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...
For example, a gene from a jellyfish, encoding a fluorescent protein called GFP, or green fluorescent protein, can be ... "Green fluorescent protein takes Nobel prize". Lewis Brindley. Retrieved 2015-05-31.. ... Scientists have genetically engineered several organisms, including some mammals, to include green fluorescent protein (GFP), ... "Green fluorescent protein (GFP) as a marker of aryl hydrocarbon receptor (AhR) function in developing zebrafish (Danio rerio)" ...
In 1961, Osamu Shimomura extracted green fluorescent protein (GFP) and another bioluminescent protein, called aequorin, from ... where the green fluorescent protein, used by some species to cause bioluminescence, has been adapted as a fluorescent marker ... Further information: Bioluminescence and Green fluorescent protein. Pliny the Elder reported in his Natural History that the ... The hydromedusa Aequorea victoria was the source of green fluorescent protein, studied for its role in bioluminescence and ...
"Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins". Nature. 394 (6689): 192-195. doi ... postdoctoral research with Harvey Lodish at Massachusetts Institute of Technology working on glycosylation of membrane proteins ...
"The Green Fluorescent Protein". Annual Review of Biochemistry (Annual Reviews) 67 (1): 509-544. doi:10.1146/annurev.biochem. ... He was awarded the 2008 Nobel Prize in Chemistry for his discovery and development of the green fluorescent protein with ...
... "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, Martin (1995-10-01). "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. Chalfie's lab Website Martin Chalfie on Nobelprize. ... 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 ...
... 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". "The Nobel Prize in Chemistry 2008". Retrieved 2020-10-19. v ... 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 ... 2011). "Bright and stable near infra-red fluorescent protein for in vivo imaging". Nat Biotechnol. 29 (8): 757-761. doi:10.1038 ... Jellyfish- and coral-derived fluorescent proteins require oxygen and produce a stoichiometric amount of hydrogen peroxide upon ...
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 ...
... and is coupled with a closely interacting green fluorescent protein (RrGFP), and a Ca++ activated luciferin binding protein ( ... Loening, A. M.; Fenn, T. D.; Gambhir, S. S. (2007). "Crystal Structures of the Luciferase and Green Fluorescent Protein from ... Characterization of the Renilla green-fluorescent protein". The Journal of Biological Chemistry. 254 (3): 781-88. PMID 33175. ... Ward, William W.; Cormier, Milton J. (April 1978). "Energy Transfer Via Protein-Protein Interaction in Renilla Bioluminescence ...
Green fluorescent protein (GFP). NotesEdit. *. Penner-Hahn, James E. (2013). "Chapter 2. Technologies for Detecting Metals in ... The cell membrane consists of lipids and proteins which accounts for its hydrophobicity as a result of being non-polar ... The main constituents of the general molecular composition of the cell includes: proteins and lipids which are either free ...
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.. ... As puromycin inhibits protein synthesis in eukaryotic cells, researchers were able to show that injections of this drug will ... Long-term synaptic plasticity, such as is required for memory processes, requires morphological changes at protein level. ...
Transgenic products in vivo, particularly the green fluorescent protein or related fluorescent proteins ... Concentrations of a protein of interest in the samples can be obtained by comparing the fluorescent signals to those of a ... Protein modifications, phospho-proteins. *Scattering of light can be used to measure volume (by forward scatter) and ... green-HeNe, HeCd (UV)); diode lasers (blue, green, red, violet) resulting in light signals. The detector and analog-to-digital ...
It also exhibits proteins similar to green fluorescent protein. Norzoanthamine (2018). Zoanthus Lamarck, 1801. Accessed through ...
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 Deoxyribozyme Anti-thrombin aptamers ... 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 ...
"Green fluorescent protein takes Nobel prize". Lewis Brindley. Retrieved 2015-05-31. 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 ...
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.). 515. pp. 165-175. doi:10.1007/ ... 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 ...
"Application of fluorescence lifetime imaging of enhanced green fluorescent protein to intracellular pH measurements". ... family proteins. FLIM has been used in clinical multiphoton tomography to detect intradermal cancer cells as well as ... has also been used to show the interaction of both types of nuclear intermediate filament proteins lamins A and B1 in distinct ... "Fluorescence Lifetime Imaging of Free and Protein-Bound NADH". Proceedings of the National Academy of Sciences of the United ...
... 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 ...
... yellow fluorescent protein (YFP) pair. Both are color variants of green fluorescent protein (GFP). Labeling with organic ... Bevan N, Rees S (2006). "Pharmaceutical Applications of GFP and RCFP". In Chalfie M, Kain SR (eds.). Green Fluorescent Protein ... such as protein-protein interactions, protein-DNA interactions, and protein conformational changes. For monitoring the complex ... FRET can be used to observe membrane fluidity, movement and dispersal of membrane proteins, membrane lipid-protein and protein- ...
"for the discovery and development of the green fluorescent protein, GFP". *^ "The Nobel Prize in Chemistry 2009". Nobelprize. ... "for his discovery that enzymes can be crystallized; for their preparation of enzymes and virus proteins in a pure form"". ... "for his work on the structure of proteins, especially that of insulin". ... especially for his discoveries concerning the complex nature of the serum proteins". ...
In the animals, 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): 882-87. doi:10.1038/ ... Shimomura O (2005). "The discovery of aequorin and green fluorescent protein". J Microsc. 217 (Pt 1): 1-15. doi:10.1111/j.0022- ...
"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 ...
"Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin". Nature. 388 (6645): 882-7. Bibcode: ... Nakai J, Ohkura M, Imoto K (February 2001). "A high signal-to-noise Ca(2+) probe composed of a single green fluorescent protein ... where scientists have fused fluorescent proteins to detector proteins. An example of this is voltage-sensitive fluorescent ... Zhou XX, Chung HK, Lam AJ, Lin MZ (November 2012). "Optical control of protein activity by fluorescent protein domains". ...
"A highly metastatic Lewis lung carcinoma orthotopic green fluorescent protein model". Clinical & Experimental Metastasis. 18 (1 ...
It consists of a pH-sensitive form of green fluorescent protein (GFP) fused to the luminal side of a vesicle-associated ... "Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins". Nature. 394 (6689): 192-195. doi ... Synapto-pHluorin sometimes consists of yellow fluorescent protein (YFP) to monitor the cytoplasm because its pKa is higher than ... Ashby, Michael C.; Ibaraki, Kyoko; Henley, Jeremy M. (May 2004). "It's green outside: tracking cell surface proteins with pH- ...
Other green dyes include Oregon Green, Tokyo Green, SNAFL, and carboxynaphthofluorescein. These dyes, along with newer ... In plant science, fluorescein, and other fluorescent dyes, have been used to monitor and study plant vasculature, particularly ... FITC reacts with the amine groups of many biologically relevant compounds including intracellular proteins to form a thiourea ... The color of its aqueous solution varies from green to orange as a function of the way it is observed: by reflection or by ...
... commonly the green fluorescent protein or GFP) and an allele of a gene to be studied (both on chromosomes bearing FRT sites). ... The resulting BLM protein is defective. the defect in RecQ an helicase facilitates the defective unwinding of DNA during ...
The α subunits are represented as green spheres and the β subunits as protein backbones colored by individual polypeptide chain ... Fluorescent inhibitors have also been developed to specifically label the active sites of the assembled proteasome.[115] ... The protein degradation processEdit. Ribbon diagram of ubiquitin, the highly conserved protein that serves as a molecular tag ... Proteasomes are protein complexes which degrade unneeded or damaged proteins by proteolysis, a chemical reaction that breaks ...
C. Zhi, Y. Bando, C. Tang and D. Golberg : «Immobilization of Proteins on Boron Nitride Nanotubes» J. Am. Chem. Soc. 127[49] ( ... Green" Chemical Functionalization of Boron Nitride Nanotubes» JOURNAL OF PHYSICAL CHEMISTRY C 111[50] (2007) 18545-18549 ... Boron nitride nanotubes as vehicles for intracellular delivery of fluorescent drugs and probes» Nanomedicine 11[5] (2016) 447- ... Novel polymer nanocomposites from bioinspired green aqueous functionalization of BNNTs» Polym.Chem 3[4] (2012) 962-969 DOI: ...
... and can be labelled with fluorescent proteins such as green fluorescent protein or the rhodamine dye tetramethylrhodamine ... based on the amino acid sequences of venom proteins". Molecular Phylogenetics and Evolution. 8 (3): 349-62. CiteSeerX 10.1. ... almost half of the protein content of the venom is composed of β-bungarotoxins.[19] ... "Fluorescent staining of acetylcholine receptors in vertebrate skeletal muscle". The Journal of Physiology. 237 (2): 385-400. ...
"for the discovery and development of the green fluorescent protein, GFP"[101] ... "for their preparation of enzymes and virus proteins in a pure form"[38] ... "for his work on the structure of proteins, especially that of insulin"[50] ... especially for his discoveries concerning the complex nature of the serum proteins"[40] ...
In whole milk, 14% of the flavins are bound noncovalently to specific proteins.[15] Egg white and egg yolk contain specialized ... Because riboflavin is fluorescent under UV light, dilute solutions (0.015-0.025% w/w) are often used to detect leaks or to ... Further, the researchers noted that a yellow-green fluorescence in each extract promoted rat growth, and that the intensity of ... Free riboflavin is naturally present in foods along with protein-bound FMN and FAD. Bovine milk contains mainly free riboflavin ...
Lareau LF, Green RE, Bhatnagar RS, Brenner SE (June 2004). "The evolving roles of alternative splicing". Current Opinion in ... McClain WH (November 1993). "Rules that govern tRNA identity in protein synthesis". Journal of Molecular Biology. 234 (2): 257- ... fluorescent, biotinylated, and redox-active amino acids.[14] Another use is introducing amino acids bearing reactive functional ... For instance, one can start with the gene for a protein that binds a certain sequence of DNA, and, by directing an unnatural ...
Proteins in different cellular compartments and structures tagged with green fluorescent protein (here, white) ... Main article: Protein domain. Many proteins are composed of several protein domains, i.e. segments of a protein that fold into ... such as green fluorescent protein (GFP).[46] The fused protein's position within the cell can be cleanly and efficiently ... globular proteins, fibrous proteins, and membrane proteins. Almost all globular proteins are soluble and many are enzymes. ...
positive regulation of protein secretion. · regulation of axonogenesis. · regulation of synaptic transmission. · protein ... 1f8u: CRYSTAL STRUCTURE OF MUTANT E202Q OF HUMAN ACETYLCHOLINESTERASE COMPLEXED WITH GREEN MAMBA VENOM PEPTIDE FASCICULIN-II ... Mapping the human acetylcholinesterase gene to chromosome 7q22 by fluorescent in situ hybridization coupled with selective PCR ... protein self-association. 細胞成分. · extracellular region. · extracellular region. · extracellular region. · basal lamina. · ...
"Protein & Cell. 6: 363-72. doi:10.1007/s13238-015-0153-5. PMC 4417674 . PMID 25894090. Retrieved 24 April 2015.. ... Collins FS, Green ED, Guttmacher AE, Guyer MS, US National Human Genome Research Institute (2003). "A vision for the future of ... Fluorescence in situ hybridization (FISH) involves fluorescent labeling of probes that bind to specific DNA sequences, used for ... In general, only the parts of the gene that code for the expressed protein (exons) and small amounts of the flanking ...
... green fluorescent protein with the protein of interest).. Techniques used for horizontal scanning[edit]. Four types of confocal ... Green signal from anti-tubulin antibody conjugated with Alexa Fluor 488) and nuclei (blue signal from DNA stained with DAPI) in ... GFP fusion protein being expressed in Nicotiana benthamiana. The fluorescence is visible by confocal microscopy. ... Also, transgenic techniques can create organisms that produce their own fluorescent chimeric molecules (such as a fusion of GFP ...
Green fluorescent protein-Tag (GFP-Tag). *Hämagglutinin-Tag (HA-Tag, Sequenz: YPYDVPDYA)[10] ... ein C-terminales Protein-Tag am Protein während der Translation. Gelegentlich muss das Protein-Tag vom Protein nach der ... Zur Erzeugung eines Protein-Tags wird die codierende DNA-Sequenz des Protein-Tags unter Erhalt des Leserasters in die ... A generic protein purification method for protein complex characterization and proteome exploration. . In: Nature Biotechnology ...
Motor proteins then push the centrosomes along these microtubules to opposite sides of the cell. Although centrosomes help ... Semiopen orthomitosis occurs with different variants in some amoebae (Lobosa) and some green flagellates (e.g., Raphidophyta or ... Mitotic cells can be visualized microscopically by staining them with fluorescent antibodies and dyes. ... Volume 15 of Protein Reviews. Berlin: Springer Science & Business Media. p. 15. ISBN 9781461405146.. ...
Protein nanopore sequencing utilizes membrane protein complexes such as α-hemolysin, MspA (Mycobacterium smegmatis Porin A) or ... Ewing B, Green P (March 1998). "Base-calling of automated sequencer traces using phred. II. Error probabilities". Genome Res. 8 ... The fluorescent label is detached from the nucleotide upon its incorporation into the DNA strand, leaving an unmodified DNA ... a small protein secreted by the pancreas. This provided the first conclusive evidence that proteins were chemical entities with ...
"Grasse-green is made of Pink and Bice, it is shadowed with Indigo and Pink … French-green of Pink and Indico [shadowed with] ... Raw beef is red, because the muscles of vertebrate animals, such as cows and pigs, contain a protein called myoglobin, which ... The United States Manual on Uniform Traffic Control Devices specifies fluorescent pink as an optional color for traffic signs ... "Season Colour - I Think Spring is Green". Archived from the original on March 6, 2016. Retrieved February 17, ...
B) is the cyst wall selectively imaged through use of fluorescent-labelled (TRITC) antibody that is cyst wall specific.. (C) is ... Abdominal pain; diarrhea; vomiting; headache; yellow-green discolouration of urine Albendazole 5 ditë Dizziness; headache; ... 2003). "Mitochondrial remnant organelles of Giardia function in iron-sulphur protein maturation". Nature. 426 (6963): 172-6. ...
The thioredoxin system contains the 12-kDa protein thioredoxin and its companion thioredoxin reductase.[149] Proteins related ... Green AR, Ashwood T (April 2005). "Free radical trapping as a therapeutic approach to neuroprotection in stroke: experimental ... "Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe ... while damage to proteins causes enzyme inhibition, denaturation and protein degradation.[60] ...
Multiphoton fluorescence image of cultured HeLa cells with a fluorescent protein targeted to the Golgi apparatus (orange), ... HeLa cells grown in culture and stained with antibody to tubulin (green), antibody to Ki-67 (red) and the blue DNA binding dye ... Immunofluorescence image of HeLa cells grown in tissue culture and stained with antibody to actin in green, vimentin in red and ... Hou, S.Y.; Wu, S.; Chiang, C. (2002). "Transcriptional activity among high and low risk human papillomavirus E2 proteins ...
Susan Brooks: The discovery of aequorin and green fluorescent protein. In: Journal of Microscopy. Band 217, 2005, S. 1-2. ... Osamu Shimomura: The discovery of aequorin and green fluorescent protein. In: Journal of Microscopy. Band 217, 2005, S. 3-15. ... Hien huet 1961 d'Protein Aequorin an d'Protein dat gréng fluoreszéiert (GFP) an der Jelliskapp-Aart Aequorea victoria entdeckt ...
Organic solid waste (green waste)[edit]. A large compost pile that is steaming with the heat generated by thermophilic ... The larvae are rich in fat and protein, and can be used as, for example, animal feed or biodiesel production.[27] Enthusiasts ... At the simplest level, the process of composting requires making a heap of wet organic matter (also called green waste), such ... The USA is the only Western country that does not distinguish sludge-source compost from green-composts, and by default in the ...
2008 - Osamu Shimomura, Martin Chalfie, Roger Tsien for the discovery and development of the green fluorescent protein, GFP.[11 ... 2004 - Aaron Ciechanover, Avram Hershko, Irwin Rose for the discovery of ubiquitin-mediated protein degradation.[7] ... 2012 - Robert Lefkowitz and Brian Kobilka for studies of G-protein-coupled receptors.[14] ...
Green JP, Johnson CL, Weinstein H, Maayani S (December 1977). "Antagonism of histamine-activated adenylate cyclase in brain by ... β-arrestin over activating G proteins.[78][79] LSD also has an exceptionally long residence time when bound to serotonin ... LSD is strongly fluorescent and will glow bluish-white under UV light. ...
If the quantification of non-1,2-aminothiol-bearing protein is desired, the protein of interest can be cleaved to yield a ... K. Wang, X. Bi, S. Xing, P. Liao, Z. Fang, X. Meng, Q. Zhang, Q. Liu, Y. Ji Green Chem., 13 (2011), p. 562 ... Fluorescent azides and alkynes also produced by such companies as Active Motif Chromeon[60] and Cyandye ... SPAAC between a cyclooctyne-modified fluorophore and azide-tagged proteins allowed the selection of these proteins in cell ...
The green fluorescent protein (GFP) is often used in genetics as a marker. Many substances, such as proteins, have significant ... Fluorescent dye uses. Colorless fluorescent dyes that emit blue light under UV are added as optical brighteners to paper and ... Fluorescent black light lamps work similarly to other fluorescent lamps, but use a phosphor on the inner tube surface which ... UV fluorescent dyes are used in many applications (for example, biochemistry and forensics). Some brands of pepper spray will ...
Green sulfur bacteria (Chlorobium, Chloroherpeton). *Bacteroides, Flavobacteria and relatives (later renamed Bacteroidetes * ... gamma subdivision (enterics, fluorescent pseudomonads, purple sulfur bacteria, Legionella, (some) Beggiatoa). *delta ... Proteins are shown in blue and the single RNA strand in tan.[13] ... Green non-sulfur bacteria and relatives (later renamed ... Phylogenetic tree showing the diversity of bacteria, compared to other organisms.[1] Eukaryotes are colored red, archaea green ...
Stevens FJ, Solomon A, Schiffer M (1991). "Bence Jones proteins: a powerful tool for the fundamental study of protein chemistry ... Microtubules as shown in green, are marked by an antibody conjugated to a green fluorescing molecule, FITC. ... "New Dioxaborolane Chemistry Enables [18F]-Positron-Emitting, Fluorescent [18F]-Multimodality Biomolecule Generation from the ... and their realization that this protein is the same as the Bence-Jones protein described in 1845 by Henry Bence Jones.[101] ...
ProteinsEdit. The Mg2+ ion tends to bind only weakly to proteins (Ka ≤ 105[46]) and this can be exploited by the cell to switch ... By fluorescent indicatorsEdit. A number of chelators of divalent cations have different fluorescence spectra in the bound and ... Green vegetables such as spinach provide magnesium because of the abundance of chlorophyll molecules, which contain the ion. ... To date, only the ZntA protein of Paramecium has been shown to be a Mg2+ channel.[66] The mechanisms of Mg2+ transport by the ...
The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9 kDa) that exhibits bright green ... Wikimedia Commons has media related to Green fluorescent proteins.. *A comprehensive article on fluorescent proteins at ... Green Fluorescent Protein on FPbase, a fluorescent protein database. *Overview of all the structural information available in ... Excitation and emission spectra for various fluorescent proteins. *Green Fluorescent Protein Chem Soc Rev themed issue ...
... discovery and development of the green fluorescent protein (GFP), a naturally occurring substance in the jellyfish Aequorea ... Other articles where Green fluorescent protein is discussed: Martin Chalfie: … ... fluorescence of a protein called green fluorescent protein (GFP), which is excited through a chemical reaction (see below ... was working with variants of green fluorescent protein (GFP), a naturally occurring protein made by the jellyfish Aequorea ...
Researchers have pioneered a new way of controlling protein interactions, potentially opening the door to a host of medical and ... Green Fluorescent Protein. * Stanford researchers accidentally discover a whole new role for the cerebellum March 20, 2017 at ...
... Daniel J. Kliebenstein djk6 at CORNELL.EDU Mon Jul 25 16:36:10 EST 1994 *Previous message: parsley ... Howdy, I was wondering if anyone out there has been using the Green Fluorescent Protein described in Science by Chalfie, et al ... Also how much of the protein has to be present for the fluorescence to be detectable by the eye without any aids. I greatly ...
... From the University of California, Davis, Partnership for Plant ... coli to incorporate and express a plasmid containing a gene that codes for a green fluorescent protein (GFP). The source of the ... this biotechnology laboratory is a five-day activity with bacterial transformations and green fluorescent protein. "In this lab ... You just viewed Bacterial Transformation with Green.... Please take a moment to rate this material. ...
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 ...
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Berkeley researchers have identified the mechanism that makes green fluorescent protein (GFP) light up the way it does. They ... How Green Fluorescent Proteins Fluoresce. November 13th, 2009 Medgadget Editors News Berkeley researchers have identified the ... Berkeley Lab press release: Vibrations key to efficiency of green fluorescent protein …. Abstract in Nature: Mapping GFP ... mechanism that makes green fluorescent protein (GFP) light up the way it does. They used femtosecond lasers to image the ...
Flavin Mononucleotide-Based Fluorescent Reporter Proteins Outperform Green Fluorescent Protein-Like Proteins as Quantitative In ... The green fluorescent protein (GFP) of the jellyfish Aequorea victoria is an unusual protein with strong visible absorbance and ... Structural Dynamics of Green Fluorescent Protein Alone and Fused with a Single Chain Fv Protein ... Expression of Green Fluorescent Protein Fused to Magnetosome Proteins in Microaerophilic Magnetotactic Bacteria ...
In this work we have used for the first time green fluorescent protein (GFP) tagged cells of the human parasite Leishmania ... In this work we have used for the first time green fluorescent protein (GFP) tagged cells of the human parasite Leishmania ...
New fluorescent tool can track mysterious workings of cellular RNAs The ability to tag proteins with a green fluorescent light ... called green fluorescent protein (GFP), into the eggs from which the animals eventually grew. This method of genetic ... Crystal structures of key fluorescent proteins to light up molecules in cells Scientists at Albert Einstein College of ... Medicine of Yeshiva University have determined the crystal structures of two key fluorescent proteins - one blue, one red - ...
... a fluorescent molecule, found in the jellyfish Aequorea victoria, was the first of a diverse family of fluorescent proteins ... The green fluorescent protein from Aequorea victoria was the first intrinsically fluorescent protein to be discovered. ... detecting protein-protein interactions and protein expression patterns. In: Chalfie M and Kain S (eds) Green Fluorescent ... Green fluorescent protein (GFP), a fluorescent molecule, found in the jellyfish Aequorea victoria, was the first of a diverse ...
AcGFP1 is a monomeric green fluorescent protein and ZsGreen1 is extremely bright. ... Green Fluorescent Protein Vectors. Green fluorescent proteins provide a valuable, noninvasive approach for investigating ... ZsGreen1: An Extremely Bright Green Fluorescent Protein. ZsGreen1 is a bright green fluorescent protein that was derived from a ... Our Living Colors green fluorescent proteins (GFPs) include proteins suitable for fusion, cell labeling, and reporter studies; ...
Green-fluorescent protein fusions for efficient characterization of nuclear targeting.. Grebenok RJ1, Pierson E, Lambert GM, ... The green-fluorescent protein (GFP) from Aequorea victoria has been shown to be a convenient and flexible reporter molecule ... This implies that GFP might also be particularly suited for studies of intracellular protein targeting. In this paper, the use ... A novel oligopeptide motif from a tobacco protein is described which confers nuclear localization of GUS. The use of this ...
In addition see the different cases where this enhanced green fluorescent protein can transfer, as well as its efficiency. ... Enhanced Green Fluorescent Protein (EGFP) Chromophore. The GFP chromophore is encoded by the primary amino acid sequence, and ... so there has been limited use of this high-performance green variant. Emerald fluorescent protein contains the S65T, and F64L ... tutorial explores the molecular rearrangement that occurs during the formation of the enhanced green fluorescent protein (EGFP ...
GREEN FLUORESCENT PROTEIN. A. 237. Aequorea victoria. Mutation(s): 3 Gene Names: GFP. ... The structural basis for spectral variations in green fluorescent protein.. Palm, G.J., Zdanov, A., Gaitanaris, G.A., Stauber, ...
2004) Origin, nature, and fate of the fluorescent state of the green fluorescent protein chromophore at the CASPT2//CASSCF ... 2002) Green fluorescent protein (GFP): Applications, structure, and related photophysical behavior. Chem Rev 102(3):759-781. ... 1996) Chemical nature of the light emitter of the Aequorea green fluorescent protein. Proc Natl Acad Sci USA 93(24):13617-13622 ... 2001) Radiationless relaxation in a synthetic analogue of the green fluorescent protein chromophore. J Phys Chem B 105(33):8036 ...
Green Fluorescent Protein Helps Measure Intracellular Temperature. March 19th, 2012 Editors News ... Abstract in Nano Letters: Mapping Intracellular Temperature Using Green Fluorescent Protein. More from Nanowerk: A precise ... Spain have developed a method of using Green Fluorescent Protein (GFP) to do just that. GFP famously transformed bioscience ... Development on Recombinant Protein. • Biotech Companies and Market Analysis. Who Should Attend and Who Youll Meet?. Directors/ ...
Green fluorescent proteinImported. ,p>Information which has been imported from another database using automatic procedures.,/p ... View protein in InterPro. IPR009017 GFP. IPR011584 GFP-related. IPR000786 Green_fluorescent_prot. ... View protein in InterPro. IPR009017 GFP. IPR011584 GFP-related. IPR000786 Green_fluorescent_prot. ... tr,A7UAL5,A7UAL5_9CNID Green fluorescent protein OS=Montastraea cavernosa OX=63558 PE=2 SV=1 ...
Its gene was cloned in Escherichia colito express the tetrameric wild-type protein. The protein emits strong green... ... EosFP is a novel fluorescent protein from the stony coral Lobophyllia hemprichii. ... Tsien, R.Y.: The Green Fluorescent Protein, Annu. Rev. Biochem. 67 (1998), 509-544.CrossRefGoogle Scholar ... Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W. and Prasher, D.C.: Green Fluorescent Protein as a Marker for Gene Expression, ...
FACS-optimized mutants of the green fluorescent protein (GFP).. Cormack BP1, Valdivia RH, Falkow S. ... We have constructed a library in Escherichia coli of mutant gfp genes (encoding green fluorescent protein, GFP) expressed from ... In addition, when produced in E. coli, the folding of the mutant proteins is more efficient than folding of wt GFP. These two ... All three classes of mutant proteins have highly shifted excitation maxima. ...
Suppliers of: Green Fluorescent Protein (GFP). Found 13 Companies HOW TO USE BSN RAPID REQUEST. To send a request for quote/ ...
Green fluorescent protein-like proteinImported. ,p>Information which has been imported from another database using automatic ... tr,Q8T5E9,Q8T5E9_RICFL Green fluorescent protein-like protein OS=Ricordea florida OX=165100 PE=2 SV=1 ... to allow unambiguous identification of a protein.,p>,a href=/help/protein_names target=_top>More...,/a>,/p>Protein namesi. ... Integrated resource of protein families, domains and functional sites. More...InterProi. View protein in InterPro. IPR009017, ...
Green Fluorescent Protein Market To 2026 Seeking Excellent Growth , Novus Biologicals, COSMO BIO Co., Bio-Rad Laboratories - ... Green Fluorescent Protein Market Country Level Analysis:. The countries covered in the Green Fluorescent Protein Market report ... global-green-fluorescent-protein-market. Leading Green Fluorescent Protein manufacturers/companies operating at both regional ... Table Of Contents: Green Fluorescent ...
Michael Davidson Lab: Davidson Green fluorescent proteins Unpublished Plasmids from Article. <"clearfix">t data-page-length ...
Selection for RNAs that bind and activate derivatives of the green fluorescent protein fluorophore yields a wide range of ... Selection for RNAs that bind and activate derivatives of the green fluorescent protein fluorophore yields a wide range of ...
The chromophore of the green fluorescent protein, as a biomolecule from jellyfish Aequorea victoria, has been widely used as a ... M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, and D. C. Prasher, "Green fluorescent protein as a marker for gene expression," ... 21] found that cis-to-trans photoisomerization should be a general mechanism of green fluorescent protein chromophores whose ... Detailed Photoisomerization Dynamics of a Green Fluorescent Protein Chromophore Based Molecular Switch. Chen-Wei Jiang,1 Ai- ...
Researchers uncover the central role of a protein linked to Fragile X Syndrome in mice, one of the leading causes of autism and ... tags: green fluorescent protein x culture x The Scientist. » green fluorescent protein and culture ...
Researchers have used a modified rabies virus and fluorescent proteins to tag individual nerve cells in the mouse visual cortex ... Researchers uncover the central role of a protein linked to Fragile X Syndrome in mice, one of the leading causes of autism and ... tags: green fluorescent protein x immunology x The Scientist. » green fluorescent protein and immunology ...
A complementary DNA for the Aequorea victoria green fluorescent protein (GFP) produces a fluorescent product when expressed in ... Green fluorescent protein as a marker for gene expression Message Subject. (Your Name) has forwarded a page to you from Science ... GFP expression can be used to monitor gene expression and protein localization in living organisms. ...
  • This interactive tutorial explores the molecular rearrangement that occurs during the formation of the enhanced green fluorescent protein (EGFP) chromophore, which substitutes threonine for serine at position 65 in the amino acid sequence of the wild-type protein. (
  • The GFP-S65T variant was further modified by replacing phenylalanine for leucine at position 64 (F64L), which improved the efficiency of protein maturation at 37° C, yielding EGFP (enhanced GFP). (
  • Furthermore, EGFP is among the brightest and most photostable of the Aequorea-based fluorescent proteins. (
  • Continued engineering of EGFP has yielded several additional green variants with improved characteristics. (
  • Emerald fluorescent protein contains the S65T, and F64L mutations featured in EGFP, but also has four additional point mutations that further improve the efficiency of maturation and folding at 37° C, and increase the intrinsic brightness. (
  • We have been using the pEGFP-C3 and pEGFP-N3 vectors, which contain sequences encoding the enhanced version of green fluorescent protein (EGFP) whose expression is driven by the cytomegalovirus (CMV) promoter. (
  • Convenient restriction sites in these vectors allow fusion of a protein of interest to either the amino- or carboxy-terminus of EGFP. (
  • A fluorometric assay for pepsin and pepsinogen was developed using enhanced green fluorescent protein (EGFP) as a substrate. (
  • In this paper, we show that the AI-03 transgene that contains enhanced green fluorescent protein (EGFP) fused to the end of the neurophysin at the C-terminus of the OT pre-prohormone, is expressed selectively in OT-magnocellular neurons and is trafficked to secretory granules in transgenic mice. (
  • This antibody reacts with wild type Green fluorescent protein (GFP) from Aequorea victoria and its variants EGFP and EBFP. (
  • Results Using the same experimental procedures, we have compared Aequorea victoria enhanced green fluorescent protein (Av-eGFP) and Renilla raniformis GFP (Rh-GFP, h- from humanized) for the purpose of gene marking of hNSCs. (
  • We report the construction of two recombinant viruses, one having the enhanced green fluorescent protein (eGFP) fused in-frame to the pp38 open reading frame (ORF) (RB1Bpp38/eGFP) and the other having soluble-modified GFP (smGFP) downstream but out-of-frame with pp38 (RB1Bpp38/smGFP). (
  • The enhanced green fluorescent protein (eGFP) was efficiently applied as an optically active center. (
  • eGFP absorbs in the UV/visible range and converts it into green emission with a maximum absolute quantum yield of ∼0.50. (
  • Enhanced green fluorescent protein (EGFP) is a red shifted variant of wild-type GFP which has been optimized for brighter fluorescence and higher expression in mammalian systems. (
  • Cloning confirmation procedures and EGFP expression of transfected MCF-7 breast cancer cell line, additionally with hygromycin resistance phenotype of the green shining cells following treatment of the transfected cells with hygromycin B revealed that our newly synthesized hygromycin resistant EGFP fusion shuttle vector was constructed successfully. (
  • Expression of enhanced green fluorescent protein (eGFP) was constitutively driven in neurons using the herpes simplex virus amplicon system. (
  • [9] Frederick Tsuji's lab independently reported the expression of the recombinant protein one month later. (
  • Cloning of the cDNA and heterologous expression of fluorescent protein in a wide variety of organisms indicate that this unique posttranslational modification must be either spontaneous or dependent only on ubiquitous enzymes and reactants. (
  • The availability of two visibly distinct colors should significantly extend the usefulness of GFP in molecular and cell biology by enabling in vivo visualization of differential gene expression and protein localization and measurement of protein association by fluorescence resonance energy transfer. (
  • Fluorescent proteins are useful reporter molecules for in vitro and whole organism gene expression studies and for tracking subcellular localisation of proteins in living cells. (
  • 1998) New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. (
  • 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. (
  • In this work, we studied the use of green fluorescent protein (GFP) as a reporter for the magnetosomal localization and expression of fusion proteins in the microaerophilic Magnetospirillum gryphiswaldense by flow cytometry, fluorescence microscopy, and biochemical analysis. (
  • Under these optimized conditions, we studied the subcellular localization and expression of the GFP-tagged magnetosome proteins MamC, MamF, and MamG by fluorescence microscopy and immunoblotting. (
  • GFP has been used extensively as a fluorescent tag to monitor gene expression and protein localization. (
  • Green fluorescent protein as a marker for gene expression and cell biology of mycobacterial interactions with macrophages. (
  • We have taken a copy of this dimmer (promoter) and altered it such that it controls the expression of a green fluorescent protein in our cells. (
  • [3] Due to this stability, GFP could be applied to numerous other applications such as cell lineage tracing, gene expression reporting, or protein-protein interactions. (
  • Presumably, this is due to varying l evels of expression of exogenous proteins typical of transiently transfected cells. (
  • The increased transfection efficiency seen when using LipoTAXI transfection reagent creates a population of cells with various levels of exogenous protein expression. (
  • For some of our constructs, we have seen that expression level has a significant effect on the subcellular distribution of the exogenous protein (unpublished observations). (
  • Other effects of exogenous proteins, such as enzyme activity, may also be dependent on expression level. (
  • Use of LipoTAXI reagent provides a population of transfected cells containing diverse levels of expression of exogenous protein that may enhance studies. (
  • The green fluorescent protein (GFP) is a useful indicator in a broad range of applications including cell biology, gene expression and biosensing. (
  • The green fluorescent protein (GFP) of Aequorea victoria has been developed here as a reporter for gene expression and protein localization in Candida albicans. (
  • However, a major drawback is that protein expression can lead to the formation of insoluble aggregates. (
  • Protein expression was monitored through fluorescence intensity on a high-throughput format and SDS-PAGE 50000 45000 analysis. (
  • Green Fluorescent Protein permits a broad range of applications where it has functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. (
  • GFP can be fused to a wide variety of proteins of interest without significantly interfering with their assembly or function and its intrinsic fluorescent properties allow expression as fluorescent protein in virtual any organism. (
  • Aim of this research is to analyze properties of GFP by cloning, mutations, expression of proteins and purification. (
  • Objectives of this research are to sub-clone GFP into a vector and mutations are carried out by various mutagenesis experiments followed by expression of proteins and purification. (
  • Expression of the gene fusions in different sec mutants, as well as signal sequence processing assays, confirmed that the periplasmically localized hybrid proteins were exported by the sec -dependent pathway. (
  • While expression of hybrid proteins comprised of full-length MBP did not result in overproduction lethality characteristic of some exported β-galactosidase hybrid proteins, synthesis of shorter, exported hybrid proteins was toxic to the cells. (
  • 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 goal of this work was to construct recombinant MDVs having direct fusions of a marker gene, the green fluorescent protein (GFP), to pp38 in order to study the expression patterns and localization of this protein during stages of MDV infection. (
  • The different fluorescent protein coding sequences (genes) that can be easily excised using the flanking BsaI restriction sites and cloned into any other expression vector of choice. (
  • Our microplate readers answer your questions on nucleic acids: concentration, interaction with other nucleic acids or proteins, single nucleotide polymorphisms and expression of genes. (
  • Expression of a stable green fluorescent protein mutant in group B streptococcus. (
  • Green fluorescent protein (GFP) expression is a common labeling method for bacteria, enabling their identification in complex samples or monitoring of their subcellular locations in eukaryotic host cells. (
  • We have recently reported1 stable expression of a green fluorescent protein mutant (GFPmut3) with enhanced fluorescence intensity in GBS. (
  • 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. (
  • In contrast, under low intensities of blue light, the HF morph showed lower growth rates than the LF morph, indicating that trade-offs are associated with high levels of fluorescent protein expression under this condition. (
  • It is used primarily as a protein marker to test for expression of specific proteins. (
  • Cells expressing a subset of the GFP-cDNA expression library were screened to recover those in which the fluorescence signal diminished rapidly when protein synthesis was inhibited. (
  • Flow cytometric analysis of transgene expression in higher plants: green-fluorescent protein. (
  • Expression of green fluorescent protein (GFP) was observed in the callus and protocorm-like body (PLBs) tissues survived on the selection medium. (
  • 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 ). (
  • The wild-type gfp gene 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. (
  • Expression of scFv-GFP fusion proteins using bacteria has resulted in limited success, with yields of 100 to 200 μg/liter, whether periplasmic secretion or inclusion body methods were used ( 2 , 8 , 18 , 20 , 21 ). (
  • Like mammalian and insect hosts, yeast is an effective eukaryotic expression host with protein quality control machinery helping to ensure that secreted protein is active, although this filter is not perfect ( 16 ). (
  • While the MP acts like a microtubule-associated protein able to stabilize microtubules during late infection stages, the protein was also shown to cause the inactivation of the centrosome upon expression in mammalian cells, thus suggesting that MP may interact with factors involved in microtubule attachment, nucleation, or polymerization. (
  • However, efficient transfection is needed for expression of recombinant protein in mammalian cell. (
  • This localization was obtained by transforming strain RB51 with plasmids pBBg18sGFP and pBBgSsGFP, in which the 18 kDa Brucella lipoprotein and the Brucella Cu/Zn SOD protein signal sequences were added to the GFP sequence to cause OM and PS expression respectively. (
  • Green florescent Protein (GFP), originally isolated from the jellyfish Aequorea Victoria, has proven to be a useful reporter for monitoring gene expression and protein localization in vivo and in real time when fused to an intracellular or secretary protein. (
  • When irradiated with UV light or blue light, it emits green light, which enables the examination of expressed gene after expression. (
  • Synthetic green fluorescent protein (GFP) was used as a reporter to detect differential gene expression in the pathogenic fungus Cryptococcus neoformans . (
  • Expression of GFP5 results in greatly improved levels of fluorescence in both microbial and mammalian cells cultured at 37°C. Conclusions The thermotolerant mutants of GFP greatly improve the sensitivity of the protein as a visible reporter molecule in bacterial, yeast and mammalian cells. (
  • Construction of a transcriptional fusion between actA-plcB and the green fluorescent protein gene of Aequorea victoria has facilitated the detailed examination of patterns of actA/plcB expression within infected tissue culture cells. (
  • We stably induced the expression of human PG and GFP -Green Fluorescent Protein- as control in 3T3L1 cells using a lentiviral system to study the effect of PG expression in the differentiation capacity of this cell line, one of the most used adipogenic models. (
  • PG expression in 3T3L1 cells promotes changes in several Biological Processes, including structure of cytoskeleton, lipid metabolism, calcium regulation, translation, protein folding and energy generation by the mitochondria. (
  • Abstract: Experimental tools to determine membrane topology of a protein are rather limited in higher eukaryotic organisms. (
  • Structure of the Aequorea victoria green fluorescent protein. (
  • [2] [3] Similar proteins that also fluoresce green are found in many marine organisms, but the label GFP traditionally refers to this particular protein, which was first isolated from the jellyfish Aequorea victoria and is sometimes called-when such precision is required- avGFP . (
  • In the 1960s and 1970s, GFP, along with the separate luminescent protein aequorin (an enzyme that catalyzes the breakdown of luciferin , releasing light), was first purified from the jellyfish Aequorea victoria and its properties studied by Osamu Shimomura . (
  • discovery and development of the green fluorescent protein (GFP), a naturally occurring substance in the jellyfish Aequorea victoria that is used as a tool to make visible the actions of certain cells. (
  • was working with variants of green fluorescent protein (GFP), a naturally occurring protein made by the jellyfish Aequorea victoria . (
  • The green fluorescent protein (GFP) of the jellyfish Aequorea victoria is an unusual protein with strong visible absorbance and fluorescence from a p-hydroxybenzylidene-imidazolidinone chromophore, which is generated by cyclization and oxidation of the protein's own Ser-Tyr-Gly sequence at positions 65-67. (
  • Green fluorescent protein (GFP), a fluorescent molecule, found in the jellyfish Aequorea victoria , was the first of a diverse family of fluorescent proteins cloned from marine invertebrates. (
  • The green fluorescent protein from Aequorea victoria was the first intrinsically fluorescent protein to be discovered. (
  • Proteins pictured are variants of Aequorea victoria GFP and Discosoma sp. (
  • The label GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria and is sometimes called avGFP. (
  • The chromophore of the green fluorescent protein, as a biomolecule from jellyfish Aequorea victoria , has been widely used as a genetically encoded noninvasive fluorescence marker in bioimaging [ 20 ]. (
  • 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. (
  • Picosecond time-resolved mid-infrared absorption changes of the wild type green fluorescent protein from Aequorea victoria are reported on structural events during the photocycle. (
  • B JO - J Phys Chem B VL - 109 IS - 33 N2 - Picosecond time-resolved mid-infrared absorption changes of the wild type green fluorescent protein from Aequorea victoria are reported on structural events during the photocycle. (
  • The green fluorescent protein ( GFP ) is a protein , comprised of 238 amino acids (26,9 kDa ), from the jellyfish Aequorea victoria that fluoresces green when exposed to blue light. (
  • Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria . (
  • Green fluorescence protein ( GFP ) is a 27 kDa protein derived from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelenth of 509 nm) when excited by blue light (excitation peak at a wavelenth of 395 nm). (
  • a protein obtained from the jellyfish Aequorea victoria that emits a bright green fluorescence when illuminated. (
  • Discovered only in the mid-1990s, green fluorescent protein (GFP) from the jellyfish Aequorea victoria has become a workhorse of molecular biology. (
  • rGFP, also known as Green Fluorescent Protein, is a protein produced by the jellyfish (Aequorea Victoria) that produces bioluminescence in the green zone of the noticeable spectrum. (
  • These green fluorescent proteins from other organisms have different sequences but they resemble the GFP from Aequorea victoria in their molecular structure and properties. (
  • Background -- Fluorescent proteins (FP) homologous to the green fluorescent protein (GFP) from the jellyfish Aequorea victoria have revolutionized biomedical research due to their usefulness as genetically encoded fluorescent labels. (
  • Even though many other marine organisms have comparable green fluorescent proteins, GFP is generally referred as the first isolated protein from jellyfish and Aequorea Victoria so called crystal jelly also discovers green fluorescent protein. (
  • Haseloff, Jim 1996-12-01 00:00:00 Background The green fluorescent protein (GFP) of the jellyfish Aequorea victoria has recently attracted great interest as the first example of a cloned reporter protein that is intrinsically fluorescent. (
  • Yellow fluorescent protein (YFP) is a genetic mutant of green fluorescent protein (GFP) originally derived from the jellyfish Aequorea victoria. (
  • Four protein mutations from the wild-type GFP found in Aequorea Victoria jellyfish was needed to create the YFP mutant. (
  • AcGFP1 has been widely validated as a fusion tag, using a wide variety of proteins with diverse functions and subcellular locations (Figure 1, Panel C). AcGFP1 is particularly suited for use in multicolor applications, e.g., to simultaneously visualize the subcellular localization of two proteins of interest. (
  • In summary, our data demonstrate the use of GFP as a reporter for protein localization under magnetite-forming conditions and the utility of MamC as an anchor for magnetosome-specific display of heterologous gene fusions. (
  • Second, we have noticed heterogeneous subcellular localization of some of our fusion proteins in the same set of transfected cells. (
  • The use of green fluorescent protein (GFP) as a reporter for protein localization in Escherichia coli was explored by creating gene fusions between malE (encoding maltose-binding protein [MBP]) and a variant of gfp optimized for fluorescence in bacteria (GFPuv). (
  • Haseloff, J., Siemering, K.R., Prasher, D.C., Hodge, S.: Removal of a crytic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. (
  • The genome of Arabidopsis ( Arabidopsis thaliana ) contains seven VSR proteins, but little is known about their individual subcellular localization and function. (
  • Here, we study the subcellular localization of the seven Arabidopsis VSR proteins (AtVSR1-7) based on the previously proven hypothesis that the TMD and CT sequences correctly target individual VSR to its final destination in transgenic tobacco BY-2 cells. (
  • In order to determine if the immune response is affected by the localization of the antigen, green fluorescent protein (GFP) was expressed at three different locations in B. abortus strain RB51, outer-membrane (OM), periplasmic space (PS) and in the cytoplasmic region (CR) of B. abortus strain RB51. (
  • Spectrally resolved imaging of Green fluorescent protein (GFP) expressed in living COS-7 kidney cells distinguished the subcellular localization and demarcated the processes of protein folding and chromophore formation. (
  • The genes for both proteins have been human codon-optimized to enhance their translation in mammalian cells (2), and the proteins have been optimized for bright emission and fast chromophore maturation. (
  • Fluorescent proteins are highly useful reporter genes in a wide variety of biological systems. (
  • The gene coding for it can be tagged to the genes of other proteins or viruses to study their movements within cells. (
  • The study of bacterial protein export has been greatly facilitated by the use of reporter genes whose products serve as enzymatic markers for cellular location. (
  • Many of these mutants ultimately led to the identification of a number of sec genes that encode important components of the Escherichia coli protein export machinery ( 17 , 30 , 40 ). (
  • This "marking" makes it possible to understand whether genes work or not, and whether they are active in protein production or not. (
  • The presence of green fluorescence protein (s gfp ), hygromycin-B-phosphotransferase ( hptII ) and β-glucuronidase ( uidA ) genes in the transformed tissues were verified using PCR, Southern blot and dot blot analyses. (
  • The laser snapshots show that when the light absorber, or chromophore, nestled in the middle of the protein barrel absorbs an incoming photon of blue light, it starts vibrating, and the electrons start sloshing around the chromophore until it is aligned just right for the proton to hop via a water molecule to a nearby amino acid in the protein. (
  • From there, it continues down the reaction chain, creating a state with a negatively charged chromophore that emits green light. (
  • Previous studies had shown that after the chromophore absorbs blue light, it undergoes proton transfer, and green light is emitted. (
  • However, the wagging oscillation might have stopped after a few picoseconds, when the chromophore and its vicinity are aligned just right for the proton to hop off down the reaction chain, and the whole protein shines bright green - which it does in its own good time, in about 3 nanoseconds. (
  • ZsGreen1 is a bright green fluorescent protein that was derived from a reef coral belonging to class Anthozoa (1), and has been modified for higher solubility, brighter emission, and rapid chromophore maturation (8-12 hours) compared to the unmodified protein. (
  • By elucidating an energetics-function model for split GFP strand photodissociation, we show that photodissociation is achieved by light-activated cis-trans isomerization of the chromophore, a mechanism pervasive among reversibly photoswitchable fluorescent proteins. (
  • Ai H‐W, Shaner NC, Cheng Z, Tsien RY and Campbell RE (2007) Exploration of new chromophore structures leads to the identification of improved blue fluorescent proteins. (
  • The GFP chromophore is encoded by the primary amino acid sequence, and forms spontaneously without the requirement for cofactors or external enzyme components (other than molecular oxygen), through a self-catalyzed protein folding mechanism and intramolecular rearrangement. (
  • The protein emits strong green fluorescence (516 nm) that shifts toward red (581 nm) upon near-ultraviolet irradiation at ∼390 nm due to a photo-induced modification that involves a break in the peptide backbone next to the chromophore. (
  • Recently, several studies have revealed that the green fluorescent protein chromophore and its different modifications undergo photo-induced Z/E isomerization under radiation [ 21 - 26 ], which offer them the opportunity to be molecular switches. (
  • Through investigation of several spectral modification of synthetic chromophore analogues of wild-type green fluorescent protein, Voliani et al. (
  • 22 ] investigated two green fluorescent protein chromophore analogs and proposed a multicoordinate relaxation mechanism. (
  • Very recently, several oxazolone analogs of the green fluorescent protein chromophore were synthesized and predicted to be good candidates for molecular switches by Blanco-Lomas et al. (
  • To furnish a deeper and detailed mechanistic understanding of the photoisomerization reaction of those green fluorescent protein chromophore based molecular switches, semiclassical nonadiabatic molecular dynamics simulations were performed for a molecule 4-benzylidene-2-methyloxazol-5(4H)-one (BMH, named as 2e in [ 27 ]) in our group. (
  • Green links depict the chromophore of each GFP variant. (
  • Protein and chromophore folding also constitutes as a major advantage of GFP. (
  • The neutral form of the chromophore in wild-type green fluorescent protein (wtGFP) undergoes excited-state proton transfer (ESPT) upon excitation, resulting in characteristic green (508 nm) fluorescence. (
  • This ESPT reaction involves a proton relay from the phenol hydroxyl of the chromophore to the ionized side chain of E222, and results in formation of the anionic chromophore in a protein environment optimized for the neutral species (the I* state). (
  • Reorientation or replacement of E222, as occurs in the S65T and E222Q GFP mutants, disables the ESPT reaction and results in loss of green emission following excitation of the neutral chromophore. (
  • An increasing number of non-natural amino acids are available for chromophore redesign (by engineering of the genetic code) and enable new general strategies to generate novel classes of tailor-made GFP proteins. (
  • The typical GFP spectral fingerprint is the result of protein folding and chromophore formation following internal oxidation reactions. (
  • A series of studies quantified the weak interactions between the chromophore and the protein environment and enable us to explain the properties of GFP. (
  • It also performs well in cell-based assays that monitor protein subcellular trafficking (Figure 1, Panels A and B). Cells expressing AcGFP1 are easily detected and sorted by flow cytometry. (
  • Optical techniques for investigating biological processes in vivo can achieve subcellular spatial and millisecond temporal resolution by using genetically encoded light-responsive proteins ( 1 ). (
  • Additionally, photoswitchable or photoconvertible proteins can be employed in novel microscopy concepts that allow imaging of subcellular structures at a resolution beyond the diffraction barrier of optical microscopy [22] - [24] . (
  • Thus, genetically-encoded GFP provided for the first time the ability to label specific proteins inside the living cell without the need for exogenous synthetic or antibody-labeled fluorescent tags. (
  • Mild and cost-effective green fluorescent protein purification employing small synthetic ligands. (
  • Recombinant synthetic non-Aequorea Green Fluorescent Protein was expressed in E. coli. (
  • Synthetic non-Aequorea fluorescent proteins can bring a world of color to your research. (
  • rGFP is expressed in most known cell types and is used as a noninvasive fluorescent marker in living cells and organisms. (
  • [10] Remarkably, the GFP molecule folded and was fluorescent at room temperature, without the need for exogenous cofactors specific to the jellyfish. (
  • Femtosecond stimulated Raman spectroscopy on GFP involves hitting the protein molecule with an approximately 80 femtosecond pulse of ultraviolet light, which excites many vibrational modes in the molecule, and then a one-two punch of picosecond red and femtosecond white light to stimulate Raman emission. (
  • These studies demonstrate that introducing GFP as a fusion within the context of a rapidly degraded protein does not alter the degradation properties of the parent molecule, and that the GFP moiety of the fusion protein is degraded along with the rest of the protein. (
  • This has triggered the development of highly automated live cell fluorescence microscopy systems which can be used to observe cells over time expressing one or more proteins tagged with fluorescent proteins. (
  • Host colonization and virulence are mainly studied using microscopy and fluorescent biomarkers. (
  • The accessibility of GFP and its derivatives has meticulously interpreted fluorescent microscopy and the technique worn in cell ecology and other natural disciplines. (
  • We quantified biomass of a specific fungal biological control agent in nonsterile soil using epifluorescence microscopy and image analysis of green fluorescent protein (GFP)-expressing Trichoderma harzianum (ThzID1-M3). (
  • Bsr can be detected by fluorescent microscopy or flow cytometry. (
  • As with all of our fluorescent proteins, AcGFP1 is well tolerated by mammalian cells, and has been successfully used to establish stably transfected, clonal cell lines. (
  • Our lab has recently been interested in the transient transfection of green fluorescent proteins into mammalian cells. (
  • By using mammalian gGFP, the membrane topology of disease-associated membrane proteins, URG7, MRP6{sub 102}, SP-C(Val) and SP-C(Leu) was confirmed. (
  • The libraries were thus deemed to be useful for screening short-lived proteins in mammalian cells. (
  • In addition to bacterial systems, relatively efficient secretion of scFv-GFP fusion proteins at up to 50 to 3,000 μg/liter from transiently transfected mammalian cells and insect cells has been achieved ( 24 ). (
  • This observation suggests that these proteins can be engineered to provide active control of in vivo processes as optogenetic tools in addition to their conventional roles as passive imaging reporters. (
  • The mutant protein in fluorinated form showed improved folding efficiency both in vivo and in vitro, and the median fluorescence of cells expressing the fluorinated protein was improved {approx}650-fold in comparison to that of cells expressing fluorinated GFPm. (
  • These proteins may therefore become valuable additions to the in vivo imaging toolkit. (
  • In vitro assay of degradation is simpler than in vivo analysis, but an in vitro assay system may not fully mimic the degradation of proteins in the cells. (
  • The two proteins colocalize and interact in vivo as well as in vitro and exhibit mutual functional interference. (
  • This vector can be used as an easily detectable reporter and/or protein fusion vector for a number of basic experimental applications in situ and in vivo. (
  • Another phenotype associated with E. coli strains that produce exported β-galactosidase fusions is overproduction lethality, resulting from high-level synthesis of the hybrid proteins ( 3 , 13 ). (
  • Transgenic tobacco BY-2 cell lines expressing these seven sp-GFP-TMD-CT fusions all exhibited typical punctate signals colocalizing with VSR proteins by confocal immunofluorescence. (
  • In recent years, GFP-like proteins from marine invertebrates have emerged as indispensable tools for tracking of proteins or cells, sensing of intracellular conditions such as Ca 2+ or pH variations and for studies of protein interactions and gene activity, opening several stimulating perspectives in live cell imaging [1] - [9] . (
  • Activation of a GPCR results in the modulation of intracellular second messenger levels and/or ionic conductances via the coupling of receptors to a wide variety of effector systems via heterotrimeric guanine nucleotide-binding proteins (G proteins). (
  • By evaluating leader sequence processing and using GFP fluorescence to track intracellular processing, it was determined that the majority of fusion protein synthesized by the yeast was not secreted and in most cases was accumulating in an immature, although active, endoplasmic-reticulum (ER)-processed form. (
  • The results indicated that yeast can be used as an effective host for the secretion of scFv/scTCR-GFP fusion proteins and that as a result of intracellular secretory bottlenecks, there is considerable yeast secretory capacity remaining to be exploited. (
  • In cardiac precursor cells, I Ca was found to be already under control of cAMP-dependent phosphorylation since intracellular infusion of the catalytic subunit of protein kinase A resulted in a 1.7-fold stimulation. (
  • The β 2 -adrenergic receptor (β 2 AR) is prototypic of the large family of G protein-coupled receptors (GPCRs) whose desensitization and resensitization are regulated by intracellular kinases, arrestin proteins, phosphatases, and ill-defined components of the cellular endocytic machinery. (
  • The ActA protein of Listeria monocytogenes is an essential virulence factor and is required for intracellular bacterial motility and cell-to-cell spread. (
  • Ai H‐W, Henderson JN, Remington SJ and Campbell RE (2006) Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein: structural characterization and applications in fluorescence imaging. (
  • Characterization of green fluorescent protein (GFP)-expressing and progerin (PG)-expressing 3T3L1 cells. (
  • For example, the Shaker fusion protein mentioned in the section above was the first genetically encoded optical sensor of membrane potential that induced at most a 5% decrease in fluorescence in phosphorylated conditions. (
  • The main aim of this work was to study the impact of different temperatures after induction (27, 30, 33 and 37ºC) and IPTG levels (0.1, 0.5 and 1 mM) on the overall productivity of Green Fluorescent Protein (GFP) fusion proteins and on the ratio between soluble GFP fusion protein and IB formation. (
  • The fusion protein used on these studies was a GFP fused at the N - terminal to a hydrophobic peptide. (
  • As a model system, E. coli BL21(DE3) with pET 21C plasmid containing the gene encoding for the GFP fusion protein have been used. (
  • Western blot analysis of whole cell lysates from cells infected with adenovirus coding for GFP (A and C) or GFP-PTG (protein-targeting to glycogen) fusion protein. (
  • The periplasmic location of the fusion protein was confirmed by the observation that osmotic shock greatly decreased the periplasmic fluorescence signal by loss of the protein but had no effect on the fluorescence of the cytoplasmic protein. (
  • This fusion protein absorbs blue light (major peak at 480 nm) and emits green light (major peak at 505 nm). (
  • Fusion protein secretion levels ranged over 3 orders of magnitude, from 4 μg/liter to 4 mg/liter, and correlated well with the secretion levels of the unfused scFv/scTCR. (
  • Although the fusion protein secretion levels were not significantly affected by the nature of the fusion construct, the properties of the fusion protein were clearly influenced. (
  • In terms of the antibody subunit of a GFP fusion protein, single-chain antibodies (scFvs) consisting of the variable heavy- and light-chain regions of intact antibodies retain binding activity and can be efficiently processed by microbial production hosts. (
  • The key to an effective scFv-GFP fusion protein platform is the ability to rapidly take an identified scFv or collection of scFvs and produce them efficiently as GFP fusion proteins in an appropriate host. (
  • Using this method, we have separated GFP and GFP-insulin-like growth factor-I (GFP-IGF-I) fusion protein. (
  • AcGFP1 is an engineered, fluorescent mutant of the wild-type protein derived from the jellyfish Aequorea coerulescens . (
  • Scientists often link GFP to other specific proteins, and GFP reveals their location when it fluoresces. (
  • U.S. researcher Eric Poeschla has produced three glowing genetically modified cats by using a virus to carry a gene, called green fluorescent protein (GFP), into the eggs from which the animals eventually grew. (
  • Fluorescent proteins have been the subject of heavy engineering efforts to improve their physical and optical properties as genetically encoded tools. (
  • A colorful variety of fluorescent proteins (FPs) from marine invertebrates are utilized as genetically encoded markers for live cell imaging. (
  • Tranfection is a method that randomly introduce foreign DNA into cells either to produce genetically modified cells or to produce recombinant protein. (
  • Typically, YFP serves as the acceptor for genetically-encoded FRET sensors of which the most likely donor FP is monomeric cyan fluorescent protein (mCFP). (
  • Its gene was cloned in Escherichia coli to express the tetrameric wild-type protein. (
  • Cytoplasmic pH and periplasmic pH of Escherichia coli cells in suspension were observed with 4-s time resolution using fluorimetry of TorA-green fluorescent protein mutant 3* (TorA-GFPmut3*) and TetR-yellow fluorescent protein. (
  • Molecular biology and mutation of green fluorescent protein. (
  • Fluorescence (indicated by a green glow surrounding the affected structural elements) occurs when oxidation of the tyrosine alpha-beta carbon bond by molecular oxygen extends electron conjugation of the imidazoline ring system to include the tyrosine phenyl ring and its para-oxygen substituent (termed a para-hydroxybenzilidine moiety). (
  • Green fluorescent protein (GFP), molecular model. (
  • Therefore, our findings disclose the deep sea as a potential source of GFP-like molecular marker proteins. (
  • These green fluorescent proteins are now widely used in molecular biology and biochemistry. (
  • 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. (
  • Like the parent GFP, YFP is a useful tool in cell and molecular biology because the excitation and emission peaks of YFP are distinguishable from GFP which allows for the study of multiple processes/proteins within the same experiment. (
  • It has yet remained unknown if species expressing green fluorescent protein (GFP)-like proteins also exist in the darkness of the deep sea. (
  • Proteins from all species that are tagged with GFP. (
  • The application of FPs derived from marine species has been further extended by protein engineering. (
  • 00) provides the first popular science book on green fluorescent protein , which has existed in one species of jellyfish and was cloned recently to provide a host of new biotech applications. (
  • Scientist Dr. Chun-Jung Chen and Research Assistant Mr. Yen-Chieh Huang of the National Synchrotron Radiation Research Center collaborated with the researchers at the University of the Philippines - Diliman to analyze the three-dimensional structure and functional characteristics of the novel green fluorescent protein asFP504 isolated from a soft coral species, Alcyonium sp. (
  • Different tissue concentrations of these GFP-like proteins distinguish color morphs that are characteristic for many coral species. (
  • Green fluorescent protein (GFP) fluorescence was not detected in the third species, Pseudoalteromonas sp. (
  • As a step toward expanding understanding, we took a relatively unique approach by concentrating on a set of SG neurons selectively labeled by green fluorescent protein (GFP) in a transgenic mouse. (
  • Lessons from transgenic zebrafish expressing the green fluorescent protein (GFP) in the myeloid lineage. (
  • Selection for RNAs that bind and activate derivatives of the green fluorescent protein fluorophore yields a wide range of useful spectral properties. (
  • However, the development of marker proteins with entirely novel spectral properties greatly relies on the discovery of natural lead structures. (
  • These properties make GFP an ideal fluorescent marker for a broad range of biological applications and have sparked the development of variant fluorescent proteins with different spectral properties to allow simultaneous multi-colour reporter experiments. (
  • Further mutagenesis experiments are carried out by designing oligonucleotide primers which will alter the spectral properties of the protein. (
  • The gene sequence for GFP can be inserted into the DNA of an organism and thereby confer a new property, the ability to emit green fluorescence. (
  • p>This section provides information about the protein and gene name(s) and synonym(s) and about the organism that is the source of the protein sequence. (
  • section shows the unique identifier assigned by the NCBI to the source organism of the protein. (
  • GFP is fused in frame with the gene encoding the protein of interest, resulting in a chimera that is both functional (hopefully) and fluorescent to be expressed in the organism. (
  • While most small fluorescent molecules such as fluorescent isothiocyanate are powerful in formation of oxygen radicals due to non-radioactive energy transfer when used in live cells, fluorescent proteins such as GFP are typically much less injurious when injected in living organism. (
  • The resulting GFP-S65T mutant was a distinct improvement over wild-type GFP (wtGFP) for applications as a fluorescent marker in living cells because it had a well-defined absorption profile with a single peak at 489 nanometers. (
  • Although this near-wtGFP was fluorescent, it had several drawbacks, including dual peaked excitation spectra, pH sensitivity, chloride sensitivity, poor fluorescence quantum yield, poor photostability and poor folding at 37 °C. The first reported crystal structure of a GFP was that of the S65T mutant by the Remington group in Science in 1996. (
  • 21 ] found that cis -to- trans photoisomerization should be a general mechanism of green fluorescent protein chromophores whose efficiency can be modulated by the detailed mutant-specific protein environment. (
  • A similar recovery of green fluorescence is also observed for the E222Q/H148D mutant, suggesting that D148 is the proton acceptor for the ESPT reaction in both double mutants. (
  • Results We have carried out a screen for mutant forms of GFP that fluoresce more intensely than the wild-type protein when expressed in E. coli at 37°C. We have characterized a bright mutant (GFPA) with reduced sensitivity to temperature in both bacteria and yeast, and have shown that the amino acids substituted in GFPA act by preventing temperature-dependent misfolding of the GFP apoprotein. (
  • Nuclear accumulation of a mutant form of the nuclear protein Lamin-A, called Progerin (PG) or Lamin AΔ50, occurs in Hutchinson-Gilford Progeria Syndrome (HGPS) or Progeria, an accelerated aging disease. (
  • Scientists at Albert Einstein College of Medicine of Yeshiva University have determined the crystal structures of two key fluorescent proteins - one blue, one red - used to "light up" molecules in cells. (
  • Advanced uses for fluorescent proteins include optical sensing of important signalling molecules such as calcium and biochemical activities such as kinase activation. (
  • [6] While most small fluorescent molecules such as FITC (fluorescein isothiocyanate) are strongly phototoxic when used in live cells, fluorescent proteins such as GFP are usually much less harmful when illuminated in living cells. (
  • Förster resonance energy transfer (FRET) is a laboratory technique used to detect the proximity of two biological molecules by the fusion of fluorescent proteins with each of the proteins of interest. (
  • These proteins, which are complex structured giant molecules, have a very special geometry and pattern. (
  • However, few reports are available on the analysis in protein molecules. (
  • Photoactivatable proteins are used as either imaging tools, such as reversibly photoswitchable fluorescent proteins (RSFPs), with fluorescence that can be modulated by light ( 2 ) or optogenetic switches that convert light input into physiological outputs, such as channelrhodopsins, which can regulate ion flow through membranes in response to light ( 3 ). (
  • Photoactivatable and photoswitchable fluorescent proteins can be used for 'superresolution' imaging of cellular structures below the optical diffraction limit. (
  • Bioluminescent creatures live with special proteins that absorb light energy. (
  • Unlike other bioluminescent reporters, GFP requires no additional proteins, substrates or co-factors to emit light. (
  • We show that a herpes virus vector expressing a bioluminescent protein allows detailed morphometric analyses of living neurons in complex culture environments. (
  • Although this near-wtGFP was fluorescent, it had several drawbacks, including dual peaked excitation spectra, pH sensitivity, chloride sensitivity, poor fluorescence quantum yield, poor photostability and poor folding at 37 °C. (
  • Although this wild-type GFP was fluorescent, it had several drawbacks, including dual peaked excitation spectra, poor photostability and poor folding at 37°C. (
  • GFP (green fluorescent protein) is a protein of about 27kDa that emits green fluorescent light (emission maximum at 509 nm) when excited by blue or ultraviolet light (excitation maximum at 395 nm). (
  • This protein has Excitation of 400nm and Emission of 515nm. (
  • Then, the intensity of Green Fluorescent Protein (GFP) was quantified using fluorescence reader at emission wavelength 506 nm and excitation wavelength 500 nm. (
  • The diversity of genetic mutations is illustrated by this San Diego beach scene drawn with living bacteria expressing 8 different colors of fluorescent proteins (derived from GFP and dsRed). (
  • Fitness cost of the green fluorescent protein in gastrointestinal bacteria. (
  • The magnetosomes of magnetotactic bacteria are prokaryotic organelles consisting of a magnetite crystal bounded by a phospholipid bilayer that contains a distinct set of proteins with various functions. (
  • Students isolate the green fluorescent protein (GFP) and learn about recombinant proteins and genetic engineering along the way. (
  • The distinction between fluorescent and nonfluorescent colonies was exploited as a scorable phenotype to isolate malE signal sequence mutations. (
  • Students also learn about how genetic engineers use the properties of designer proteins to isolate them from the complex mixture of bacterial proteins and ultimately apply them to address human needs. (
  • But in the 1960s, scientists managed to isolate green fluorescent proteins. (
  • 2002) A monomeric red fluorescent protein. (
  • GFP is a 27 kDa monomeric protein, which autocatalytically forms a fluorescent pigment. (
  • The wild type protein absorbs blue light (maximally at 395nm) and emits green light (peak emission 508nm) in the absence of additional proteins, substrates, or co-factors. (
  • Addgene: Fluorescence resonance energy transfer using color variants of green fluorescent protein. (
  • In order to address this demand, a novel adsorbent was developed as a generic platform for the purification of GFP or GFP fusion proteins, giving GFP a dual function as reporter and purification tag. (
  • Purification of MBP-GFP hybrid protein from the different cellular compartments indicated that GFP is improperly folded when localized outside of the cytoplasm. (
  • 3D reconstruction of confocal image of VEGF-overexpressing neural progenitors (red) and GFP-positive control neural progenitor cells (green) in the rat olfactory bulb. (
  • Green fluorescent proteins provide a valuable, noninvasive approach for investigating biological events in living cells and tissues. (
  • Like all of Clontech's fluorescent proteins, AcGFP1 and ZsGreen1 can be detected in cells without adding cofactors or substrates, which makes them ideal for use in live cell assays (1). (
  • In this work we have used for the first time green fluorescent protein (GFP) tagged cells of the human parasite Leishmania donovani to observe its development in the gut of phlebotomine sand flies. (
  • The ability to tag proteins with a green fluorescent light to watch how they behave inside cells so revolutionized the understanding of protein biology that it earned the scientific teams who developed the technique Nobel Prizes in 2008. (
  • Colour variants of fluorescent proteins allow multicolour protein tracking in living cells. (
  • Reliably measuring the temperature of the inside of cells has been difficult to accomplish, but now researchers at The Institute of Photonic Sciences in Catalonia, Spain have developed a method of using Green Fluorescent Protein (GFP) to do just that. (
  • Researchers have used a modified rabies virus and fluorescent proteins to tag individual nerve cells in the mouse visual cortex. (
  • Another powerful use of GFP is to express the protein in small sets of specific cells. (
  • The lab of Martin Chalfie expressed the coding sequence of fluorescent GFP in heterologous cells of E. coli and C. elegans , publishing the results in Science in 1994. (
  • MedicalXpress)-Researchers from Ohio State University have designed nanoparticles based on naturally occurring green fluorescent proteins that can trace the progression of a cancer drug through human cells. (
  • Use green fluorescent protein, or GFP, and the cells will light up. (
  • Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells ( Comparison of liposome-based transfect. (
  • Here we compare the transfection efficiency of green fluorescent protein into human adrenalcarcinoma cells using both LipoTAXI transfection reagent and LipofectAMINE reagent. (
  • Transfection efficiency was determined by counting fluorescent and total cells from six random fields for each condition. (
  • Figure 1 shows fluorescent images of cells transfected under optimal conditions for either LipofectAMINE or LipoTAXI transfection reagents. (
  • C. albicans cells expressing this construct contained GFP mRNA but were non-fluorescent and contained no detectable translation product. (
  • C. albicans cells expressing this yeast-enhanced GFP gene (yEGFP3) are fluorescent and contain GFP protein. (
  • Flow cytometric analysis of cells expressing proteins in media supplemented with 1 mM Tfl and 45 mM Leu. (
  • The fluorescence of bacterial cells expressing a variant (GFPm) of the green fluorescent protein (GFP) was reduced to background levels by global replacement of the leucine residues of GFPm by 5,5,5-trifluoroleucine. (
  • It is possible to trace connections between cells, observe reactions between proteins, and to find mechanisms that form signals. (
  • And has boosted the enlargement of exceedingly mechanized live cells fluorescent microscopic systems further which are to be used to monitor cells over the time expressing one or more proteins labelled with fluorescent proteins. (
  • Sorting was gated to recover cells with a fluorescent signal that was diminished compared to the population mode. (
  • We have previously demonstrated that vacuolar sorting receptor (VSR) proteins are concentrated on prevacuolar compartments (PVCs) in plant cells. (
  • PVCs in tobacco ( Nicotiana tabacum ) BY-2 cells are multivesicular bodies (MVBs) as defined by VSR proteins and the BP-80 reporter, where the transmembrane domain (TMD) and cytoplasmic tail (CT) sequences of BP-80 are sufficient and specific for correct targeting of the reporter to PVCs. (
  • Expressing a protein in different bacterial compartments has been shown to affect its accessibility to the immune system and the way the antigen is processed by antigen presenting cells. (
  • To identify the cardiac precursor cells, we have generated stably transfected ES cells with a vector containing the gene of the green fluorescent protein (GFP) under control of the cardiac α-actin promoter. (
  • Such a range of sensitive reporter proteins will greatly improve the prospects for GFP-based applications in cells that require relatively high incubation temperatures. (
  • Split GFPs have been widely applied for imaging as fluorescent reporters of cellular processes, because they are small (∼25 kDa), are stable in cytosol, produce chromophores autocatalytically, and are amenable to mutation and circular permutation ( 4 ). (
  • Previously, it has been shown that the introduction of a second mutation (H148D) into S65T GFP allows the recovery of green emission, implying that ESPT is again possible. (
  • The most important mutation was the replacement of threonine with tyrosine at protein position 203 (T203Y is used to describe the same thing where T and Y represent the single letter code for the amino acids threonine and tyrosine respectively). (
  • Recently, the group of photoactivatable GFP-like proteins attracted particular attention. (
  • Using a unique and relatively simple cell-based fluorescent assay they developed, scientists with the U.S. Department of Energy's Lawrence Berkeley National Laboratory and the University of California, Berkeley have identified a means by which fluoxetine, the active ingredient in Prozac, suppresses the activity of the TREK1 potassium channel. (
  • The green fluorescent protein ( GFP ) is a protein composed of 238 amino acid residues (26.9 kDa ) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. (
  • The success of this approach demonstrates the feasibility of engineering functional proteins containing many copies of abiological amino acid constituents. (
  • GFP is a small protein and has been made up of 238 amino acids. (
  • This jamming event appears to be the result of the conformation assumed by β-galactosidase and not of the presence of specific amino acid sequences that are incompatible with the protein export machinery ( 32 , 47 ). (
  • After an in-frame fusion to the protein of interest, the resulting chimeric protein can be expressed in a cellular environment to monitor function or activity [5] . (
  • The desensitization of GPCR responsiveness results as the culmination of several events including: the uncoupling of receptors from their heterotrimeric G protein as the consequence of receptor phosphorylation, sequestration (endocytosis) of receptors to endosomes and down-regulation of the total cellular complement of receptors. (
  • The advent of highly pH-sensitive fluorescent proteins offers new possibilities for measuring cellular pH ( 11 , 13 , 16 , 22 , 24 ). (
  • Cellular proteins differ widely in their lability, ranging from those that are completely stable to those with half-lives measured in minutes. (
  • Green fluorescent protein has had great success as a marker protein in a variety of biological systems due to its inherent stability, which only adds to its many other desirable characteristics as a marker proteins. (
  • These secretory organelles are defined by specific marker proteins present on organelle membranes. (
  • Ensure accurate, reproducible results with Thermo Scientific GFP (Green Fluorescent Protein) Ab-1, Mouse Monoclonal Antibody. (
  • Hi, I know nothing about green fluorescent protein, but one of our researchers has some retina in formalin that has been labelled with a green fluorescent protein tagged antibody and wants to know what to do with it next. (
  • Purified polyclonal rabbit antibody to Green Fluorescent Protein (GFP) using recombinant GFP expressed in E. coli as immunogen. (
  • Anti green fluorescent protein antibody: 210-PS-1GFP. (
  • Strategies for labelling GBS with fluorescent biomarkers, have so far been limited to antibody-based immunostaining methods and nonspecific protein/DNA stains. (
  • Direct fusion of an antibody to green fluorescent protein (GFP) could overcome these disadvantages. (
  • Conn PM (ed.) (1999) Green Fluorescent Protein Methods in Enzymology, vol. 302. (
  • Later, they study the cell by using special antibodies attached to the green fluorescent proteins. (
  • As an example, immunoprecipitation experiments using antibodies directed against ActA have indicated that the protein becomes one of the most abundant bacterial surface proteins expressed by intracytoplasmic L. monocytogenes ( 7 ). (
  • Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum . (
  • Using a submarine-based and -operated fluorescence detection system in the Gulf of Mexico, we discovered ceriantharians emitting bright green fluorescence in depths between 500 and 600 m and identified a GFP, named cerFP505, with bright fluorescence emission peaking at 505 nm. (
  • Desensitization in response to GRK-mediated phosphorylation involves the binding of arrestin proteins that serve to sterically uncouple the receptor from its G protein. (
  • Toward this goal, we have generated seven chimeric constructs containing signal peptide (sp) linked to green fluorescent protein (GFP) and TMD/CT sequences (sp-GFP-TMD/CT) of the seven individual AtVSR. (
  • and low fluorescent, LF), characterized by markedly different contents of a cyan fluorescent protein, were exposed to different quantities of blue light (470 nm) that matched the major absorption band of the host pigment (473 nm). (
  • While the wild-type GFP protein has not been particularly useful as a sensor in FRET, several mutants of GFP have been manufactured to create proteins with distinct fluorescence spectra. (
  • Despite the successes of using β-galactosidase fusion proteins for genetic analysis of protein export, biases and limitations to the genetic selections and screens that use this reporter likely exist, potentially limiting the spectrum of export mutants that can be isolated. (
  • Scientists Roger Y. Tsien , Osamu Shimomura , and Martin Chalfie were awarded the 2008 Nobel Prize in Chemistry on 10 October 2008 for their discovery and development of the green fluorescent protein. (
  • In order to obtain the required protein in a soluble and functional format, IBs usually undergo solubilization and refolding processes, which are expensive, time consuming and highly proteindependent [2]. (
  • Availability of a functional β 2 AR tagged with the highly sensitive Green Fluorescent Protein (GFP) could allow measurements of the various properties of the β 2 AR. (
  • Fluorescent proteins provide highly sensitive detection, do not require indicator loading, and lack phototoxicity ( 5 , 11 , 20 , 23 ). (
  • We report a highly effective on-chip preconcentration method by combining field-amplified sample injection (FASI) and bovine serum albumin (BSA) sweeping for ultrasensitive detection of green fluorescent protein (GFP) on a simple cross-channel microchip device. (
  • However, the concentration limits of detection of μ CE are often inadequate for the measurement of proteins/peptides at low concentration levels. (
  • 14 - 17 For protein analysis, field-amplified stacking can improve the limit of detection considerably, e.g. by 30-fold for albumin. (
  • These reporters have been especially valuable in identifying regions of a protein that promote membrane translocation ( 6 , 24 ) and allow predictions of membrane topology ( 37 , 38 ). (
  • G protein-coupled receptors (GPCRs) form the largest family of integral membrane receptors. (
  • This study may serve as a general guide for improving the yields of other membrane protein preparations and selecting the appropriate detergent to stabilize membrane proteins for biophysical and biochemical analyses. (
  • Differential fluorescence/glycosylation pattern probes membrane protein topology. (