proteins in the gene expression pathway, i.e. RNAs. These approaches include the microinjection of fluorescent RNAs into living cells, the in vivo hybridization of fluorescent oligonucleotides to endogenous RNAs and the expression in cells of fluorescent RNA-binding proteins. This new field of fluorescent RNA cytochemistry is summarized in this article, with emphasis on the biological insights it has already provided. These new techniques are likely to soon collaborate with other emerging approaches to advance the
Sandia National Laboratories researchers are drawing inspiration from neurons in the brain, such as these green fluorescent protein-labeled neurons in a 1f52c read all about
Our researchers have developed a new technology that allows for the study of protein structure/folding/functions in the living cells. This new technology involves expressing a protein using bacteria and labeling the protein with probes. Researchers can then modify the recombinant protein with a special reagent that allows for the transfer of the modified protein into the living mammalian cells. This will also allow for the study of protein traffic in the cells. Furthermore, this new technology permits high-resolution structural biology techniques to be combined with cell biology techniques and provides a foundation for future applications of protein transduction technology, atomic resolution cell biology, and protein drug therapy to treat human disease. ...
Stem cells have great potential value for treating a number of diseases and conditions, including diabetes, Parkinsons, and spinal cord injuries. Applying stem cells for therapeutic purposes will require an in-depth understanding of their biology, not only of the genes they express, but also the functions of the proteins encoded by the genes. The goal of the project presented in this thesis was to develop a method for high-throughput analyses of protein localization in mouse stem cells. Localization information can provide insight into the functions and biological roles of proteins. ,br /,,br /, One means of studying protein localization involves creating proteins with a green fluorescent protein (GFP) reporter gene and analyzing their localization using fluorescence microscopy. The research outlined in this thesis focused on developing a system to create a large number of GFP-tagged proteins by constructing a cDNA?GFP fusion library. This involved exploring methods for optimizing cDNA ...
The effects of embryonic and larval ethanol exposure on brain development were visually monitored using transgenic zebrafish expressing cell-specific green fluorescent protein (GFP) marker genes. Specific subsets of GFP-expressing neurons were highly sensitive to ethanol exposure, but only during defined developmental windows. In the med12 mutant, which affects the Mediator co-activator complex component Med12, exposure to lower concentrations of ethanol was sufficient to reduce GFP expression in transgenic embryos. In transgenic embryos and larva containing GFP driven by an oxytocin-like (oxtl) promoter, ethanol exposure dramatically up-regulated GFP expression in a small group of hindbrain neurons, while having no effect on expression in the neuroendocrine preoptic area.. ...
A special protein found in jellyfish called green fluorescent protein (or GFP) glows green when a particular wavelength of light is shone on it. By following the green glow you know where GFP is. Can this help locate other proteins? GFP is expressed from a GFP gene and, like all genes, the GFP gene is made up of A, T, C and G subunits. This is important; it means that a gene from a person (e.g. the gene we found mutated in the Parkinsons family) can be attached to the jellyfish GFP gene to form a hybrid gene and therefore a hybrid protein: one half human and the other half jellyfish. Therefore, wherever the human protein goes the GFP protein goes too; when an expression plasmid containing the GFP hybrid gene is introduced into cells a particular part of the cell will glow green, demonstrating the human protein does its job there. ...
These tools will help advance any experiments utilizing Green Fluorescent Protein or Red Fluorescent Protein fusion constructs.. ...
Kicking off the evening was John Lee, from the University of Georgia, with his ambitiously titled talk Bioluminescence: The First 3000 Years. After a historical introduction to the long running observation of bioluminescence, via the discovery in 1672 that oxygen was necessary for bacterial luminescence, John told us how it was determined that bioluminescence is an enzyme mediated chemical reaction involving luciferase and luciferine. In the modern age of biochemistry it was determined that ATP is the substrate in this reaction. Following the elucidation of the structure of firefly luciferase in 1959, modern techniques (i.e. picosecond dynamic fluorescence spectroscopy and NMR) have allowed researchers to uncover the enzymes and processes involved in bioluminescence. One of the most important of these enzymes Green-fluorescent protein (GFP) was discovered in jellyfish by Shimomura (who evidently has a lab at his house!) and led to his Nobel prize in 2008. Due to GFPs widespread use in ...
Executive Home on Green Lake - 100 ft Sandy, Walkout Shorefront. This Executive Lake Home rests on Green Lake in Central Minnesota. Premier Green Lake is a...
Roger Tsien on fluorescent proteins Excitation and emission spectra for various fluorescent proteins Green Fluorescent Protein ... The green fluorescent protein (GFP) is a protein that exhibits bright green fluorescence when exposed to light in the blue to ... Green Fluorescent Protein on FPbase, a fluorescent protein database Overview of all the structural information available in the ... Protein methods, Recombinant proteins, Cell imaging, Protein imaging, Fluorescent proteins, Bioluminescence, Cnidarian proteins ...
... (YFP) is a genetic mutant of green fluorescent protein (GFP) originally derived from the jellyfish ... "Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin". Nature. 388 (6645): 882-7. doi:10.1038/ ... v t e (Articles with short description, Short description matches Wikidata, Fluorescent proteins, All stub articles, Protein ... "Crystal Structure of the Aequorea victoria Green Fluorescent Protein". Science. 273 (5280): 1392-1395. doi:10.1126/science. ...
A fluorescent green protein derived from this screen was serendipitously discovered to have sensitivity to ultraviolet light-- ... Photoactivatable fluorescent proteins (PAFPs) is a type of fluorescent protein that exhibit fluorescence that can be modified ... a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion". Proceedings of the National Academy of ... "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein". Proceedings of the National ...
FbFPs absorb blue light and emit light in the cyan-green spectral range. LOV-domains are a sub-class of PAS domains and were ... Recombinant proteins, Protein imaging, Protein methods, Cell imaging, Fluorescent proteins, Bioluminescence). ... A FMN-binding fluorescent protein (FbFP), also known as a LOV-based fluorescent protein, is a small, oxygen-independent ... Jung G, Brockhinke A, Gensch T, Hötzer B, Schwedler S, Veettil SK (2012). Fluorescent Proteins I. From Understanding to Design ...
... "the green fluorescent protein: discovery, expression and development." The multicolored fluorescent proteins developed in ... Typically, the gene coding for a protein of interest is fused with the gene for a fluorescent protein, which causes the protein ... "Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin". Nature. 388 (6645): 882-7. Bibcode: ... "Green Fluorescent Protein". The Golden Goose Award. Retrieved May 27, 2015. "Golden Plate Awardees of the American Academy of ...
An alternative approach is using recombinant proteins containing fluorescent protein domains, e.g., green fluorescent protein ( ... Chalfie M (October 1995). "Green fluorescent protein". Photochemistry and Photobiology. 62 (4): 651-656. doi:10.1111/j.1751- ... Likewise, an antigen can also be conjugated to the antibody with a fluorescent probe in a technique called fluorescent antigen ... Proteins in the supernatant or on the outside of the cell membrane can be bound by the antibodies; this allows for living cells ...
"Green Fluorescent Protein". The Golden Goose Award. Retrieved 2015-05-27. Wikimedia Commons has media related to Martin Chalfie ... 1994). "Green fluorescent protein as a marker for gene expression". Science. 263 (5148): 802-805. Bibcode:1994Sci...263..802C. ... He traces his work on green fluorescent protein to a 1988 seminar from Paul Brehm about bioluminescent organisms, which led to ... She gave him permission to cite her unpublished research in his seminal Science paper "Green Fluorescent Protein as a Marker ...
In 1962, their work culminated in the discovery of the proteins aequorin and green fluorescent protein (GFP) in A. victoria; ... "Green Fluorescent Protein". The Golden Goose Award. Archived from the original on 2015-09-09. Retrieved 2015-05-27. " ... He was awarded the Nobel Prize in Chemistry in 2008 for the discovery and development of green fluorescent protein (GFP) with ... Osamu Shimomura on including the Nobel lecture Discovery of Green Fluorescent Protein, GFP (All articles with ...
Green fluorescent protein is a naturally occurring fluorescent protein from the jellyfish Aequorea victoria that is widely used ... YFP or yellow fluorescent protein, BFP or blue fluorescent protein, and CFP or cyan fluorescent protein are examples of GFP ... fluorescent proteins or FPs were introduced. Green fluorescent protein or GFP was discovered by Osamu Shimomura in the 1960s ... Alterations of fluorescent proteins would lead to loss of fluorescent properties. Protein labeling use a short tag to minimize ...
... the green fluorescent protein, which gives a green glow if cells produce this type of protein Like most other circular plasmids ... "Green Fluorescent Protein History".{{cite web}}: CS1 maint: url-status (link) "The Nobel Prize in Chemistry 2008 ... The plasmid contains several reporter genes, most notably the green fluorescent protein (GFP) and the ampicillin resistance ... The pGLO plasmid was made famous by researchers in France who used it to produce a green fluorescent rabbit named Alba. Other ...
Tsien, Roger Y. (1998-01-01). "The Green Fluorescent Protein". Annual Review of Biochemistry. 67 (1): 509-544. doi:10.1146/ ... In contrast to fluorescent proteins which form their chromophore through posttranslational modifications of the polypeptide ... Biliverdin (latin for green bile) is a green tetrapyrrolic bile pigment, and is a product of heme catabolism. It is the pigment ... 2011). "Bright and stable near infra-red fluorescent protein for in vivo imaging". Nat Biotechnol. 29 (8): 757-761. doi:10.1038 ...
The full text Archived 2011-09-30 at the Wayback Machine Tsien, R. (1998). The green fluorescent protein. Annual Review of ... Aequoreids include Aequorea victoria, the organism from which the green fluorescent protein gene was isolated. Only the polyp ...
The aptamer was designed to be an RNA mimic of green fluorescent protein (GFP); similar to GFP for proteins, Spinach can be ... Paige, J. S.; Wu, K. Y.; Jaffrey, S. R. (2011). "RNA Mimics of Green Fluorescent Protein". Science. 333 (6042): 642-646. ... As the fluorophore of GFP and its derivatives are covalently bound to/a part of the protein, free exchange cannot happen and ... Spinach has also been adapted for sensing proteins or molecules in vivo. An adapted structure, which includes two binding sites ...
Elowitz, M. B.; Surette, M. G.; Wolf, P. E.; Stock, J.; Leibler, S. (1997). "Photoactivation turns green fluorescent protein ...
Paige JS, Wu KY, Jaffrey SR (July 2011). "RNA mimics of green fluorescent protein". Science. 333 (6042): 642-6. Bibcode:2011Sci ... fluorescent labeling of proteins and cells, and selective enzyme inhibition. Aptamer - Oligonucleotide or peptide molecules ... For example, in the case of negatively charged small molecules and proteins, high salt buffers are used for charge screening to ... Alternatively, if the desired aptamer function is in vivo protein or whole cell binding for potential therapeutic or diagnostic ...
Known for discovering green fluorescent protein. Chinary Ung, Music. Grawemeyer Award winning composer. Harold Urey, Chemistry ... How a fluorescent jellyfish - and federal dollars - helped fight AIDS, The Washington Post, Retrieved 14 September 2012. UCSD ...
Viral Applications of Green Fluorescent Protein. Methods in Molecular Biology (Clifton, N.J.). Vol. 515. pp. 165-175. doi: ... of the first recombinant HIV virus that enabled the visualization of HIV infected cells by expressing green fluorescent protein ... Brown also explored the sec-dependent protein export pathway in mycobacterium, the main protein export pathway into the ... Brown also used this fluorescent tool to discover that HLA-A2 is down-regulated in HIV infected macrophages. Further, Brown was ...
"Green fluorescent protein takes Nobel prize". Lewis Brindley. Retrieved 31 May 2015. Alberts B, Johnson A, Lewis J, Raff M, ... Mattingly CJ, McLachlan JA, Toscano WA (August 2001). "Green fluorescent protein (GFP) as a marker of aryl hydrocarbon receptor ... Scientists have genetically engineered several organisms, including some mammals, to include green fluorescent protein (GFP), ... The GloFish is a brand of genetically modified fluorescent zebrafish with bright red, green, and orange fluorescent color. It ...
... s naturally express green fluorescent proteins (GFP) inside their oral tentacles and near the eye spot. Depending on ... The fluorescent proteins from lancelets have been adapted for use in molecular biology and microscopy. The yellow fluorescent ... "Endogenous Green Fluorescent Protein (GFP) in Amphioxus". The Biological Bulletin. 213 (2): 95-100. doi:10.2307/25066625. ISSN ... "A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum". Nature Methods (published 24 March 2013 ...
The first fluorescent protein to be discovered, green fluorescent protein (GFP), has been adapted to identify and develop ... DsRed on FPBase (Protein articles without symbol, Protein pages needing a picture, Fluorescent proteins). ... Variants such as yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP) were discovered in Anthozoa. Issues with ... Red fluorescent protein (RFP) is a fluorophore that fluoresces red-orange when excited. Several variants have been developed ...
In the animal, the protein occurs together with the green fluorescent protein to produce green light by resonant energy ... It was also noted during the extraction the animal creates green light due to the presence of the green fluorescent protein, ... "Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin". Nature. 388 (6645): 882-887. Bibcode: ... Shimomura O (2005). "The discovery of aequorin and green fluorescent protein". J Microsc. 217 (Pt 1): 1-15. doi:10.1111/j.0022- ...
Inserting one protein as a domain into another protein can be useful. For instance, inserting calmodulin into green fluorescent ... "Quantitative in vivo solubility and reconstitution of truncated circular permutants of green fluorescent protein". Protein ... Regardless of which protein comes first, this fusion protein may show similar function. Thus, if a fusion between two proteins ... Jung J, Lee B (September 2001). "Circularly permuted proteins in the protein structure database". Protein Science. 10 (9): 1881 ...
There are three types of screening commonly used: Green fluorescent protein makes cells glow green under UV light. A ... "Green fluorescent protein as a marker for gene expression." Science 263.5148 (1994): 802-805. Biomarker Recombinant DNA (Genes) ...
Cormack, B. P.; Valdivia, R. H.; Falkow, S. (1996). "FACS-optimized mutants of the green fluorescent protein (GFP)". Gene. 173 ...
... phycoerythrin or green fluorescent protein). Alternatively, specific or general proteins, nucleic acids, lipids or small ... Several fluorescent protein exist in nature[citation needed], but the most important one as a research tool is Green ... Chalfie, M; Tu, Y; Euskirchen, G; Ward, WW; Prasher, DC (1994). "Green fluorescent protein as a marker for gene expression". ... Other proteins are fluorescent but require a fluorophore cofactor, and hence can only be used in vitro; these are often found ...
It also exhibits proteins similar to green fluorescent protein. Norzoanthamine (2018). Zoanthus Lamarck, 1801. Accessed through ...
Rizzuto R, Brini M, Pizzo P, Murgia M, Pozzan T (June 1995). "Chimeric green fluorescent protein as a tool for visualizing ... Kneen M, Farinas J, Li Y, Verkman AS (March 1998). "Green fluorescent protein as a noninvasive intracellular pH indicator". ... For measuring pH inside of organelles, a technique utilizing pH-sensitive green fluorescent proteins (GFPs) may be used. ... "pH difference across the outer mitochondrial membrane measured with a green fluorescent protein mutant". Biochemical and ...
"Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin". Nature. 388 (6645): 882-887. Bibcode: ... Fluorescent tagging uses a gene encoding a fluorescent protein that is inserted into the coding frame of the protein to be ... "Green fluorescent protein as a marker for gene expression." Trends in Genetics 10.5 (1994): 151. Ng, T. (1999). "Imaging ... The field of live single-cell imaging began with work demonstrating that green fluorescent protein (GFP), found in the ...
"Novel fluorescent protein from Hydnophora rigida possesses green emission". Biochemical and Biophysical Research Communications ... Their color is naturally green and brown, or sometimes cream. They can also become fluorescent green and cyano-red emission. H ... Bokhari, H.; Smith, C.; Veerendra, K.; Sivaraman, J.; Sikaroodi, M.; Gillevet, P. (June 2010). "Novel fluorescent protein from ... H. rigida has thin branches that are cream or green without the encrusting bases. The horn coral also has a green fluorescence ...
While the use of fluorescent proteins was once limited to the green fluorescent protein (GFP), in recent years many other ... EosFP is a photoactivatable green to red fluorescent protein. Its green fluorescence (516 nm) switches to red (581 nm) upon UV ... "New Fluorescent Protein Permanently Marks Neurons that Fire". Retrieved 2017-12-01. (Fluorescent proteins, ... "Structural basis for photo-induced protein cleavage and green-to-red conversion of fluorescent protein EosFP". Proceedings of ...
... green fluorescent protein, lipophylic dyes or radioactively tagged amino acids) into the brain. These molecules are absorbed ... There is also a group of tracers that consist of protein products that can be taken up by the cell and transported across the ... virus or protein can be locally injected, after which it is allowed to be transported anterogradely. Viral tracers can cross ...
Originally, a green fluorescent protein, mAG, was fused to hGem(1/110) and an orange fluorescent protein (mKO2) was fused to ... but are not functional proteins. The green fluorescent protein is made during the S, G2, or M phase and degraded during the G0 ... Rodriguez EA, Tran GN, Gross LA, Crisp JL, Shu X, Lin JY, Tsien RY (September 2016). "A far-red fluorescent protein evolved ... A far-red and near-infrared FUCCI was developed using a cyanobacteria-derived fluorescent protein (smURFP) and a ...
... some of the blue light released by aequorin in contact with calcium ions is absorbed by a green fluorescent protein, which in ... Nordgren, I. K.; Tavassoli, A. (2014). "A bidirectional fluorescent two-hybrid system for monitoring protein-protein ... Tsien won the 2008 Nobel Prize in Chemistry for their 1961 discovery and development of green fluorescent protein as a tool for ... In Vivo luminescence cell and animal imaging uses dyes and fluorescent proteins as chromophores. The characteristics of each ...
... is closely associated with a luciferin-binding protein as well as a green fluorescent protein (GFP). Calcium triggers release ... for many of the same applications as fluorescent proteins. However, unlike fluorescent proteins, luciferases do not require an ... Massoud TF, Paulmurugan R, De A, Ray P, Gambhir SS (Feb 2007). "Reporter gene imaging of protein-protein interactions in living ... Luciferase is a heat-sensitive protein that is used in studies on protein denaturation, testing the protective capacities of ...
BSE prions are misfolded forms of the particular brain protein called prion protein. When this protein is misfolded, the normal ... Wells GA, Hawkins SA, Green RB, Austin AR, Dexter I, Spencer YI, et al. (January 1998). "Preliminary observations on the ... After amplifying and then concentrating any PrPSc, the samples are labelled with a fluorescent dye using an antibody for ... BSE is thought to be due to an infection by a misfolded protein, known as a prion. Cattle are believed to have been infected by ...
... usually fused to a reporter such as green fluorescent protein. Initial studies determined that amino acids 9-52 from the rat ... Johnson HW, Schell MJ (2009). "Neuronal IP3 3-kinase is an F-actin-bundling protein: role in dendritic targeting and regulation ... It consists of a portion of the amino terminal actin binding region of the rat protein ITPKA, ...
Apple-green birefringence of Congo red stained preparations under polarized light is indicative of the presence of amyloid ... The manufacturer reports that fluorescent light through a Congo Blue filter gives the appearance of black light. Congo Blue ... allowing effector proteins to pass through and alter the host cell's biochemistry. The dye can also be used in flow cytometry ... In confocal microscopy, Congo red can be used as a stable fluorescent stain. Klaus Hunger, Peter Mischke, Wolfgang Rieper, ...
It is widely used as a fluorescent tracer for many applications. The color of its aqueous solutions is green by reflection and ... FITC reacts with the amine groups of many biologically relevant compounds including intracellular proteins to form a thiourea ... The fluorescein that has been taken up into the plant can be visualized under a fluorescent microscope. Chemical derivatives of ... In plant science, fluorescein, and other fluorescent dyes, have been used to monitor and study plant vasculature, particularly ...
"Genetically Encoded Indicators of Cellular Calcium Dynamics Based on Troponin C and Green Fluorescent Protein". Journal of ... that relies on a fusion protein that combines a synaptic vesicle membrane protein and a pH sensitive fluorescent protein. Upon ... "Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins". Nature. 394 (6689): 192-195. ... Genetically encoded voltage sensitive fluorescent proteins have also been developed. Calcium imaging relies on dyes or ...
While the SYBR Green dye emits its fluorescent signal simply by binding to the double-stranded DNA in solution, the TaqMan ... engineered a mutation of a protein suspected to participate in the regulation of Gal genes. This mutation was hypothesized to ... SYBR Green When the SYBR Green binds to the double-stranded DNA of the PCR products, it will emit light upon excitation. The ... It is also the preferred method of analysis when using DNA binding dyes such as SYBR Green since the elimination of primer- ...
... awarded the Nobel Prize in Chemistry in 2008 for his discovery and development of green fluorescent protein with two American ...
Some of these proteins are linked to the exterior of the cell membrane. An example of this is the CD59 protein, which ... A natural lipid bilayer is not fluorescent, so at least one fluorescent dye needs to be attached to some of the molecules in ... Coones, R. T.; Green, R. J.; Frazier, R. A. (2021). "Investigating lipid headgroup composition within epithelial membranes: a ... The most common class of this type of protein is the G protein-coupled receptor (GPCR). GPCRs are responsible for much of the ...
"Phase I clinical trial of a genetically modified and oncolytic vaccinia virus GL-ONC1 with green fluorescent protein imaging ... angiogenesis modifying proteins, radiosensitzing proteins, prodrug converting enzymes, biomarkers and immunomodulatory proteins ... of Genelux includes numerous genetically modified vaccinia virus strains with transgenes encoding for reporter proteins for ...
The matrix protein (M1) and membrane protein (M2) share a segment, as do the non-structural protein (NS1) and the nuclear ... Mordini E, Green M, eds. (2013). Internet-Based Intelligence in Public Health Emergencies: Early Detection and Response in ... Direct fluorescent or immunofluorescent antibody (DFA/IFA) tests involve staining respiratory epithelial cells in samples with ... disrupting internal protein-protein interactions to release RNPs into the host cell's cytosol. The M1 protein shell surrounding ...
... binding protein], PACT (protein activator of the interferon-induced protein kinase), the SMN complex, fragile X mental ... Using a lineage tracing approach followed by Fluorescent-activated cell sorting, miRNA profiling of the FoxD1-derived cells not ... Djuranovic S, Nahvi A, Green R (April 2012). "miRNA-mediated gene silencing by translational repression followed by mRNA ... HMGA proteins (HMGA1a, HMGA1b and HMGA2) are implicated in cancer, and expression of these proteins is regulated by microRNAs. ...
Rawstron AC, Green MJ, Kuzmicki A, Kennedy B, Fenton JA, Evans PA, et al. (July 2002). "Monoclonal B lymphocytes with the ... Smudge cells are due to cancer cells lacking in vimentin, a type of cytoskeleton proteins which is a structural component in a ... This requires the use of specific antibodies to marker molecules, with fluorescent tags recognized by the instrument.[citation ... and have some of the same cell marker proteins, chromosome abnormalities, and gene mutations found in CLL. CLL/SLL MBL consist ...
Most fluorescent DNA dyes (one of exceptions is Hoechst 33342) are not plasma membrane permeant, that is, unable to pass ... Darzynkiewicz Z, Gong JP, Juan G, Ardelt B, Traganos F (1996). "Cytometry of cyclin proteins". Cytometry. 25 (1): 1-13. doi: ... green fluorescence), or in another protocol, after removal of RNA and partial DNA denaturation, to differentially stain double- ... Before analysis, the cells are usually permeabilised and treated with a fluorescent dye that stains DNA quantitatively, such as ...
1997). "Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin". Nature. 388 (6645): 882-7. Bibcode ... Cameleon is an engineered protein based on variant of green fluorescent protein used to visualize calcium levels in living ... Sensors, Engineered proteins, Fluorescent proteins, Cell imaging, Calcium signaling). ... Protein made by the cell according to this DNA information then serves as a fluorescent indicator of calcium concentration. In ...
Though weakly fluorescent in water (green emission, 520 nm) it reacts reversibly with proteins to yield a product with a strong ... "A fluorescent natural product for ultra sensitive detection of proteins in one-dimensional and two-dimensional gel ... With respect to protein staining properties there are few differences between natural and synthetic analogs. Bell, P. J. L.; ... Choi, H. -Y.; Veal, D. A.; Karuso, P. (2005). "Epicocconone, A New Cell-Permeable Long Stokes' Shift Fluorescent Stain for Live ...
One of them, green fluorescent protein, is now widely used as a marker of molecular activity. Friday Harbor is located on the ... Eventually he purified the proteins that allow the jellyfish to fluoresce green when exposed to blue light. ...
In the past, green fluorescent protein (GFP) was used to track movement inside cells. However, GFP does not light up well and ... Thus, GFP prevented long term studies of protein movement. By using quantum dots, which are more stable, researchers can now ... This potentially allows researchers to understand when and where certain proteins are made. Self-assembled quantum dots form ... Catherine J. Murphy; Eric B. Brauns; Latha Gearheart (1996). "Quantum Dots as Inorganic DNA-Binding Proteins". MRS Proceedings ...
Osamu Shimomura (下村 脩), one of the 2008 Nobel Prize in Chemistry for green fluorescent protein (GFP). Yoshito Kishi (岸 義人), ...
Other commonly used reporters include green fluorescent protein and luciferase. Targeting sequence: Expression vectors may ... Some of these tags may also allow for increased solubility of the target protein. The target protein is fused to the protein ... Plasmids used for protein expression, called expression vectors, would include elements for translation of protein, such as a ... Protein purification tags: Some expression vectors include proteins or peptide sequences that allows for easier purification of ...
Tagged with Green Fluorescent Protein Reporter Exhibits In Vitro Biological Properties Similar to Those of the Parental FeLV-A ... GaLV Envelope Protein has biomedical significance due to its utility as a viral vector in cancer gene therapy and gene transfer ... However, because gammaretroviruses are incapable of infecting non-dividing cells, the utility of GaLV envelope protein in gene ... secretable proteins. The earliest retroviral vectors were based on the Moloney murine leukemia virus (MMLV) which when ...
Starck SR, Green HM, Alberola-Ila J, Roberts RW (2004). "A general approach to detect protein expression in vivo using ... fluorescent puromycin conjugates". Chem. Biol. 11 (7): 999-1008. doi:10.1016/j.chembiol.2004.05.011. PMID 15271358. Dando, Pam ... requires morphological changes at protein level. As puromycin inhibits protein synthesis in eukaryotic cells, researchers were ... Puromycin is an antibiotic protein synthesis inhibitor which causes premature chain termination during translation. Puromycin ...
... and can be labelled with fluorescent proteins such as green fluorescent protein or the rhodamine dye tetramethylrhodamine ... Anderson, MJ; Cohen, MW (March 1974). "Fluorescent staining of acetylcholine receptors in vertebrate skeletal muscle". The ... almost half of the protein content of the venom is composed of β-bungarotoxins. The average venom yield from specimens kept on ...
Finally, balancer chromosomes carry dominant genetic markers such as genes for green fluorescent protein or enzymes that make ... which is binding double-stranded RNA and keeping it double-stranded so that it cannot be transcribed into viral proteins. The ...
Structure of cyclized green fluorescent protein.. Hofmann, A., Iwai, H., Hess, S., Pluckthun, A., Wlodawer, A.. (2002) Acta ... Green Fluorescent Protein. A, B. 246. Aequorea victoria. Mutation(s): 20 Gene Names: GFP. ... Crystals of cyclic green fluorescent protein (cGFP) engineered by the previously reported split intein technology [Iwai et al ... Crystals of cyclic green fluorescent protein (cGFP) engineered by the previously reported split intein technology [Iwai et al ...
The biggest difference between green fluorescent protein and its red analog, DsRed, is that the chromophore of DsRed has an ... Beta sheets are green, helices red and connecting loops black. In June 2003, GFP was the protein databanks (pdb) molecule of ... in green. The crystal structure of GFP was solved in 1996. It has a unique soda can shape. Eleven beta-strands make up the beta ...
A Multi-Responsive Intrinsically Disordered Protein (IDP)-Directed Green Synthesis of Fluorescent Gold Nanoclusters. ...
Optimization using a realtime monitoring of green fluorescent protein and effect on gene reporter assay. ... Green Fluorescent Proteins, Microscopy, Fluorescence, Plasmids, PPAR-beta, Recombinant Proteins, Transfection. Abstract. Gene ... Plasmid transfection in bovine cells: Optimization using a realtime monitoring of green fluorescent protein and effect on gene ... Plasmid transfection in bovine cells: Optimization using a realtime monitoring of green fluorescent protein and effect on gene ...
... including the green fluorescent protein (GFP), contains a highly conjugated imidazolidinone ring. In many fluorescent proteins ... The chromophore of fluorescent proteins, including the green fluorescent protein (GFP), contains a highly conjugated ... A conserved interaction with the chromophore of fluorescent proteins Protein Sci. 2012 Feb;21(2):171-7. doi: 10.1002/pro.762. ... In many fluorescent proteins, the carbonyl group of the imidazolidinone ring engages in a hydrogen bond with the side chain of ...
Fluorescent proteins provide the ability to visualize, track, and quantify molecules and events in living cells with high ... Green Fluorescent Proteins. Although native green fluorescent protein produces significant fluorescence and is extremely stable ... Introduction to Fluorescent Proteins. The discovery of green fluorescent protein in the early 1960s ultimately heralded a new ... Figure 2 - Green Fluorescent Protein Fluorophore Maturation. Two predominant features of the fluorescent protein fluorophore ...
Resources Learning Center Technologies Drug Discovery Kikume Green-Red Fluorescent Protein. Kikume Green-Red Fluorescent ... Fluorescent properties. Kikume Green-Red(KikGR) spectrum data files (text files). mKikume Green excitation (4K). mKikume Green ... Protein. Bright photoconvertible fluorescent protein with high sensitivity. *Photoconvertible fluorescent protein. *Monomer/ ... The fluorescent protein Kikume Green gene was isolated from the stony coral Favia favus (Kikume-ish in Japanese). ...
title = "The conformational polymorphism of the green fluorescent protein",. abstract = "Green fluorescent protein (GFPuv) has ... Tan, H, Li, Y, Chen, L, Kudoh, T, Kasai, T & Seno, M 2012, The conformational polymorphism of the green fluorescent protein, ... N2 - Green fluorescent protein (GFPuv) has been widely used as a reporter fused to individual targeting sequences. However, its ... AB - Green fluorescent protein (GFPuv) has been widely used as a reporter fused to individual targeting sequences. However, its ...
Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled ... Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled ...
... we observe the dynamics of the proton relay reaction in the protein. Protonation of a protein carboxylate group occurs on the ... Observation of excited-state proton transfer in green fluorescent protein using ultrafast vibrational spectroscopy. ... Observation of excited-state proton transfer in green fluorescent protein using ultrafast vibrational spectroscopy. ... we observe the dynamics of the proton relay reaction in the protein. Protonation of a protein carboxylate group occurs on the ...
Crystal structure of green fluorescent protein (GFP); S65T, T203(3-OMeY); ih circular permutant (50-51). ... Unified Model for Photophysical and Electro-Optical Properties of Green Fluorescent Proteins. ...
A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum. * Welfare assessment in transgenic pigs ... Feasibility and Usability of Intraoperative Fluorescent Angiography With Indocyanine Green in Penetr… ... Diagnostic Accuracy Study of Indocyanine Green for Parathyroid Perfusion Assessment. *Indocyanine Green Tracer Using in ... Optimal Dosing of IC-Green for Visualization of Rotator Cuff Vascularity Using Advanced Imaging Moda… ...
Bodybuilding schedule, green fluorescent protein. Bodybuilding schedule, green fluorescent protein - Köp legala anabola ... The green fluorescent protein, shown here from pdb entry 1gfl , is found in a jellyfish that. Green fluorescent protein (gfp) ... Green fluorescent protein. Other articles where green fluorescent protein is discussed: martin chalfie: …discovery and ... the prmt7 protein was. Green fluorescent protein (gfp) is a 26. 9 kda protein that fluoresces bright green when exposed to blue ...
OpenLink Virtuoso version 07.20.3233 as of Jun 22 2021, on Linux (x86_64-generic_glibc25-linux-gnu), Single-Server Edition (125 GB total memory ...
Purified recombinant Green Fluorescent Protein (wild type). Catalog number: Bii-rGFP-100 Product Type. Proteins & Peptides ... Purified recombinant Green Fluorescent Protein (wild type). $385.00 Add To Cart SKU: Bii-rGFP-100 Categories : Proteins and ... Green fluorescent protein (GFP) is a 27kD protein which was originally identified in the photo organs of Aequorea victoria (A. ... GFP is a naturally fluorescent protein which emits green light at a maximum wavelength of 509 nm when excited by blue or UV ...
Enkephalinergic neurons from these mice expressed enhanced green fluorescent protein (eGFP) under the control of the ... affects the expression of enkephalin-green fluorescent protein (GFP) fluorescence in neurons in mouse spinal cord sections ... transgenic mouse in which enhanced green fluorescent protein (eGFP) is expressed in enkephalinergic neurons under the control ... Morphological and physiological features of a set of spinal substantia gelatinosa neurons defined by green fluorescent protein ...
Green Fluorescent Protein. Green fluorescent protein (GFP) is a protein in the jellyfish Aequorea Victoria that exhibits green ... The protein has 238 amino acids, three of them (Numbers 65 to 67) form a structure that emits visible green fluorescent light. ... In the jellyfish, GFP interacts with another protein, called aequorin, which emits blue light when added with calcium. ...
Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin. Nature 388, 882-887 (1997). References 51 ... Exploration of fluorescent protein voltage probes based on circularly permuted fluorescent proteins. Front. Neuroeng. 2, 14 ( ... Nakai, J., Ohkura, M. & Imoto, K. A high signal-to-noise Ca2+ probe composed of a single green fluorescent protein. Nature ... Nagai, T., Sawano, A., Park, E. S. & Miyawaki, A. Circularly permuted green fluorescent proteins engineered to sense Ca2+. Proc ...
We characterize this cdh17:eGFP line, showing green fluorescent protein (GFP) expression in the pronephric and mesonephric ... Expression of Green Fluorescent Protein in Adult Animals. In zebrafish, the cdh17::eGFP reporter line labels the pronephric as ... Stage 42 cdh17:eGFP transgenic X. laevis embryos were co-immunostained with antibodies for green fluorescent protein (GFP) and ... Stage 42 cdh17:eGFP transgenic X. laevis embryos were co-immunostained with antibodies for green fluorescent protein (GFP) and ...
In infected cells, the expression of a green fluorescent protein. Posted on November 26, 2020. by akti5463 ... In infected cells, the expression of a green fluorescent protein (GFP) reporter gene inserted into the PRRSV genome ... The addition of 4 mu M CsA after infection prevented viral RNA and protein synthesis in EAV-infected cells, and CsA treatment ...
Purification of Green Fluorescent Protein Kits 9 Products $28.90 - $122.55 Quick View ...
Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy.. Patterson GH, Knobel SM, Sharif ... Fluorescent reporter proteins that allow repeated switching between a fluorescent and a non-fluorescent state in response to ... The green fluorescent protein.. Tsien RY., Annu. Rev. Biochem. 67(), 1998 PMID: 9759496 ... The fluorescence dynamics of single molecules of green fluorescent protein. Peterman, J. Phys. Chem 103(), 1999 ...
Green Fluorescent Proteins. *. Daniel Spergel, PhD. Associate Research Scientist in Neuroscience. Research Interests ...
Red fluorescent protein (RFP)-based GECIs have inherent advantages relative to green fluorescent protein-based GECIs due to the ... Red fluorescent protein (RFP)-based GECIs have inherent advantages relative to green fluorescent protein-based GECIs due to the ... Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat ... The green fluorescent protein filter set (excitation 480/10 nm, 495 nm long pass dichroic mirror, emission 525/50 nm) and the ...
CoV, coronavirus; GFP, green fluorescent protein; Rc/Rf, chimera of Rhinolophus cornutus and R. ferrumequinum; SARS, severe ... Cells expressing each host-origin angiotensin-converting enzyme 2 were inoculated with VSV pseudotyped with spike proteins of ...
While there are many ways to label and track cells each with strengths and weakness, the green fluorescent protein (GFP) is a ... Generation of transgenic mesenchymal stem cells expressing green fluorescent protein as reporter gene using no viral vector in ... Generation of transgenic mesenchymal stem cells expressing green fluorescent protein as reporter gene using no viral vector in ... Highest transfection rate indicating green flourecscent MSC were found when the cells were transfected with 1.2 µg DNA: 2.2 µL ...
Roger Tsien was honored for developing colorful dyes called Green Fluorescent Protein. Tsien is using GFPs in the fight against ... Roger Tsien was honored for developing colorful dyes called Green Fluorescent Protein. Tsien is using GFPs i. ... Roger Tsien was honored for developing colorful dyes called Green Fluorescent Protein. The dyes come from the molecules that ...
At each step, a fluorescent image of the fluophore-labeled biotin, the avidin, and the green fluorescent protein (GFP) was ... At each step, a fluorescent image of the fluophore-labeled biotin, the avidin, and the green fluorescent protein (GFP) was ... At each step, a fluorescent image of the fluophore-labeled biotin, the avidin, and the green fluorescent protein (GFP) was ... At each step, a fluorescent image of the fluophore-labeled biotin, the avidin, and the green fluorescent protein (GFP) was ...
Production of green fluorescent protein by the methylotrophic bacterium methylobacterium extorquens ... Recombinant Proteins; Support,Non-U.S.Govt; Transformation,Bacterial. Abstract. The production of green fluorescent protein ( ... Green fluorescent protein expression; growth & development; Lac Operon; Luminescent Proteins; metabolism; methods; ... Recombinant Proteins)RN - 147336-22-9 (green fluorescent protein)RN - 7440-50-8 (Copper)RN - EC 1.13. (Oxygenases)RN - EC 1.14. ...
  • UI - 20564155LA - engRN - 0 (Culture Media)RN - 0 (Genetic Vectors)RN - 0 (Luminescent Proteins)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 147336-22-9 (green fluorescent protein)RN - 7440-50-8 (Copper)RN - EC 1.13. (
  • He is now trying to use these luminescent proteins to tag cells in humans and begin to think about ways to design cancer treatments using these proteins as an escort for targeted medicines. (
  • Plasmid transfection in bovine cells: Optimization using a realtime monitoring of green fluorescent protein and effect on gene reporter assay. (
  • Although the gene for green fluorescent protein was first cloned in 1992, the significant potential as a molecular probe was not realized until several years later when fusion products were used to track gene expression in bacteria and nematodes. (
  • The fluorescent protein Kikume Green gene was isolated from the stony coral Favia favus (Kikume-ish in Japanese). (
  • 1994) Green fluorescent protein as a marker for gene expression. (
  • Enkephalinergic neurons from these mice expressed enhanced green fluorescent protein (eGFP) under the control of the preproenkephalin (PPE) gene ( penk1 ) promoter. (
  • We observed that forskolin, an activator of adenylate cyclase (AC), affects the expression of enkephalin-green fluorescent protein (GFP) fluorescence in neurons in mouse spinal cord sections transfected with the GFP gene driven by the PPE promoter [ 16 ]. (
  • IMSEAR at SEARO: Generation of transgenic mesenchymal stem cells expressing green fluorescent protein as reporter gene using no viral vector in caprine. (
  • While there are many ways to label and track cells each with strengths and weakness, the green fluorescent protein (GFP) is a reporter gene commonly employed. (
  • Fluorescent protein tags are fundamental tools used to visualize gene products and analyze their dynamics in vivo. (
  • He searched an online database and was shocked to find a new paper in the journal Gene on the cloning of the protein - by Dr. Prasher. (
  • How do you figure out how to alter a gene so that it makes a usefully different protein? (
  • The brilliant hues are produced by a fluorescent protein gene which occurs naturally in some marine organisms. (
  • The DEP tweezer has been tested using transfected HEI 193 human schwannoma cells, with visual identification of the target cells being aided by labeling the incorporated gene product with a green fluorescent protein. (
  • In fact, in people with mutations in the gene that encodes this protein, neurons fail to migrate properly during development. (
  • Among the signaling pathways regulated by GSK3s, the Wnt canonical pathway is the most well described, with GSK3β inhibition triggering an increase in β -catenin protein levels and its nuclear translocation to activate target gene expression ( Doble and Woodgett, 2003 ). (
  • E), nucleocapsid protein (N), RNA-dependent RNA drome coronavirus-2 (SARS-CoV-2) causes coronavirus polymerase enzyme, and ORF1 gene) (4-6) either by disease 2019 (COVID-19), which was declared a pandemic nucleic acid amplification testing or detection of virus- on 11 March 2020, because of its rapid spread around the specific proteins by antigen testing (7,8) . (
  • The capsids comprise multiple copies of one or more identical CP subunits, thus providing many different possibilities for the selective attachment and presentation of numerous organic and inorganic molecules, including metals, semi-conductors, carbohydrates, peptides and larger proteins such as antibodies. (
  • Roger Tsien was honored for developing colorful dyes called Green Fluorescent Protein. (
  • They used genetic engineering to exchange individual moieties of building blocks and modified the protein chemically using fluorescent dyes. (
  • abstract = "Preferential immobilization of a protein by using a nano-patterned organosilane monolayer as the template was investigated. (
  • During recovery is when most muscle is built, because muscle protein synthesis increases by 50% four hours after a workout (like resistance training). (
  • The addition of 4 mu M CsA after infection prevented viral RNA and protein synthesis in EAV-infected cells, and CsA treatment check details resulted in a 2.5- to 4-log-unit reduction of PRRSV or EAV infectious progeny. (
  • Several phosphorylation sites in the N-terminal regions of Orm proteins played crucial roles in the course of sphingolipid synthesis. (
  • ORMDL3 V1 open reading frame was subcloned into pEGFP-C1 vector and it was found that the protein synthesis had been followed in transfected Hela cells. (
  • Hormone replacement therapy that induces the synthesis of DNA, RNA, and various proteins in target tissues. (
  • RNA, including ribosomes, tRNA and mRNA, are in shades of magenta, and protein synthesis factors are in purple. (
  • Enzymes are in shades of blue, with a cooler cobalt blue for enzymes interacting with the protein synthesis machinery, and metabolic enzymes in shades of turquoise. (
  • In addition to standard oligos synthesis, we also provide scientific research services such as long oligos synthesis, phosphorothioate oligos (S-Oligo) synthesis, modified oligos synthesis, fluorescent oligos synthesis, and real-time quantitative PCR probes to meet your different needs. (
  • green fluorescent protein (gfp) is a β-barrel-shaped protein consisting of 238 amino acids and with a molecular weight of ~27kda. (
  • The predicted protein sequences of this isoform lacked 59 amino acids in the N-terminus of the wild-type ORMDL3 protein. (
  • The discovery of green fluorescent protein in the early 1960s ultimately heralded a new era in cell biology by enabling investigators to apply molecular cloning methods, fusing the fluorophore moiety to a wide variety of protein and enzyme targets, in order to monitor cellular processes in living systems using optical microscopy and related methodology. (
  • When coupled to recent technical advances in widefield fluorescence and confocal microscopy, including ultrafast low light level digital cameras and multitracking laser control systems, the green fluorescent protein and its color-shifted genetic derivatives have demonstrated invaluable service in many thousands of live-cell imaging experiments. (
  • Mizuno H, Dedecker P, Ando R, Fukano T, Hofkens J, Miyawaki A Higher resolution in localization microscopy by slower switching of a photochromic protein. (
  • Fluorescent reporter proteins that allow repeated switching between a fluorescent and a non-fluorescent state in response to specific wavelengths of light are novel tools for monitoring of protein trafficking and super-resolution fluorescence microscopy in living organisms. (
  • In separate experiments this study initially compared white versus fluorescent light microscopy for counting A. fumigatus conidia, then compared fluorescent microscopy counting of corresponding filter halves, and finally compared qPCR enumeration to counting by fluorescent light microscopy. (
  • Filters (n = 38) loaded with GFP conidia in the aerosol chamber were divided in half and analyzed by either fluorescent microscopy or qPCR. (
  • The presence of iLOV fusion proteins and hybrid particles was confirmed by western blot analysis and transmission electron microscopy. (
  • In June 2003, GFP was the protein databank's (pdb) molecule of the month . (
  • Gfp stands for green fluorescent protein (the official name for the molecule) and is, imaginatively, a protein that fluoresces green in the presence of uv. (
  • In the laboratory, the GFP protein has been used extensively as a reporter molecule to label, and study, cellular and subcellular proteins in living cells using a wide range of biological applications, including oncology, cardiovascular diseases, brain research, embryology and plant sciences, just to name a few. (
  • For example, expression of a constitutively active form of a tagged protein or knockdown of a target by RNAi can simulate the effects of a sought-after small molecule in a screen. (
  • Intraembryonal injection of adenoviral vectors into progenitor cells of the forebrain yielded an even more widespread distribution of a fluorescent green protein used as a tracking molecule. (
  • By fusing the jellyfish enhanced green fluorescent protein reporter molecule (EGFP) to the carboxy-terminus of Us9, we demonstrated that Us9 not only is capable of targeting a Us9-EGFP fusion protein to the Golgi compartment, it also is able to direct efficient incorporation of such chimeric molecules into infectious viral particles. (
  • Since 2000, the RCSB PDB Molecule of the Month series has introduced millions of visitors to the shape and function of the 3D structures archived in the Protein Data Bank. (
  • The fluorescent protein technique avoids the problem of purifying, tagging, and introducing labeled proteins into cells or the task of producing specific antibodies for surface or internal antigens. (
  • For the quantitation of the expression of a specific protein, tagged with GFP in these model systems, antibodies to GFP have proven to be of value in immunoblotting studies and ELISA protocols. (
  • Americans Martin Chalfie and Roger Tsien and Japan s Osamu Shimomura discovered and successfully developed a fluorescent protein found in jellyfish. (
  • CoralHue® Kikume Green-Red1 (KikGR1) protein emits bright green fluorescence that can be irreversibly converted to red. (
  • Green fluorescent protein (gfp), a 27 kda protein derived from the jellyfish aequorea victoria, emits green light (emission peak 509 nm) when excited by. (
  • GFP is a naturally fluorescent protein which emits green light at a maximum wavelength of 509 nm when excited by blue or UV light. (
  • It emits green fluorescence when bound to DNA or RNA. (
  • Mutants of asfp595 (also called the "kindling fluorescent protein" or kfp) are reversibly photoswitchable, in that exposure to intense green light can. (
  • Here, we describe variants of the reversibly photoswitchable fluorescent proteins rsFastLime, bsDronpa, and Padron that have been codon-optimized for the use in transgenic Arabidopsis plants. (
  • Because the new reversibly photoswitchable fluorescent proteins show an increase in signal strength during each photoactivation cycle, we were able to generate a large number of scans of the same region and reconstruct 3-D images of AtGRP7 expression in the root tip. (
  • Lummer M, Humpert F, Wiedenlübbert M, Sauer M, Schüttpelz M, Staiger D. A new set of reversibly photoswitchable fluorescent proteins for use in transgenic plants. (
  • Western blot analysis detected a ∼38 kDa EGFP-ORMDL3 V1 fusion protein. (
  • Through internalization assays with an EGFP epitope-tagged Us9 protein, we demonstrate that the maintenance of Us9 to the TGN region is a dynamic process involving retrieval of molecules from the cell surface. (
  • Osamu Shimomura and Frank Johnson, working at the Friday Harbor Laboratories of the University of Washington in 1961, first isolated a calcium-dependent bioluminescent protein from the Aequorea victoria jellyfish, which they named aequorin . (
  • During the isolation procedure, a second protein was observed that lacked the blue-emitting bioluminescent properties of aequorin, but was able to produce green fluorescence when illuminated with ultraviolet light. (
  • Over the next two decades, researchers determined that aequorin and the green fluorescent protein work together in the light organs of the jellyfish to convert calcium-induced luminescent signals into the green fluorescence characteristic of the species. (
  • In A.victoria, calcium ions bind and activate the protein aequorin causing the release of blue fluorescence, which is then absorbed by GFP resulting in the release of green fluorescence. (
  • He found that calcium ions activate a jellyfish protein called aequorin. (
  • Once activated, aequorin produces blue light, but if the green fluorescent protein is nearby, the two proteins together yield a bright green signal. (
  • At that time, they called this protein aequorin. (
  • This distinct discovery happened in a very interesting chance and then, because he was studying the how to make this protein Aequorin to have light, at that time, and then he tried so many solutions and the combinations and he failed. (
  • But one day, when he was about to finish his experiment, he throw out his solutions into the sink, and then, of course, his left over Aequorin, this protein into sink as well, so all of a sudden, he saw in the sink the protein has very strong light over there. (
  • And then the first time, he realized this protein, Aequorin here, might be very useful for biological studies and research. (
  • At each step, a fluorescent image of the fluophore-labeled biotin, the avidin, and the green fluorescent protein (GFP) was obtained by fluoroscent microscope. (
  • A reliable taxonomic character, distributional pattern of green fluorescent protein (GFP), that is contributable to species demarcation by observing living materials under the fluorescent microscope, has not been much done in hydroids in contrast to hydromedusae. (
  • Produced in 2011, by the National Institute of Allergy and Infectious Diseases (NIAID), this photomicrographic image, created using a fluorescent light equipped microscope, depicts what were red-colored mouse macrophages, 12-hours after having been inoculated with a virulent strain of green glowing, Francisella tularensis bacteria. (
  • Chlorophytas are recognised as green structures with compartments grouped into chains when seen under a microscope. (
  • They feature a large, elongated green structure under the microscope. (
  • By combining the 'positively switchable' PADRON C-s with the 'negatively switchable' rsFastLIME-s or bsDRONPA-s, two different fluorescent reporter proteins could be imaged at the same wavelength upon transient expression in Nicotiana benthamiana cells. (
  • Most quantitative comparisons of the brightness and photostability of different fluorescent proteins have been made in vitro, removed from biological variables that govern their performance in cells or organisms. (
  • We generated transgenic Caenorhabditis elegans strains expressing green, yellow, or red fluorescent proteins in embryos and imaged embryos expressing different fluorescent proteins under the same conditions for direct comparison. (
  • Applying an electroporation technique, in vitro synthesized Green Fluorescent Protein (GFP) mRNA was transfected as an indicator into the DCs derived from a healthy donor. (
  • We found that mNeonGreen was not as bright in vivo as predicted based on in vitro data but is a better tag than GFP for specific kinds of experiments, and we report on optimal red fluorescent proteins. (
  • However, the experiments in the study used metabolically labeled proteins in the supernatant of in vitro cultured P. knowlesi , and the binding activity was qualitatively determined by probing the 135 kD protein with in situ erythrocytes and electrophorectically-separated erythrocyte proteins on blots. (
  • Follow-up experiments demonstrated that EJ exhibited a good inhibitory effect on cancer cell metastasis both in vitro and in vivo, and could effectively reduce the expression of STAT3, MMP-2, and MMP-9 proteins in cells, while the knockdown of STAT3 could weaken the inhibitory effect of EJ on cancer cell metastasis. (
  • Strong in vitro cell-mediated responses to various retinal autoantigens, including self-antigen (S-Ag) and interphotoreceptor retinoid-binding protein (IRBP), have been observed in patients with birdshot chorioretinopathy. (
  • This allows the formation of recombinant proteins essential for the development of relevant in-vitro and in-vivo methodologi. (
  • It's been 25 years since researchers coaxed a bacterium to synthesize an unusual jellyfish protein that fluoresced bright green when irradiated with blue light. (
  • This is supported by the absence of crystal contacts in the linker-peptide region and the fact that the core of the protein exhibits a very similar conformation to that known from other GFP structures, thereby not implicating any constraints arising from the presence of the artificial linker. (
  • A survey of high-resolution structures of fluorescent proteins indicates that electron lone pairs of a main-chain oxygen-Thr62 in GFP-donate electron density into an antibonding orbital of the imidazolidinone carbonyl group. (
  • mKikGR1 can be used for labeling proteins or subcellular structures. (
  • In addition to distinct organ-specific expression pattern, three CYP76AHs exhibited different genomic structures (w or w/o introns), low protein sequence identities (51-63%) and were placed in separate subclades in the phylogenetic tree. (
  • Background fluorescence in the red channel illustrates basic anatomy and structures of the brain, and the injection site and projections are shown in the green channel. (
  • The cytoskeleton is made from protein structures called microtubules, made visible by fluorescently tagging a protein called doublecortin (orange). (
  • Within months, another group had also fused this small green fluorescent protein (GFP) to larger proteins to make their whereabouts inside the cell come to light-like never before. (
  • Is the Subject Area "Peripheral membrane proteins" applicable to this article? (
  • The HeLa cells were co-transfected with sub-cellular localization vectors fused to cyan ( mTurquoise ) and yellow ( mVenus ) fluorescent protein coding sequences (Golgi complex and the nucleus, respectively), as well as the "Fruit" protein, mCherry, targeting the mitochondrial network. (
  • Green fluorescent protein, and its mutated allelic forms, blue, cyan, and yellow fluorescent proteins are used to construct fluorescent chimeric proteins that can be expressed in living cells, tissues, and entire organisms, after transfection with the engineered vectors. (
  • The production of green fluorescent protein (GFP) in Methylobacterium extorquens was studied by creating four different constructs using pJB3KmD, pRK310 and pVK101 vectors, as well as pLac and soluble methane monooxygenase (sMMO) promoters. (
  • Our data suggest that TMV-based vectors are suitable for the production of proteins at least as large as iLOV when combined with the FMDV 2A sequence. (
  • Studies have shown the expression of recombinant GABA(A) R subunits tagged with the green fluorescent protein (GFP), a 26.9 kDa protein that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. (
  • With the rapid evolution of fluorescent protein technology, the utility of this genetically encoded fluorophore for a wide spectrum of applications beyond the simple tracking of tagged biomolecules in living cells is now becoming fully appreciated. (
  • Illustrated in Figure 1 are two examples of multiple fluorescent protein labeling in living cells using fusion products targeted at sub-cellular (organelle) locations. (
  • Furthermore, the proteins also show strong fluorescence in mammalian cells including COS-7 and HeLa cells. (
  • Cells expressing each host-origin angiotensin-converting enzyme 2 were inoculated with VSV pseudotyped with spike proteins of Rc-o319 (A), SARS-CoV (B), SARS-CoV-2 (C), or glycoprotein of VSV (D). At 20 hour postinfection, GFP-positive cells were counted and the infectivity titers were calculated. (
  • Highest transfection rate indicating green flourecscent MSC were found when the cells were transfected with 1.2 µg DNA: 2.2 µL lipofectamine and 1.5 µg DNA: 2.5 µL lipofectamine than other combinations. (
  • Since they were discovered to be released from sheep reticulocytes, exosomes were once defined as unwanted proteins secreted from the cells and manifested as a membrane vesicle [ 8 ]. (
  • He is one of three scientists to win the 2008 Nobel Prize in Chemistry for their work in developing green fluorescent protein (GFP) to track changes in cells or organisms. (
  • Infectivity of retroviral severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike protein (S) pseudotypes on target cells. (
  • Murine leukemia virus (MLV) or HIV pseudotypes bearing either the pantropic vesicular stomatitis virus envelope protein (VSV-G) as a positive control, or the SARS-CoV S, were added to target cells. (
  • After 72 h, green fluorescent protein (GFP)-positive cells were counted by fluorescence-activated cell sorter analysis. (
  • Proteins can be effectively expressed by transduced retroviruses, but this strategy is limited by the ability of retroviruses to infect only dividing cells. (
  • Adenoviruses are believed to have a lower transforming potential, but they can mediate expression of exogenous proteins only in an acute, transient fashion, as they do not integrate into the genome of the host cells. (
  • To define mechanisms involved, we treated established desmoplastic pancreatic tumors with CAR T cells directed to fibroblast activation protein (FAP), an enzyme highly overexpressed on a subset of cancer-associated fibroblasts (CAFs). (
  • After cell extension, the stably pMIG-infected cells had been isolated using stream cytometry analysis regarding to green fluorescent proteins expression. (
  • The untreated cells fail to control replication of F. tularensis , as evidenced by their contents of the green bacteria. (
  • The cutaneous myeloma cells stained in a characteristic fashion with methyl green-pyronin and touch imprints of the tumors stained with Wright's stain demonstrated, however, typical myeloma cells. (
  • In cryostat-sectioned specimens from this tumor, these fluorescent cells appeared to buttress the tumor masses in the skin. (
  • We provide furthermore evidence that endogenous ezrin and moesin colocalize with F-actin at the contracted tail of polarized cells, similar to ectopically expressed green fluorescent protein-tagged ezrin. (
  • The molecular mechanisms that enable the virus to invade and spread in the nervous systems of so many different animals are not known, but it is well known that the virion envelope proteins gE and gI are required in almost every case studied for efficient cell-to-cell spread both in non-neuronal and neuronal cells. (
  • In healthy eyes, the cells that make up blood vessels are bound together by proteins residing on the surface of the cell that are directed into place by Tie 2, another protein. (
  • Tie2 proteins pack tightly together where cells meet their neighbors and act like Velcro to create a fluid-tight connection between cells in the blood vessel's wall. (
  • When they added the AXT107 drug to these cells, the researchers found that AXT107 initiated a series of changes to cellular proteins. (
  • Groups of Tie2 proteins began to congregate where cells met their neighbors, and began rebuilding connections with other blood vessel cells. (
  • So they grew the cells in a single layer and tested whether fluid could pass through by pouring a fluorescent liquid on top of the cells and checking to see if any of the glowing liquid ended up underneath. (
  • Host-guest interactions of curcubiturils with fluorescent probes, photoCORMs, TTA-UC molecular systems, Au(I) - coumarin complexes, supramolecular chemosensors, and ligands aiming to to inhibit DNA telomerase in cancer cells are also being explored. (
  • And so, the researchers tagged doublecortin with an orange fluorescent protein, engineered its expression in the breast cancer cells, and van Haren started taking pictures. (
  • This video is certainly a good example of the illuminating power of fluorescent proteins: enabling us to see cells and their cytoskeletons as incredibly dynamic, constantly moving entities. (
  • (C-D) Confocal images of TNBC cell lines treated with panobinostat (100 nM) or vehicle for 18 hours, fixed, permeabilized and stained red (rhodamine phalloidin) for actin filaments and green (Alexa Fluor® 488) for acetyl-histones (C) H3 (Lys9) or (D) H4 (Lys8). (
  • Filaments of another protein called actin (purple) are seen here as the fine meshwork in the cell periphery. (
  • Other proteins present in exosomes are the ones associated with lipid rafts, including glycosyl phosphatidylinositol-anchored proteins, heat shock, and proteins related to multi-vesicular body (MVB) biogenesis (eg, TSG101 and Alix) (2). (
  • Use of green fluorescent protein-expressing Aspergillus fumigatus conidia to validate quantitative PCR analysis of air samples collected on filters. (
  • This study used green fluorescent protein (GFP)-expressing Aspergillus fumigatus conidia to compare quantitative PCR (qPCR) enumeration with direct epifluorescent microscopic filter counts of conidia collected on filters in a test chamber. (
  • CoralHue® monomeric Kikume Green-Red (mKikGR1) maintains the brightness of the parent protein KikGR1. (
  • Protein engineering efforts have yielded three major lineages of monomeric red fluorescent proteins (RFPs) derived from their naturally oligomeric precursors (Fig. 1a ). (
  • Together, these three lineages of monomeric RFPs are commonly used in a variety of fluorescence imaging applications and have served as templates for developing red fluorescent indicators of various biochemical activities [ 13 ]. (
  • The monomeric non-oxygen-dependent fluorescent protein iLOV can be used as an alternative to green fluorescent protein. (
  • 2) GFPuv could reduce its partner's polymorphism as a fusion protein. (
  • Upon photoactivation of the AtGRP7:rsFastLIME-s fusion protein in a defined region of a transgenic Arabidopsis root, spreading of the fluorescence signal into adjacent regions was observed, indicating that movement from cell to cell can be monitored. (
  • but to our knowledge it has not yet been used in a TMV CP fusion protein. (
  • The biggest difference between green fluorescent protein and its red analog, DsRed, is that the chromophore of DsRed has an extra double bond (drawn in yellow) which extends the chromophores conjugation and causes the red-shift. (
  • The chromophore of fluorescent proteins, including the green fluorescent protein (GFP), contains a highly conjugated imidazolidinone ring. (
  • Accordingly, this n→π* interaction merits inclusion in computational and photophysical analyses of the chromophore, and in speculations about the molecular evolution of fluorescent proteins. (
  • KikGR1 contains a His62-Tyr63-Gly64 tripeptide sequence which forms a green chromophore that can be photoconverted to a red one via formal beta-elimination and subsequent extension of the π-conjugated system. (
  • In addition to the expected bleaching and transient infrared absorption of bands associated with the chromophore, we observe the dynamics of the proton relay reaction in the protein. (
  • In this study, the iLOV sequence was genetically fused either directly or via a glycine-serine linker to the C-terminus of the Tobacco mosaic virus (TMV) coat protein (CP) and also carried an N-terminal Foot-and-mouth disease virus (FMDV) 2A sequence. (
  • Using the protein sequences of these receptors as queries we identified overlapping expressed sequence tags which were predicted to encode an additional subtype. (
  • In mammalian cell based models of both polyglutamine and polyalanine diseases, the mutant proteins are much more prone to aggregate formation than their wild-type counterparts and cause significantly more cell death. (
  • Our previous studies suggested that mammalian heat shock proteins might be able to play similar roles in both diseases. (
  • These advances expand the potential of in vivo imaging experiments and facilitate experimentation with new, bright, photostable fluorescent proteins. (
  • These results identify ideal fluorescent proteins for imaging in vivo in C. elegans embryos and suggest good candidate fluorescent proteins to test in other animal model systems for in vivo imaging experiments. (
  • The other two are well-known lab workhorses: the LacI protein from the E. coli bacterium and the green fluorescent protein (GFP) used as a marker in biology experiments. (
  • To understand how the drug they developed could strengthen these connections, the researchers designed a series of experiments to explore how AXT107 affects the control of Tie2 and the Velcro-like proteins. (
  • The Green Fluorescent Protein (GFP), from the jellyfish Aequorea Victoria, is a vital imaging tool in cellular and molecular biology. (
  • Extracellular vesicles have a bilayer membrane structure and serve as vehicles to deliver different kinds of cellular cargo, including proteins, lipids, nucleic acids, and receptors (1). (
  • The steady-state residence of the Us9 protein is in a cellular compartment in or near the trans -Golgi network (TGN). (
  • Mimicking enzyme function and increasing performance of naturally evolved proteins is one of the most challenging and intriguing aims of nanoscience. (
  • Cytoskeleton offers several reagents for live-cell research including fluorescent proteins, cell permeable protein activators and inhibitors, as well as our recent addition of live cell imaging probes. (
  • Comparative assessment of fluorescent proteins for in vivo imaging in an animal model system. (
  • To address the gap, we quantitatively assessed fluorescent protein properties in vivo in an animal model system. (
  • Zusätzlich wurden die Protein-Protein Interaktionen der akzessorischen Gasvesikelproteine und GvpA direkt in vivo mittels split-GFP untersucht. (
  • Though the researchers are using a breast cancer cell line, their primary interest is in the doublecortin protein, which is normally found in association with microtubules in the developing brain. (
  • Since these early studies, green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors that are broadly referred to as fluorescent proteins. (
  • So in the luciferin system, that we need to have both protein that we called the luciferin, and enzyme, that we called luciferase and oxygen. (
  • Red fluorescent proteins have been isolated from other species, including coral reef organisms, and are similarly useful. (
  • Recent advances in genome editing have expedited the precise insertion of fluorescent protein tags into the genomes of diverse organisms. (
  • Some organisms deposit green pigments inside their cell walls because they are photosynthetic. (
  • Glycogen synthase kinase 3 (GSK3) proteins (GSK3α and GSK3β) are key mediators of signaling pathways, with crucial roles in coordinating fundamental biological processes during neural development. (
  • Green fluorescent protein (GFP) is a 27kD protein which was originally identified in the photo organs of Aequorea victoria (A. victoria) jellyfish. (
  • Protein analogs and derivatives of the Aequorea victoria green fluorescent protein that emit light ( FLUORESCENCE ) when excited with ULTRAVIOLET RAYS . (
  • He found lots of calcium or calcium solutions there, and then he found with the help of calcium ions that this protein can have light, fluorescence, so in the sea water, in jellyfish, because in our sea water, the calcium concentration is pretty high. (
  • What if a bioluminescent protein could cast its light in an organism that was already transparent? (
  • Green fluorescent protein (GFPuv) has been widely used as a reporter fused to individual targeting sequences. (
  • I work with proteins, so I've done Western blots throughout my career. (
  • Fluorescent protein fusions have been instrumental for the tagging of plant virus particles. (
  • Furthermore, we discovered that children and adults who got KD during years as a child without developing CAA didn't react to the Fc proteins antigens and play a central part in keeping immunological tolerance [1,2]. (
  • DI-fusion Light-triggered green fluorescent protein silencing in human. (
  • 9 kda protein that fluoresces bright green when exposed to blue or ultraviolet light. (
  • Red fluorescent protein (RFP)-based GECIs have inherent advantages relative to green fluorescent protein-based GECIs due to the longer wavelength light used for excitation. (
  • The same green light occurs if GFP alone is illuminated with ultraviolet or blue light. (
  • Finally, the interaction of light with soluble proteins disclosed that the domain assembles in two discrete steps. (
  • We only need this protein, change the information, and then we can have the light, we can have the visible light. (
  • Jellyfish will glow under blue and ultraviolet light because of this protein which the three scientists have become known for. (
  • In addition, compounds that directly affect either Ca channels or proteins that modulate their activity are used to treat a number of cardiovascular and neurological pathologies ( Miller, 2001 ). (