Temperate bacteriophage of the genus INOVIRUS which infects enterobacteria, especially E. coli. It is a filamentous phage consisting of single-stranded DNA and is circularly permuted.
Viruses whose hosts are bacterial cells.
Viruses whose host is Escherichia coli.
Virulent bacteriophage and type species of the genus T4-like phages, in the family MYOVIRIDAE. It infects E. coli and is the best known of the T-even phages. Its virion contains linear double-stranded DNA, terminally redundant and circularly permuted.
Deoxyribonucleic acid that makes up the genetic material of viruses.
Viruses whose nucleic acid is DNA.
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
A temperate inducible phage and type species of the genus lambda-like viruses, in the family SIPHOVIRIDAE. Its natural host is E. coli K12. Its VIRION contains linear double-stranded DNA with single-stranded 12-base 5' sticky ends. The DNA circularizes on infection.
Proteins found in any species of virus.
Virulent bacteriophage and type species of the genus T7-like phages, in the family PODOVIRIDAE, that infects E. coli. It consists of linear double-stranded DNA, terminally redundant, and non-permuted.
The phenomenon by which a temperate phage incorporates itself into the DNA of a bacterial host, establishing a kind of symbiotic relation between PROPHAGE and bacterium which results in the perpetuation of the prophage in all the descendants of the bacterium. Upon induction (VIRUS ACTIVATION) by various agents, such as ultraviolet radiation, the phage is released, which then becomes virulent and lyses the bacterium.
A single chain of deoxyribonucleotides that occurs in some bacteria and viruses. It usually exists as a covalently closed circle.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
The functional hereditary units of VIRUSES.
A series of 7 virulent phages which infect E. coli. The T-even phages T2, T4; (BACTERIOPHAGE T4), and T6, and the phage T5 are called "autonomously virulent" because they cause cessation of all bacterial metabolism on infection. Phages T1, T3; (BACTERIOPHAGE T3), and T7; (BACTERIOPHAGE T7) are called "dependent virulent" because they depend on continued bacterial metabolism during the lytic cycle. The T-even phages contain 5-hydroxymethylcytosine in place of ordinary cytosine in their DNA.
A temperate coliphage, in the genus Mu-like viruses, family MYOVIRIDAE, composed of a linear, double-stranded molecule of DNA, which is able to insert itself randomly at any point on the host chromosome. It frequently causes a mutation by interrupting the continuity of the bacterial OPERON at the site of insertion.
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.
The process by which a DNA molecule is duplicated.
Virulent bacteriophage and sole member of the genus Cystovirus that infects Pseudomonas species. The virion has a segmented genome consisting of three pieces of doubled-stranded DNA and also a unique lipid-containing envelope.
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.
Any of the covalently closed DNA molecules found in bacteria, many viruses, mitochondria, plastids, and plasmids. Small, polydisperse circular DNA's have also been observed in a number of eukaryotic organisms and are suggested to have homology with chromosomal DNA and the capacity to be inserted into, and excised from, chromosomal DNA. It is a fragment of DNA formed by a process of looping out and deletion, containing a constant region of the mu heavy chain and the 3'-part of the mu switch region. Circular DNA is a normal product of rearrangement among gene segments encoding the variable regions of immunoglobulin light and heavy chains, as well as the T-cell receptor. (Riger et al., Glossary of Genetics, 5th ed & Segen, Dictionary of Modern Medicine, 1992)
Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.
The type species of the genus MICROVIRUS. A prototype of the small virulent DNA coliphages, it is composed of a single strand of supercoiled circular DNA, which on infection, is converted to a double-stranded replicative form by a host enzyme.
The outer protein protective shell of a virus, which protects the viral nucleic acid.
Stable phosphorus atoms that have the same atomic number as the element phosphorus, but differ in atomic weight. P-31 is a stable phosphorus isotope.
A species of temperate bacteriophage in the genus P2-like viruses, family MYOVIRIDAE, which infects E. coli. It consists of linear double-stranded DNA with 19-base sticky ends.
Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.
A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.
Proteins that form the CAPSID of VIRUSES.
Bacteriophage in the genus T7-like phages, of the family PODOVIRIDAE, which is very closely related to BACTERIOPHAGE T7.
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 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.
A technique of bacterial typing which differentiates between bacteria or strains of bacteria by their susceptibility to one or more bacteriophages.
A species of temperate bacteriophage in the genus P1-like viruses, family MYOVIRIDAE, which infects E. coli. It is the largest of the COLIPHAGES and consists of double-stranded DNA, terminally redundant, and circularly permuted.
Viruses whose host is Salmonella. A frequently encountered Salmonella phage is BACTERIOPHAGE P22.
A family of BACTERIOPHAGES and ARCHAEAL VIRUSES which are characterized by long, non-contractile tails.
The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.
A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.
Bacteriophages whose genetic material is RNA, which is single-stranded in all except the Pseudomonas phage phi 6 (BACTERIOPHAGE PHI 6). All RNA phages infect their host bacteria via the host's surface pili. Some frequently encountered RNA phages are: BF23, F2, R17, fr, PhiCb5, PhiCb12r, PhiCb8r, PhiCb23r, 7s, PP7, Q beta phage, MS2 phage, and BACTERIOPHAGE PHI 6.
Spectroscopic method of measuring the magnetic moment of elementary particles such as atomic nuclei, protons or electrons. It is employed in clinical applications such as NMR Tomography (MAGNETIC RESONANCE IMAGING).
Rupture of bacterial cells due to mechanical force, chemical action, or the lytic growth of BACTERIOPHAGES.
Bacteriophage and type species in the genus Tectivirus, family TECTIVIRIDAE. They are specific for Gram-negative bacteria.
Viruses whose host is Pseudomonas. A frequently encountered Pseudomonas phage is BACTERIOPHAGE PHI 6.
Viruses whose host is Staphylococcus.
Viruses whose host is Bacillus. Frequently encountered Bacillus phages include bacteriophage phi 29 and bacteriophage phi 105.
Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)
A family of bacteriophages which are characterized by short, non-contractile tails.
Viruses whose host is Streptococcus.
Proteins found in the tail sections of DNA and RNA viruses. It is believed that these proteins play a role in directing chain folding and assembly of polypeptide chains.
A bacteriophage genus of the family LEVIVIRIDAE, whose viruses contain the short version of the genome and have a separate gene for cell lysis.
The complete genetic complement contained in a DNA or RNA molecule in a virus.
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).
The adhesion of gases, liquids, or dissolved solids onto a surface. It includes adsorptive phenomena of bacteria and viruses onto surfaces as well. ABSORPTION into the substance may follow but not necessarily.
The folding of an organism's DNA molecule into a compact, orderly structure that fits within the limited space of a CELL or VIRUS PARTICLE.
Genomes of temperate BACTERIOPHAGES integrated into the DNA of their bacterial host cell. The prophages can be duplicated for many cell generations until some stimulus induces its activation and virulence.
A genus of filamentous bacteriophages of the family INOVIRIDAE. Organisms of this genus infect enterobacteria, PSEUDOMONAS; VIBRIO; and XANTHOMONAS.
A subdiscipline of genetics which deals with the genetic mechanisms and processes of microorganisms.
Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992).
Specific loci on both the bacterial DNA (attB) and the phage DNA (attP) which delineate the sites where recombination takes place between them, as the phage DNA becomes integrated (inserted) into the BACTERIAL DNA during LYSOGENY.
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.
Method for measuring viral infectivity and multiplication in CULTURED CELLS. Clear lysed areas or plaques develop as the VIRAL PARTICLES are released from the infected cells during incubation. With some VIRUSES, the cells are killed by a cytopathic effect; with others, the infected cells are not killed but can be detected by their hemadsorptive ability. Sometimes the plaque cells contain VIRAL ANTIGENS which can be measured by IMMUNOFLUORESCENCE.
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.
A species of gram-positive bacteria that is a common soil and water saprophyte.
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.
Deoxyribonucleic acid that makes up the genetic material of bacteria.
Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.
A family of bacteriophages containing one genus (Cystovirus) with one member (BACTERIOPHAGE PHI 6).
A species of filamentous Pseudomonas phage in the genus INOVIRUS, family INOVIRIDAE.
Enzymes which catalyze the hydrolases of ester bonds within DNA. EC 3.1.-.
An antibiotic first isolated from cultures of Streptomyces venequelae in 1947 but now produced synthetically. It has a relatively simple structure and was the first broad-spectrum antibiotic to be discovered. It acts by interfering with bacterial protein synthesis and is mainly bacteriostatic. (From Martindale, The Extra Pharmacopoeia, 29th ed, p106)
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.
Any method used for determining the location of and relative distances between genes on a chromosome.
An order comprising three families of tailed bacteriophages: MYOVIRIDAE; PODOVIRIDAE; and SIPHOVIRIDAE.
Ribonucleic acid that makes up the genetic material of viruses.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
Proteins found in any species of bacterium.
DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair.
A test used to determine whether or not complementation (compensation in the form of dominance) will occur in a cell with a given mutant phenotype when another mutant genome, encoding the same mutant phenotype, is introduced into that cell.
A single-stranded DNA-dependent RNA polymerase that functions to initiate, or prime, DNA synthesis by synthesizing oligoribonucleotide primers. EC 2.7.7.-.
Treatment of diseases with biological materials or biological response modifiers, such as the use of GENES; CELLS; TISSUES; organs; SERUM; VACCINES; and humoral agents.
Electron microscopy involving rapid freezing of the samples. The imaging of frozen-hydrated molecules and organelles permits the best possible resolution closest to the living state, free of chemical fixatives or stains.
The properties of a pathogen that makes it capable of infecting one or more specific hosts. The pathogen can include PARASITES as well as VIRUSES; BACTERIA; FUNGI; or PLANTS.
Enzymes that catalyze the incorporation of deoxyribonucleotides into a chain of DNA. EC 2.7.7.-.
Refuse liquid or waste matter carried off by sewers.
The sum of the weight of all the atoms in a molecule.
The functional hereditary units of BACTERIA.
Macromolecular molds for the synthesis of complementary macromolecules, as in DNA REPLICATION; GENETIC TRANSCRIPTION of DNA to RNA, and GENETIC TRANSLATION of RNA into POLYPEPTIDES.
A broad category of viral proteins that play indirect roles in the biological processes and activities of viruses. Included here are proteins that either regulate the expression of viral genes or are involved in modifying host cell functions. Many of the proteins in this category serve multiple functions.
The rate dynamics in chemical or physical systems.
Viral proteins that are components of the mature assembled VIRUS PARTICLES. They may include nucleocapsid core proteins (gag proteins), enzymes packaged within the virus particle (pol proteins), and membrane components (env proteins). These do not include the proteins encoded in the VIRAL GENOME that are produced in infected cells but which are not packaged in the mature virus particle,i.e. the so called non-structural proteins (VIRAL NONSTRUCTURAL PROTEINS).
In bacteria, a group of metabolically related genes, with a common promoter, whose transcription into a single polycistronic MESSENGER RNA is under the control of an OPERATOR REGION.
Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.
Tungsten hydroxide oxide phosphate. A white or slightly yellowish-green, slightly efflorescent crystal or crystalline powder. It is used as a reagent for alkaloids and many other nitrogen bases, for phenols, albumin, peptone, amino acids, uric acid, urea, blood, and carbohydrates. (From Merck Index, 11th ed)
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
Disruption of the secondary structure of nucleic acids by heat, extreme pH or chemical treatment. Double strand DNA is "melted" by dissociation of the non-covalent hydrogen bonds and hydrophobic interactions. Denatured DNA appears to be a single-stranded flexible structure. The effects of denaturation on RNA are similar though less pronounced and largely reversible.
A sequence of successive nucleotide triplets that are read as CODONS specifying AMINO ACIDS and begin with an INITIATOR CODON and end with a stop codon (CODON, TERMINATOR).
That portion of the electromagnetic spectrum immediately below the visible range and extending into the x-ray frequencies. The longer wavelengths (near-UV or biotic or vital rays) are necessary for the endogenous synthesis of vitamin D and are also called antirachitic rays; the shorter, ionizing wavelengths (far-UV or abiotic or extravital rays) are viricidal, bactericidal, mutagenic, and carcinogenic and are used as disinfectants.
Biologically active DNA which has been formed by the in vitro joining of segments of DNA from different sources. It includes the recombination joint or edge of a heteroduplex region where two recombining DNA molecules are connected.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
A group of methylazirinopyrroloindolediones obtained from certain Streptomyces strains. They are very toxic antibiotics used as ANTINEOPLASTIC AGENTS in some solid tumors. PORFIROMYCIN and MITOMYCIN are the most useful members of the group.
Catalyze the joining of preformed ribonucleotides or deoxyribonucleotides in phosphodiester linkage during genetic processes. EC 6.5.1.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Viruses whose host is one or more Mycobacterium species. They include both temperate and virulent types.
A group of enzymes catalyzing the endonucleolytic cleavage of DNA. They include members of EC 3.1.21.-, EC 3.1.22.-, EC 3.1.23.- (DNA RESTRICTION ENZYMES), EC 3.1.24.- (DNA RESTRICTION ENZYMES), and EC 3.1.25.-.
The effects of ionizing and nonionizing radiation upon living organisms, organs and tissues, and their constituents, and upon physiologic processes. It includes the effect of irradiation on food, drugs, and chemicals.
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.
The regulatory elements of an OPERON to which activators or repressors bind thereby effecting the transcription of GENES in the operon.
The assembly of VIRAL STRUCTURAL PROTEINS and nucleic acid (VIRAL DNA or VIRAL RNA) to form a VIRUS PARTICLE.
Enzymes that catalyze the release of mononucleotides by the hydrolysis of the terminal bond of deoxyribonucleotide or ribonucleotide chains.
Proteins that catalyze the unwinding of duplex DNA during replication by binding cooperatively to single-stranded regions of DNA or to short regions of duplex DNA that are undergoing transient opening. In addition DNA helicases are DNA-dependent ATPases that harness the free energy of ATP hydrolysis to translocate DNA strands.
A non-pathogenic species of LACTOCOCCUS found in DAIRY PRODUCTS and responsible for the souring of MILK and the production of LACTIC ACID.
A large family of lytic bacteriophages infecting enterobacteria; SPIROPLASMA; BDELLOVIBRIO; and CHLAMYDIA. It contains four genera: MICROVIRUS; Spiromicrovirus; Bdellomicrovirus; and Chlamydiamicrovirus.
Enzymes that catalyze the template-directed incorporation of ribonucleotides into an RNA chain. EC 2.7.7.-.
A serotype of Salmonella enterica that is a frequent agent of Salmonella gastroenteritis in humans. It also causes PARATYPHOID FEVER.
Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.
The infective system of a virus, composed of the viral genome, a protein core, and a protein coat called a capsid, which may be naked or enclosed in a lipoprotein envelope called the peplos.
Proteins obtained from ESCHERICHIA COLI.

Identification of residues in beta-lactamase critical for binding beta-lactamase inhibitory protein. (1/356)

beta-Lactamase inhibitory protein (BLIP) is a potent inhibitor of several beta-lactamases including TEM-1 beta-lactamase (Ki = 0.1 nM). The co-crystal structure of TEM-1 beta-lactamase and BLIP has been solved, revealing the contact residues involved in the interface between the enzyme and inhibitor. To determine which residues in TEM-1 beta-lactamase are critical for binding BLIP, the method of monovalent phage display was employed. Random mutants of TEM-1 beta-lactamase in the 99-114 loop-helix and 235-240 B3 beta-strand regions were displayed as fusion proteins on the surface of the M13 bacteriophage. Functional mutants were selected based on the ability to bind BLIP. After three rounds of enrichment, the sequences of a collection of functional beta-lactamase mutants revealed a consensus sequence for the binding of BLIP. Seven loop-helix residues including Asp-101, Leu-102, Val-103, Ser-106, Pro-107, Thr-109, and His-112 and three B3 beta-strand residues including Ser-235, Gly-236, and Gly-238 were found to be critical for tight binding of BLIP. In addition, the selected beta-lactamase mutants A113L/T114R and E240K were found to increase binding of BLIP by over 6- and 11-fold, respectively. Combining these substitutions resulted in 550-fold tighter binding between the enzyme and BLIP with a Ki of 0.40 pM. These results reveal that the binding between TEM-1 beta-lactamase and BLIP can be improved and that there are a large number of sequences consistent with tight binding between BLIP and beta-lactamase.  (+info)

Gene transfer to mammalian cells using genetically targeted filamentous bacteriophage. (2/356)

We have genetically modified filamentous bacteriophage to deliver genes to mammalian cells. In previous studies we showed that noncovalently attached fibroblast growth factor (FGF2) can target bacteriophage to COS-1 cells, resulting in receptor-mediated transduction with a reporter gene. Thus, bacteriophage, which normally lack tropism for mammalian cells, can be adapted for mammalian cell gene transfer. To determine the potential of using phage-mediated gene transfer as a novel display phage screening strategy, we transfected COS-1 cells with phage that were engineered to display FGF2 on their surface coat as a fusion to the minor coat protein, pIII. Immunoblot and ELISA analysis confirmed the presence of FGF2 on the phage coat. Significant transduction was obtained in COS-1 cells with the targeted FGF2-phage compared with the nontargeted parent phage. Specificity was demonstrated by successful inhibition of transduction in the presence of excess free FGF2. Having demonstrated mammalian cell transduction by phage displaying a known gene targeting ligand, it is now feasible to apply phage-mediated transduction as a screen for discovering novel ligands.  (+info)

Selection-dominant and nonaccessible epitopes on cell-surface receptors revealed by cell-panning with a large phage antibody library. (3/356)

To generate antibodies to defined cell-surface antigens, we used a large phage antibody fragment library to select on cell transfectants expressing one of three chosen receptors. First, in vitro panning procedures and phage antibody screening ELISAs were developed using whole live cells stably expressing the antigen of interest. When these methodologies were applied to Chinese hamster ovary (CHO) cells expressing one of the receptors for a neuropeptide, somatostatin, using either direct cell panning or a strategy of depletion or ligand-directed elution, many different pan-CHO-cell binders were selected, but none was receptor specific. However, when using direct panning on CHO-cells expressing the human membrane protein CD36, an extraordinary high frequency of antigen-specific phage antibodies was found. Panning on myoblasts expressing the rat homologue of CD36 revealed a similar selection dominance for anti-(CD36). Binding of all selected 20 different anti-(CD36) phage was surprisingly inhibited by one anti-(CD36) mAb CLB-IVC7, which recognizes a functional epitope that is also immunodominant in vivo. Similar inhibition was found for seven anti-(rat) CD36 that cross-reacted with human CD36. Our results show that, although cells can be used as antigen carriers to select and screen phage antibodies, the nature of the antigen target has a profound effect on the outcome of the selection.  (+info)

Identification of peroxisomal proteins by using M13 phage protein VI phage display: molecular evidence that mammalian peroxisomes contain a 2,4-dienoyl-CoA reductase. (4/356)

To elucidate unknown mammalian peroxisomal enzymes and functions, we subjected M13 phage expressing fusions between the gene encoding protein VI and a rat liver cDNA library to an immunoaffinity selection process in vitro (biopanning) with the use of antibodies raised against peroxisomal subfractions. In an initial series of biopanning experiments, four different cDNA clones were obtained. These cDNA species encoded two previously identified peroxisomal enzymes, catalase and urate oxidase, and two novel proteins that contained a C-terminal peroxisomal targeting signal (PTS1). A primary structure analysis of these novel proteins revealed that one, ending in the tripeptide AKL, is homologous to the yeast peroxisomal 2,4-dienoyl-CoA reductase (EC 1.3.1.34; DCR), an enzyme required for the degradation of unsaturated fatty acids, and that the other, ending in the tripeptide SRL, is a putative member of the short-chain dehydrogenase/reductase (SDR) family, with three isoforms. Green fluorescent protein (GFP) fusions encoding GFP-DCR-AKL, GFP-DCR, GFP-SDR-SRL and GFP-SDR were expressed in mammalian cells. The analysis of the subcellular location of the recombinant fusion proteins confirmed the peroxisomal localization of GFP-DCR-AKL and GFP-SDR-SRL, as well as the functionality of the PTS1. That the AKL protein is indeed an NADPH-dependent DCR was demonstrated by showing DCR activity of the bacterially expressed protein. These results demonstrate at the molecular level that mammalian peroxisomes do indeed contain a DCR. In addition, the results presented here indicate that the protein VI display system is suitable for the isolation of rare cDNA clones from cDNA libraries and that this technology facilitates the identification of novel peroxisomal proteins.  (+info)

Peptide ligands to human immunodeficiency virus type 1 gp120 identified from phage display libraries. (5/356)

We have used phage-displayed peptide libraries to identify novel ligands to the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein gp120. Screening of libraries of random 12-mers, 7-mers, and cyclic 9-mers produced two families of gp120 binding peptides. Members of a family with the prototype sequence RINNIPWSEAMM (peptide 12p1) inhibit the interaction between gp120 and both four-domain soluble CD4 (4dCD4) and monoclonal antibody (MAb) 17b, a neutralizing antibody that covers the chemokine receptor binding surface on gp120. Peptide 12p1 inhibits the interaction of 4dCD4 with gp120 from three different HIV strains, implying that it binds to a conserved site on gp120. Members of a second family of peptides, with the prototype sequence TSPYEDWQTYLM (peptide 12p2), bind more weakly to gp120. They do not detectably affect its interaction with 4dCD4, but they enhance its binding to MAb 17b. A common sequence motif in the two peptide families and cross-competition for gp120 binding suggest that they have overlapping contacts. Their divergent effects on the affinity of gp120 for MAb 17b may indicate that their binding stabilizes distinct conformational states of gp120. The functional properties of 12p1 suggest that it might be a useful lead for the development of inhibitors of HIV entry.  (+info)

Peptide ligands that bind IgM antibodies and block interaction with antigen. (6/356)

We have selected a peptide-display phage library on IgM Abs and identified a panel of phage-expressing peptides that bind to IgM Abs in general, but not to Abs of other classes. A synthetic peptide corresponding to one of the displayed peptide sequences also binds to IgM Abs. The peptides bind to both soluble pentameric Abs and to monomeric cell-surface IgM. The phage-displayed and synthetic peptides inhibit the binding of IgM Abs to Ag. These peptides may create confounding artifacts when IgM Abs are used for epitope mapping studies. Nonetheless, the peptides may have both experimental and therapeutic utility.  (+info)

Selection of a C5a receptor antagonist from phage libraries attenuating the inflammatory response in immune complex disease and ischemia/reperfusion injury. (7/356)

A C5a-receptor antagonist was selected from human C5a phage display libraries in which the C terminus of des-Arg74-hC5a was mutated. The selected molecule is a competitive C5a receptor antagonist in vitro and in vivo. Signal transduction is interrupted at the level of G-protein activation. In addition, the antagonist does not cause any C5a receptor phosphorylation. Proinflammatory properties such as chemotaxis or lysosomal enzyme release of differentiated U937 cells, as well as C5a-induced changes in intracellular Ca2+ concentration of murine peritoneal macrophages, are inhibited. The in vivo efficacy was evaluated in three different animal models of immune complex diseases in mice, i.e., the reverse passive Arthus reaction in the peritoneum, skin, and lung. The i.v. application of the C5a receptor antagonist abrogated polymorphonuclear neutrophil accumulation in peritoneum and markedly attenuated polymorphonuclear neutrophil migration into the skin and the lung. In a model of intestinal ischemia/reperfusion injury, i.v. administration of the C5a receptor antagonist decreased local and remote tissue injury: bowel wall edema and hemorrhage as well as pulmonary microvascular dysfunction. These data give evidence that C5a is an important mediator triggering the inflammatory sequelae seen in immune complex diseases and ischemia/reperfusion injury. The selected C5a receptor antagonist may prove useful to attenuate the inflammatory response in these disorders.  (+info)

New inhibitors of Helicobacter pylori urease holoenzyme selected from phage-displayed peptide libraries. (8/356)

Urease is an important virulence factor for Helicobacter pylori and is critical for bacterial colonization of the human gastric mucosa. Specific inhibition of urease activity has been proposed as a possible strategy to fight this bacteria which infects billions of individual throughout the world and can lead to severe pathological conditions in a limited number of cases. We have selected peptides which specifically bind and inhibit H. pylori urease from libraries of random peptides displayed on filamentous phage in the context of pIII coat protein. Screening of a highly diverse 25-mer combinatorial library and two newly constructed random 6-mer peptide libraries on solid phase H. pylori urease holoenzyme allowed the identification of two peptides, 24-mer TFLPQPRCSALLRYLSEDGVIVPS and 6-mer YDFYWW that can bind and inhibit the activity of urease purified from H. pylori. These two peptides were chemically synthesized and their inhibition constants (Ki) were found to be 47 microM for the 24-mer and 30 microM for the 6-mer peptide. Both peptides specifically inhibited the activity of H. pylori urease but not that of Bacillus pasteurii.  (+info)

The major coat protein of bacteriophage M13 is synthesized as a precursor, the procoat, with a typical leader (signal) sequence of 23 residues at its NH2-terminus. A fusion protein that contains the NH2-terminal 141 residues of cytoplasmic ribulokinase and all but the first ten residues of M13 procoat was made. The fusion protein inserts into the plasma membrane of Escherichia coli and is processed by leader peptidase to give rise to a leader peptide of 155 residues and the mature coat protein of 50 residues. The NH2-terminus of the leader peptide remains in the cytoplasm and is protected from protease added to the medium outside of the cell. This indicates that M13 procoat inserts into the membrane as a loop structure and that the NH2-terminus of a leader peptide remains within the cytoplasm during membrane insertion. ...
With a mass of ~1.6 x 107 Daltons and com- posed of approximately 2700 proteins, bacteriophage M13 has been employed as a molecular scaffold in bionanomaterials fabrication. In order to extend the versatility of M13 in this area, residue-specific unnatural amino acid incorporation was employed to successfully display azide functionalities on specific solvent-exposed positions of the pVIII major sheath protein of this bacteriophage. Employing a combination of engineered mutants of the gene coding for the pVIII protein, the methionine (Met) analog, L-azidohomoalanine (Aha), and a suitable Escherichia coli Met auxotroph for phage pro- duction, conditions were developed to produce M13 bacteri- ophage labeled with over 350 active azides (estimated by fluorescent dye labeling utilizing a strain-promoted azide- alkyne cycloaddition) and capable of azide-selective attach- ment to 5 nm gold nanoparticles as visualized by transmis- sion electron microscopy. The capability of this system to undergo dual ...
Mouse anti-M13 phage coat protein g8p. Antibodies recognising M13 filamentous phage coat proteins are instrumental in the selection and detection of phages expressing specific antibody fragments or peptide sequences at their surface. The monoclonal antibodies manufactured and supplied by Exalpha react with either the pIII (g3p) or pVIII (g8p) proteins of M13 filamentous bacteriophage. All antibodies are available in a purified format. The antibodies are fully validated and are suitable for a wide range of techniques including ...
Mouse anti-M13 phage coat protein g8p. Antibodies recognising M13 filamentous phage coat proteins are instrumental in the selection and detection of phages expressing specific antibody fragments or peptide sequences at their surface. The monoclonal antibodies manufactured and supplied by Exalpha react with either the pIII (g3p) or pVIII (g8p) proteins of M13 filamentous bacteriophage. All antibodies are available in a purified format. The antibodies are fully validated and are suitable for a wide range of techniques including ...
The goal of therapy was never integration. I was happy living as a fragmented person. Me and Richard accidently triggered spontaneous integration and once that door was opened the smaller parts fused to others and then as time went on those no longer needed fused to the core too. There are only a handful that I still feel rolling around with in me. Remnants of a ancient life that seems very distant now. Levels of my consciousness that enrich me as a whole. Dissociation/fracturing out a skill I can still call upon even today ...
Having been researching in the field of phage display technology for years, Creative Biolabs is able to offer a series of comprehensive phage display library screening services by its own biopanning strategies and enhanced equipments.. Phage display technology is one of the major approaches applied in various protein interaction studies, especially in the discovery of specific antibodies, scaffolds and ligands. As binders with the desired specificity generally exist at low frequencies in the constructed libraries, phage display library screening has become an effective technique for the enrichment and identification of binders with high affinity and specificity from interested libraries.. Libraries from customized library construction services, in-house premade antibody libraries, peptide libraries or scaffold libraries, libraries provided by clients and other commercialized libraries can all be screened in Creative Biolabs.. No matter its purified or unpurified targets, we can screen for ...
In vivo screening of phage-displayed peptide libraries has revealed extensive molecular differences in the blood vessels of individual normal tissues. Pathological lesions also put their signature on the vasculature; in tumours, both blood and lymphatic vessels differ from normal vessels. The changes that characterize tumour blood vessels include selective expression of certain integrins. Peptides isolated by in vivo phage display for homing to tumours have been shown to be useful in directing therapeutic agents to experimental tumours. The targeting can enhance the efficacy of the therapy while reducing side effects. Phage screening has also revealed lung-specific vascular markers that promote tumour metastasis to the lungs by mediating specific adherence of tumour cells to the lung vasculature. These phage-screening studies have revealed a previously unsuspected degree of vascular specialization and provide potentially useful guidance devices for targeted therapies.. ...
Since the advent of phage display technology, dating back to 1985, antibody libraries displayed on filamentous phage surfaces have been used to identify specific binders for many different purposes, including the recognition of tumors. Phage display represents a high-throughput technique for screening billions of random fusion antibodies against virtually any target on the surface or inside cancer cells, or even soluble markers found in patient serum. Many phage display derived binders targeting important tumor markers have been identified. Selection directed to tumoral cells surfaces lead to the identification of unknown tumoral markers. Also the improvement of methods that require smaller amounts of cells has opened the possibility to use this approach on patient samples. Robust techniques combining an antibody library displayed on the phage surface and protein microarray allowed the identification of auto antibodies recognized by patient sera. Many Ab molecules directly or indirectly targeting
628 Nasopharyngeal carcinoma (NPC) is one of the most common cancers among Chinese living in South China, Taiwan and Singapore. For treatment of NPC, although radiotherapy, surgical removal, and chemotherapy have been applied for decades and the 5 year survival rate in the advanced cases are still below 75 %. Much effort using other methods such as high dose chemotherapy plus bone marrow stem cell injection and other targeting therapy are needed to control this cancer. In the present experiment, we used random peptide phage display libraries to identify and isolate a specific novel peptide from NPC cell line for targeting drug delivery. After subtraction with control epithelial cells for 3 times, the remained phage libraries were biopanning on NPC-TW04 cells for 5 rounds. It was found that 44 selected phage clones could specifically bind to NPC cell surface. When ELISA was used to verify those clones, 16 phage clones showed higher affinity to NPC cells and were subjected to DNA sequencing. The ...
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A phage display library was used to identify peptide sequences that target bone. The objective was to investigate the effect of these...
The time frame for taking action and reporting SSBA handling to the department is within two business days (the exception to this is the Regular Report). This time frame means that if you receive an SSBA or suspected SSBA on Monday you must take action (e.g. register to handle, transfer or destroy the SSBA) and report the action taken no later than close of business on Wednesday (Monday is considered the date of receipt and is therefore excluded in the two-business-day calculation). Similarly, if you receive an SSBA or suspected SSBA on Saturday you are not required to report or take action until close of business Wednesday (as the SSBA was received on Saturday, Monday is considered the date of receipt and is therefore excluded in the two-businessday calculation). During this time you must comply with the relevant parts of the SSBA Standards ...
The first step to producing our specially designed enzymes was to change the wild-type gene that codes for Kumamolisin to code instead for variant enzymes with our desired amino acid substitutions. We designed mutagenic oligonucleotide primers that would anneal to the wild-type Kumamolisin gene and incorporate point mutations that, when expressed, would result in a variant of Kumamolisin with the desired amino acid shift. To incorporate these mutations, we first isolated single stranded DNA (ssDNA) of our vector harboring the wild-type Kumamolisin gene. To do this we infected cells with bacteriophage M13, which packages its own ssDNA genome identified by length, and so in tandem packaged our vector in single stranded form. We then harvested the phage from the lysed culture of E. coli, and extracted our single stranded vector DNA. Next, we annealed and extended our mutagenic oligos to incorporate the specified mutations into the newly synthesized antisense strand. This hybrid vector was ...
The first step to producing our specially designed enzymes was to change the wild-type gene that codes for Kumamolisin to code instead for variant enzymes with our desired amino acid substitutions. We designed mutagenic oligonucleotide primers that would anneal to the wild-type Kumamolisin gene and incorporate point mutations that, when expressed, would result in a variant of Kumamolisin with the desired amino acid shift. To incorporate these mutations, we first isolated single stranded DNA (ssDNA) of our vector harboring the wild-type Kumamolisin gene. To do this we infected cells with bacteriophage M13, which packages its own ssDNA genome identified by length, and so in tandem packaged our vector in single stranded form. We then harvested the phage from the lysed culture of E. coli, and extracted our single stranded vector DNA. Next, we annealed and extended our mutagenic oligos to incorporate the specified mutations into the newly synthesized antisense strand. This hybrid vector was ...
Hi, I wonder if anybody can give me a clue to make ssDNA of pET-based plasmid with good yield. We have tried Novablue and XL1-blue as host cells and USC-M13 as a helper phage without success. I was told to use Promegas R408 helper phage instead. Any input ? Thanks in advance. Yee-yung Charng yccharng at ucdavis.edu ...
Bacteriophage M13 major coat protein has been incorporated at different lipid/protein ratios in lipid bilayers consisting of various ratios of dimyristoylphosphatidylcholine (DMPC) to dimyristoylphosphatidylglycerol (DMPG). Spin-label ESR experiments were performed with phospholipids labeled at the C-14 position of the sn-2 chain. For M13 coat protein recombinants with DMPC alone, the relative association constants were determined for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels and found to be 1.0, 1.0, and 2.1 relative to the background DMPC, respectively. The number of association sites for each phospholipid on the protein was found to be 4 per protein monomer. The intrinsic off-rates for lipid exchange at the intramembranous surface of the protein in DMPC alone at 30 degrees C were found to be 5 X 10(6), 6 X 10(6), and 2 X 10(6) s-1 for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels, respectively. Adding DMPG to the DMPC lipid
Dr. Marilena Hall received her B.S. in Chemistry in 1992 from McGill University in Montreal, Canada. She then earned her Ph.D. at the California Institute of Technology (NSERC Fellowship) where she worked in the laboratory of Dr. Jacqueline K. Barton. Her thesis involved the design of a synthetic deoxyribonuclease using a chimera of a DNA-binding rhodium complex and a short Zn2+ -coordinating peptide.. From 1998-2000, Dr. Hall carried out post-doctoral research at New England Biolabs in Beverly, MA creating an artificial bifunctional intein capable of both protein splicing and homing endonuclease activity. During her post-doc, she was also an adjunct professor at Massasoit Community College, teaching general chemistry from 1999-2000. Professor Hall joined the Stonehill Department of Chemistry in the Fall of 2000.. Professor Halls research program employs peptide phage display libraries to identify short peptides that can mimic the coordination of Zn2+ in zinc-containing metalloenzymes.. Several ...
article{c02feb63-0c75-498d-9b63-6043d6df0ab2, abstract = {Background: Phage Display technology is a well established technique for high throughput screening of affinity ligands. Here we describe a new compact chromato-panning procedure for selection of suitable binders from a phage peptide display library. Results: Both phages and E. coli cells pass non-hindered through the interconnected pores of macroporous gel, so called cryogel. After coupling a ligand to a monolithic cryogel column, the phage library was applied on the column and non-bound phages were washed out. The selection of strong phage-binders was achieved already after the first panning cycle due to the efficient separation of phage-binders from phage-non-binders in chromatographic mode rather than in batch mode as in traditional biopanning procedures. E. coli cells were applied on the column for infection with the specifically bound phages. Conclusion: Chromato-panning allows combining several steps of the panning procedure ...
Synthesis of Single-stranded DNA Probes of Heterogeneous Length from Bacteriophage M13 Templates Technique yields a heterogeneous population of short radiolabeled molecules 200-300 nucleotides in length. These probes are synthesized, as in Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates, by extension of an oligonucleotide primer on a single-stranded DNA template. The radiolabeled products of the reaction are then separated from the template by electrophoresis through a denaturing gel from which they are eluted directly into hybridization buffer. ...
Description of Phage Display (from New England Biolabs). Phage display describes a selection technique in which a peptide or protein is expressed as a fusion with a coat protein of a bacteriophage, resulting in display of the fused protein on the exterior surface of the phage virion, while the DNA encoding the fusion resides within the virion. Phage display has been used to create a physical linkage between a vast library of random peptide sequences to the DNA encoding each sequence, allowing rapid identification of peptide ligands for a variety of target molecules (antibodies, enzymes, cell-surface receptors, etc.) by an in vitro selection process called biopanning.. In its simplest form, biopanning is carried out by incubating a library of phage-displayed peptides with a plate (or bead) coated with the target, washing away the unbound phage, and eluting the specifically-bound phage. (Alternatively the phage can be reacted with the target in solution, followed by affinity capture of the ...
1IFI: MOLECULAR MODELS AND STRUCTURAL COMPARISONS OF NATIVE AND MUTANT CLASS I FILAMENTOUS BACTERIOPHAGES FF (FD, F1, M13), IF1 AND IKE
N ew versions of the pADL phagemid vectors pADL-20b, pADL-22b, and pADL-23b are now available. The sequence of the Amp(R) gene sequence has been modified, allowing the cloning site to be opened by either SfiI or BglI restriction enzyme. This modification results in lower background during cloning and brings more flexibility for designing libraries. The pADL vector series is a collection of phagemid vectors for controlled display on the minor coat protein III which combines HIS tag for purification, epitope tag for detection and amber stop codon for conditional display. Expression of the fusion protein is tightly controlled, eliminating issues with toxic clones and library misrepresentation.. ...
TY - JOUR. T1 - Identification of peptide mimotopes of gp96 using single-chain antibody library. AU - Shanmugam, Arulkumaran. AU - Suriano, Robert. AU - Goswami, Neha. AU - Chaudhuri, Devyani. AU - Ashok, Badithe T.. AU - Rajoria, Shilpi. AU - George, Andrea L.. AU - Mittelman, Abraham. AU - Tiwari, Raj K.. N1 - Funding Information: Acknowledgments We gratefully acknowledge the financial assistance of AVT and the National Cancer Institute RO1 grant 1RO1CA131946 (RKT).. PY - 2011/3. Y1 - 2011/3. N2 - Heat shock proteins such as gp96 are immunogenic and are widely used as vaccines in immunotherapy of cancers. The present study focuses on the use of peptide mimotopes as immunotherapeutic vaccines for prostate cancer. To this end, we developed a 15-mer gp96 peptide mimotope specifically reactive to MAT-LyLu gp96-peptide complex using combinatorial single-chain antibody and peptide phage display library. The immunogenicity of the synthesized gp96 mimotope was analyzed initially in normal BALB/c mice ...
The aim of this work was to verify the possibility of applying affinity chromatography in bacteriophage purification, from the perspective of therapeutic purposes. Elution profiles of phages modified with specific affinity motifs (Figures 3,4,5 and 6) show substantially higher phage concentration in elution fractions compared to final washing samples. This indicates binding of modified phages to the affinity resins and effective elution with standard competitive agents. Thus, affinity tags can be successfully incorporated into the T4 phage capsid by the in vivo phage display technique and they strongly elevate bacteriophage affinity to a specific resin. Non-specific binding was also observed: unmodified phages or those modified with the non-specific tag were eluted with the titre 104-105 pfu/ml. Nevertheless, the unspecific binding is 102-105 times weaker than the specific one and importantly it does not interfere with the aim of preparation of purified anti-bacterial active bacteriophages for ...
A phagemid or phasmid is a plasmid that contains an f1 origin of replication from an f1 phage. It can be used as a type of cloning vector in combination with filamentous phage M13. A phagemid can be replicated as a plasmid, and also be packaged as single stranded DNA in viral particles. Phagemids contain an origin of replication (ori) for double stranded replication, as well as an f1 ori to enable single stranded replication and packaging into phage particles. Many commonly used plasmids contain an f1 ori and are thus phagemids. Similarly to a plasmid, a phagemid can be used to clone DNA fragments and be introduced into a bacterial host by a range of techniques, such as transformation and electroporation. However, infection of a bacterial host containing a phagemid with a helper phage, for example VCSM13 or M13K07, provides the necessary viral components to enable single stranded DNA replication and packaging of the phagemid DNA into phage particles. The helper phage infects the bacterial ...
During PACE, host E. coli cells flow continuously into a fixed-volume vessel (a lagoon) containing a population of filamentous bacteriophages that encode a library of evolving proteins. The lagoon is continuously drained to a waste container after passing through an in-line luminescence monitor that measures expression from a gene III-luciferase cassette on the AP. Dilution occurs faster than cell division but slower than phage replication. Each phage carries a protein-encoding gene to be evolved instead of a phage gene (gene III) that is required for infection. Phage encoding active library members trigger expression of gene III on the AP in proportion to the desired activity and consequently produce infectious progeny. Phage encoding less active library members produce fewer infectious progeny and are lost by dilution. From: Nat. Chem. Biol. 10, 216-222 (2014). PDF. ...
mAbs. Col-1, an immunoglobulin G (IgG)-2a mAb, was purchased from Zymed. The isotype control IgG2a was obtained from Sigma and used as a control antibody.. Phage library and biopanning. Three successive rounds of biopanning with the anti-CEA antibody Col-1 were done with two pIII-display peptide phage libraries: CL10 and LL9. CL10 expresses cysteine-flanked decapeptides circularized by disulfide bridges and LL9 comprises nine linear peptides. The libraries were applied separately or pooled for biopannings. They were kindly provided by Prof. Dr. Luca Mazzucchelli (University of Bern, Bern, Switzerland; ref. 18). Biopannings were done as outlined in ref. 19, with slight modifications. In short, ELISA plates (Nunc) were coated with 40 μg/mL Col-1 in bicarbonate buffer (pH 8.5). Wells were blocked with PBS/1% dry milk and incubated with an aliquot of the phage library (∼5 × 1010 phage particles) in PBS/1% dry milk/0.1% Tween 20 at 37°C and 4°C for 1 h each. Unbound phage particles were removed ...
Tupac Shakur Ratha Be Ya Nigga lyrics & video : (feat. Richie Rich) [Intro: Richie Rich, 2Pac] [RR] Pac [PAC] Hey [RR] Whats happenin [PAC] Not motherf**kin double R, Richie R...
TY - JOUR. T1 - Automated panning and screening procedure on microplates for antibody generation from phage display libraries. AU - Turunen, Laura. AU - Takkinen, Kristiina. AU - Söderlund, Hans. AU - Pulli, Timo. PY - 2009. Y1 - 2009. N2 - Antibody phage display technology is well established and widely used for selecting specific antibodies against desired targets. Using conventional manual methods, it is laborious to perform multiple selections with different antigens simultaneously. Furthermore, manual screening of the positive clones requires much effort. The authors describe optimized and automated procedures of these processes using a magnetic bead processor for the selection and a robotic station for the screening step. Both steps are performed in a 96-well microplate format. In addition, adopting the antibody phage display technology to automated platform polyethylene glycol precipitation of the enriched phage pool was unnecessary. For screening, an enzyme-linked immunosorbent assay ...
Transposable elements have emerged as a promising candidate for human non-viral gene-therapy. The Tc1/mariner transposon Sleeping Beauty is to date one of the most efficient transposons in mammals. Sleeping Beauty transposase has so far mostly been delivered to cells via a DNA source. This might cause spontaneous integration of the transposase gene and cause fatal damage to the affected cell. Hence, it would be advantageous to employ a non-genetic source for the transposase. We here show that a novel Cell-penetrating peptide, M918, has the ability to facilitate cellular delivery of both the transposase Sleeping Beauty as a protein and a transposon donor-plasmid carrying an antibiotic resistance gene in vitro. The technique is a simple and straightforward one-step method that might render a safe and efficient delivery platform for Sleeping Beauty mediated gene therapy.. ...
miR-6883 Family miRNAs Target CDK4/6 to Induce G1 Phase Cell Cycle Arrest in Colon Cancer Cells Ectopic expression of miR-6883-5p or miR-149* downregulated CDK4 and CDK6 levels in human colorectal cancer cells. [Cancer Res] Abstract USP7 Is a Tumor-Specific WNT Activator for APC-Mutated Colorectal Cancer by Mediating β-Catenin Deubiquitination Researchers found that the β-catenin inhibitory domain (CID) in adenomatous polyposis coli (APC) represents the threshold for pathological levels of Wnt activation and tumor transformation. Mechanistically, CID-deleted APC truncation promotes β-catenin deubiquitination through reverse binding of β-TrCP and USP7 to the destruction complex. [Cell Rep] Abstract , Graphical Abstract , Press Release Identification of a Specific Peptide Binding to Colon Cancer Cells from a Phage-Displayed Peptide Library A peptide termed as CBP-DWS, which was demonstrated to be capable of binding to a panel of human colon cancer cell lines and tissues, was identified; it had ...
Phage display has become an established, widely used method for selection of peptides, antibodies or alternative scaffolds. The use of phage display for the selection of antigens from genomic or cDNA libraries of pathogens which is an alternative to the classical way of identifying immunogenic proteins is not well-known. In recent years several new applications for oligopeptide phage display in disease related fields have been developed which has led to the identification of various new antigens. These novel identified immunogenic proteins provide new insights into host pathogen interactions and can be used for the development of new diagnostic tests and vaccines. In this review we focus on the M13 oligopeptide phage display system for pathogen research but will also give examples for lambda phage display and for applications in other disease related fields. In addition, a detailed technical work flow for the identification of immunogenic oligopeptides using the pHORF system is given. The described
Culture colony of the bacteriophage M13, used as a source of DNA fragments containing the Sry (sex- determining region Y) gene, the switch that redirects mammalian development from its usual pathway - female - to male. Joint research by the Medical Research Council & Imperial Cancer Research Fund showed that some normal, chromosomally-female mice embryos developed as males with testes following their injection with a small 14-kilobase fragment of DNA containing Sry. The DNA for Sry was released from its phage vector by digestion with a restriction enzyme, isolated by gel electrophoresis (part of the autoradiogram is shown here) & then purified. - Stock Image G210/0328
Described and claimed herein are combinatorial synthetic Fab libraries displayed on a phage pIX protein. The libraries were built on scaffolds representing the most frequently used genes in human anti
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In article ,tyr-2-0706971255440001 at news.srv.ualberta.ca,, tyr-2 at bones.biochem.ualberta.ca (Karl Fischer) wrote: , In article ,hgtsei-0706971555000001 at ukmhg03.med-hg.uni-sb.de,, , hgtsei at med-rz.uni-sb.de (Thomas Seib) wrote: , , ,snip, , , Hello Thomas, , , I had similar problems with plaquing when I first started (though I was , not working with phage display). A few things I found were important were: , , 1) The selection of host. , , I started using an aliquot of JM109 which came with a kit and found that , infection with R408 or K07 was pretty pathetic; Im biased against JM109 , cause in the past it has always been a very sickly bug to work with. , , I switched to JM103 and things improved markedly. , , 2) The growth of host. , , I found that I could get consistent plaque production if I grew a colony , of JM103 overnight in 5ml of M9 medium (with CaCl2 and MgSO4)-0.6% , glycerol-0.2% casamino acids-0.002% thiamine at 37C. , , 3) Infection of host. , , I make dilutions of phage ...
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558117PRTArtificial Sequenceisolated from phage display libraries 1Ser Ser His Lys His Pro Val Thr Pro Arg Phe Phe Val Val Glu Ser 1 5 10 15Arg222PRTArtificial Sequenceisolated from phage display libraries 2Ser Ser Cys Asn Cys Tyr Val Thr Pro Asn Leu Leu Lys His Lys Cys 1 5 10 15Tyr Lys Ile Cys Ser Arg 20322PRTArtificial Sequenceisolated from phage display libraries 3Ser Ser Cys Ser His Asn His His Lys Leu Thr Ala Lys His Gln Val 1 5 10 15Ala His Lys Cys Ser Arg 20422PRTArtificial Sequenceisolated from phage display libraries 4Ser Ser Cys Asp Gln Asn Asp Ile Phe Tyr Thr Ser Lys Lys Ser His 1 5 10 15Lys Ser His Cys Ser Arg 20522PRTArtificial Sequenceisolated from phage display libraries 5Ser Ser Ser Ser Asp Val Tyr Leu Val Ser His Lys His His Leu Thr 1 5 10 15Arg His Asn Ser Ser Arg 20622PRTArtificial Sequenceisolated from phage display libraries 6Ser Ser Ser Asp Lys Cys His Lys His Trp Tyr Cys Tyr Glu Ser Lys 1 5 10 15Tyr Gly Gly Ser Ser Arg 20714PRTArtificial Sequenceisolated from phage display ...
Dr. Scott received her PhD for work on the germline immunoglobulin V genes. She attended medical school with the goal of becoming an academic biomedical researcher. Her postdoctoral research included projects to analyze the spectra of mutational hot-spots (W.G. Thilly), the development of the first phage-displayed peptide libraries and their use in analyzing antibody specificity (G.P. Smith) and in developing peptide mimics of a discontinuous protein epitope (E.D. Getzoff & J.A. Tainer). She began working at SFU in 1993 as an Assistant Professor in the Dept. of Chemistry and Member of the Institute of Molecular Biology & Biochemistry at SFU. (The Institute became a Department in SFUs Faculty of Science in 2001.) Dr. Scott was promoted to Associate Professor in 1998 and to Professor in 2002. In 2004, Dr. Scott began a joint appointment in the newly-formed Faculty of Health Sciences, as one of its founding faculty members. That year, she was awarded a Canada Research Chair in Molecular ...
Phage display technology for identifying selective antigens and address molecules on brain endothelial cells. In Blood Barrier: Biology and Research Protocols (Ed. S. Nag)
Rx Biosciences provides quality custom phage display library construction and screening services for biological research and drug discovery related projects. The gene of interest is randomly mutated to produce various combinations of the peptides or small antibodies [e.g. scFv and Fab] or proteins which get displayed on the surface of a filamentous phage [M13, fd, and f1 strains] as fusion proteins. Using a binding affinity-based process called panning; a small number of phages that display proteins specifically binding to a target of interest are recovered from the phage library that usually has a repertoire of many billions of unique displayed proteins. Finally, the proteins displayed by the selected phages are identified by phage amplification followed by DNA sequencing.. ...
A type of single-stranded DNA bacteriophage ( virus which infects bacteria ) that has a capsid which is long and thin, like a filament. Examples include the viruses F1 and M13 ...
CAR-T cell therapy emerged in the last years as a great promise to cancer treatment. Nowadays, there is a run to improve the breadth of its use, and thus, new chimeric antigen receptors (CAR) are...
Creative Biolabs is one of the well-recognized expert who is professional in applying various phage display technologies to offer library construction and screening related services for a broad range of project objectives.
Every day, headlines detail the casualties of the nations surge in heroin and prescription painkiller abuse: the funerals , the broken families and the
The fitness landscape in sequence space determines the process of biomolecular evolution. To plot the fitness landscape of protein function, we carried out in vitro molecular evolution beginning with a defective fd phage carrying a random polypeptide of 139 amino acids in place of the g3p minor coat protein D2 domain, which is essential for phage infection. After 20 cycles of random substitution at sites 12-130 of the initial random polypeptide and selection for infectivity, the selected phage showed a 1.7×10^4-fold increase in infectivity, defined as the number of infected cells per ml of phage suspension. Fitness was defined as the logarithm of infectivity, and we analyzed (1) the dependence of stationary fitness on library size, which increased gradually, and (2) the time course of changes in fitness in transitional phases, based on an original theory regarding the evolutionary dynamics in Kauffmans n-k fitness landscape model. In the landscape model, single mutations at single sites among ...
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.
A helper phage technology (Hyperphage System) was developed by Rondot et al. (Nature Biotechnology 19:75-81, 2001). The Hyperphage System allows to improve antibody presentation in phage display by increasing the number of antibodies displayed per phage particle (up to 5 vs. 0.01) and thereby the system offers great advantage in the fields of functional gene analysis and proteomics. Panning of phages can be performed with small amounts of antigen and higher efficiency. For example, the application of universal libraries for antibody isolation can be improved by employing panning of hyperphage-packed libraries on blots of protein spots after 2-dimensional gel electrophoresis. ...
Our long-term goal is to develop a process for generating a set of tumor-binding ligands for an individual patient. The initial strategy used here was to administer a phage library i.v., surgically sample tumor tissue, and collect phage that have bound to the tumor. This work follows preclinical toxicity testing ( 32) and approval by the FDA to conduct a phase I study in human cancer patients. As a phase I study, our focus in the first three patients was on the safety of phage infusion only. In the fourth patient (180-04), we obtained a large number of clones which were sequenced and analyzed for possible motif sharing among them. Later in other two patients with sufficient preinfusion tumor tissue, we conducted clone binding studies. We present here our preliminary clinical data showing (i) safety of phage display library administration; (ii) recovery of phage clones from patient tumors; (iii) identification of tumor-binding clones; (iv) cell-specificity of tumor-binding clones; and (v) ...
P hagemid vectors pADL-20c, pADL-22c, and pADL-23c are now available in store. To get full advantage of the amber suppression for expression of soluble antibodies and direct detection without sub-cloning, we adjusted the level of expression to match the high expression vector pCOMB3, a popular phagemid known for its robust display and moderate toxicity. A transcription terminator has been added after gene III to prevent excessive production of beta-lactamase during induction period. ...
A display system includes a printed display formed on a substrate and a printed battery in electrical communication with the printed display. The printed display provides power to the printed display. Since both the display and battery are printed, the resulting display system is extremely thin and the manufacture thereof is reliable and inexpensive. The display system contemplates various types of printed displays such as an electrochromic display, a thermochromic display, an electroluminescent display, or an electrophoretic display.
Microbiology students of [http://www.biology.ucsd.edu/classes/bimm120.SP07/ Kit Pogliano and Rachel Larsen] at the University of California at San Diego are currently authoring the following pages as term projects for their course, BIMM120 Bacteriology.,br> ,table border=0 cellpadding=2 cellspacing=2 width=100%> ,tr valign=top> ,td> [[Acidothermus cellulolyticus]],br> By Chau Nguyen [[Acinetobacter baumannii]],br> By Jennifer Wang [[Actinobacillus pleuropneumoniae]],br> By Nhu Le [[Agrobacterium tumefaciens]],br> By Allan Wu [[Alcanivorax borkumensis]],br> By Justin Shih [[Anaeromyxobacter dehalogenans]],br> By Lori Nacario [[Anaplasma marginale]],br> By Patricia Shih [[Archaeoglobus fulgidus]],br> By Mike Sha [[Azoarcus sp. strain BH72]],br> By Caine Ballew [[Bacillus anthracis]], causes anthrax By Grace Ucar [[Bacillus clausii]],br> By Ankur Pate [[Bacillus ...
The most often used are genes pIII or pVIII of bacteriophage M13. The next step is the capturing step. It involves conjugating ... This involves inserting foreign desired gene segments into a region of the bacteriophage genome, so that the peptide product ... It utilizes the binding interactions so that only specific peptides presented by bacteriophage are bound to the target. For ... The end result is the peptides produced by bacteriophage are specific. The resulting filamentous phages can infect gram- ...
The process involves the folding of a long single strand of viral DNA (typically the 7,249 bp genomic DNA of M13 bacteriophage ...
South Africa M13, the Mathieu groupoid by John Horton Conway M13 bacteriophage, a virus that infects bacteria Magic 2013, the ... M13, M-13 or M13 may refer to: Fiat M13/40, an Italian tank used in World War II M13 Half-track, a U.S. anti-aircraft gun used ... in World War II M13 link, a machine gun's ammunition link M-13 rocket, a version of the Soviet World War II RS-82 rocket M13 ... a state highway in the United States M13 (Cape Town), a metropolitan road near Cape Town, South Africa M13 (Durban), a ...
M13 Leviviridae. Nonenveloped, isometric. Linear ssRNA. MS2, Qβ Microviridae. Nonenveloped, isometric. Circular ssDNA. ΦX174 ... Bacteriophages are among the most common and diverse entities in the biosphere.[1] Bacteriophages are ubiquitous viruses, found ... A bacteriophage (/bækˈtɪərioʊfeɪdʒ/), also known informally as a phage (/feɪdʒ/), is a virus that infects and replicates within ... 2×108 bacteriophages per mL.[47] Bacteriophages are thought to extensively contribute to horizontal gene transfer in natural ...
Geminiviruses replicate via a rolling circle mechanism like bacteriophages such as M13, and many plasmids. Replication occurs ...
Genetically engineered M13 bacteriophage viruses allow preparation of quantum dot biocomposite structures.[23] It had ... This system allowed them to vary both the length of bacteriophage and the type of inorganic material through genetic ... M13, and TMV) are adjustable by controlling the solution concentrations, solution ionic strength, and the external magnetic ...
Bacteriophage Vectors are insertions carried by the bacteriophage λ genome can accommodate up to 12 kbp without disrupting the ... Yanisch-Perron, Celeste; Vieira, Jeffrey; Messing, Joachim (1985-01-01). "Improved M13 phage cloning vectors and host strains: ... that consist of a multiple cloning site and allow for more efficient sequencing and cloning using a set of universal M13 ...
The major coat protein of bacteriophage M13 is synthesized as a precursor, the procoat, with a typical leader (signal) sequence ... Bacteriophage M13 procoat protein inserts into the plasma membrane as a loop structure ... Bacteriophage M13 procoat protein inserts into the plasma membrane as a loop structure ... Bacteriophage M13 procoat protein inserts into the plasma membrane as a loop structure ...
With a mass of ~1.6 x 107 Daltons and com- posed of approximately 2700 proteins, bacteriophage M13 has been employed as a ... Taylor Urquhart, Elisabeth Daub, John Honek (2016). Bioorthogonal Modification of the Major Sheath Protein of Bacteriophage M13 ... Bioorthogonal Modification of the Major Sheath Protein of Bacteriophage M13: Extending the Versatility of Bionanomaterial ... conditions were developed to produce M13 bacteri- ophage labeled with over 350 active azides (estimated by fluorescent dye ...
High impact information on Bacteriophage M13. *A M13 phage peptide library was next constructed and screened with DR1 molecules ... Gene context of Bacteriophage M13. *By monovalently displaying human LIF on the surface of M13 phage and randomizing clusters ... Analytical, diagnostic and therapeutic context of Bacteriophage M13. *The wide type M13 phage, 3 HVR region of alpha globin ... Chemical compound and disease context of Bacteriophage M13. *We then used a library of random heptapeptides displayed at the ...
Capsid model of M13 bacteriophage virus from Magic-angle spinning NMR and Rosetta modeling ... Capsid model of M13 bacteriophage virus from Magic-angle spinning NMR and Rosetta modeling. *DOI: 10.2210/pdb2MJZ/pdb ... The NMR-Rosetta capsid model of M13 bacteriophage reveals a quadrupled hydrophobic packing epitope.. Morag, O., Sgourakis, N.G. ... Here we present a structural model for the capsid of intact M13 bacteriophage using Rosetta model building guided by structure ...
Temperate bacteriophage of the genus INOVIRUS which infects enterobacteria, especially E. coli. It is a filamentous phage ... Bacteriophage M13. Known as: Enterobacteria phage M13, Coliphage M13, Phages, M13 (More). ... The leader peptide of bacteriophage M13 procoat inhibited the cleavage of M13 procoat or pre-maltose-binding protein by ... Filamentous bacteriophage M13 is a single-stranded DNA phage about 65 A in diameter and 9300 A long. X-ray diffraction studies ...
... of a genetically-modified gold-binding M13 bacteriophage was investigated as a scaffold for gold synthesis. Repeated mixing of ... M13 bacteriophage spheroids as scaffolds for directed synthesis of spiky gold nanostructures T. Ngo-Duc, J. M. Plank, G. Chen, ... M13 bacteriophage spheroids as scaffolds for directed synthesis of spiky gold nanostructures† ... The spherical form (s-form) of a genetically-modified gold-binding M13 bacteriophage was investigated as a scaffold for gold ...
Biocatalytically induced surface modification of the tobacco mosaic virus and the bacteriophage M13 V. Vignali, B. S. Miranda, ... The tobacco mosaic virus (TMV) and the bacteriophage M13 have been successfully modified via laccase induced free radical ... Biocatalytically induced surface modification of the tobacco mosaic virus and the bacteriophage M13† ...
MOLECULAR MODELS AND STRUCTURAL COMPARISONS OF NATIVE AND MUTANT CLASS I FILAMENTOUS BACTERIOPHAGES FF (FD, F1, M13), IF1 AND ... We describe molecular models of the class I filamentous bacteriophages. This class includes strains fd, f1, M13 (these 3 very ... MOLECULAR MODELS AND STRUCTURAL COMPARISONS OF NATIVE AND MUTANT CLASS I FILAMENTOUS BACTERIOPHAGES FF (FD, F1, M13), IF1 AND ... Molecular models and structural comparisons of native and mutant class I filamentous bacteriophages Ff (fd, f1, M13), If1 and ...
Protocol 9: Plating Bacteriophage M13 *Protocol 10: Growing Bacteriophage M13 in Liquid Culture *Protocol 11: Preparation of ... Growing Bacteriophage M13 in Liquid Culture. (Protocol summary only for purposes of this preview site). Stocks of bacteriophage ... Protocol 8: Preparation of Single-Stranded Bacteriophage M13 DNA by Precipitation with Polyethylene Glycol * ... The inoculum of bacteriophage is almost always a freshly picked plaque or a suspension of bacteriophage particles obtained from ...
Natural bacteriophages T4 and M13 down-regulates Hsp90 gene expression in human prostate cancer cells (PC-3) representing a ... and bacteriophage M13 indicated by white arrows (B). Considering the small size of phage M13, we have pointed them outside the ... Natural bacteriophages T4 and M13 down-regulates Hsp90 gene expression in human prostate cancer cells (PC-3) representing a ... Natural bacteriophages T4 and M13 down-regulates Hsp90 gene expression in human prostate cancer cells (PC-3) representing a ...
3-Dimensional Structure of the Single-Stranded DNA-Binding Protein Encoded by Gene-V of the Filamentous Bacteriophage-M13 and a ...
Fibrillar M13 bacteriophages were used as basic building blocks to generate thin films with aligned nanogrooves, which, upon ... Fibrillar M13 bacteriophages were used as basic building blocks to generate thin films with aligned nanogrooves, which, upon ... Oriented Cell Growth on Self-Assembled Bacteriophage M13 Thin Films. Chemical Communications, 41(5185-5187). https://doi.org/ ...
The aim of this study was to evaluate the effects of M13 phage along with RGD peptide motif on in vitro and in vivo ... and cells attachment to M13 phage-RGD coated surface. In addition, the expression of Vascular Endothelial Growth Factor A (VEGF ... transcripts were significantly upregulated due to the HUVECs culturing on M13 phage-RGD coated surface. Furthermore, VEGF ... production were significantly increased in cells cultured on M13 phage-RGD coated surface. ...
The most often used are genes pIII or pVIII of bacteriophage M13. The next step is the capturing step. It involves conjugating ... This involves inserting foreign desired gene segments into a region of the bacteriophage genome, so that the peptide product ... It utilizes the binding interactions so that only specific peptides presented by bacteriophage are bound to the target. For ... The end result is the peptides produced by bacteriophage are specific. The resulting filamentous phages can infect gram- ...
... a single bacteriophage may have been imaged at 1720 1/cm (C=O) and 1667 1/cm (amide I) via PiFM. ... Given the height of the bacteriophage M13 (3-4 nm), ... Bacteriophage M13 and Plasmid DNA pUC1. Bacteriophage M13 and ... Given the height of the bacteriophage M13 (3-4 nm), a single bacteriophage may have been imaged at 1720 cm-1 (C=O) and 1667 cm- ...
... phage coat protein of the M13 bacteriophage widely used in the preparation of phage-display immunoglobulin l… ... strong,Mouse anti M13 Bacteriophage antibody, clone E1,/strong, recognizes the gp3 (G3P) ... Mouse anti M13 Bacteriophage antibody, clone E1 recognizes the gp3 (G3P) phage coat protein of the M13 bacteriophage widely ... References for M13 Bacteriophage antibody. * Jaye, D.L.et al. (2003) Use of real-time polymerase chain reaction to identify ...
Targeting Bacteriophage M13 with Engineered CRISPR Spacer.. Previously, Brouns et al. (17) showed that a plasmid containing an ... CRISPR/Cas mediated restriction of bacteriophage M13 requires intact PAM and PAM-proximal part of the protospacer. (A) An ... Selection of Natural M13 Mutants That Escape CRISPR/Cas Mediated Interference.. Despite efficient protection from M13 infection ... and an F-factor encoding pili that are required for M13 adsorption. The efficiency of plaquing (e.o.p.) of wild-type M13 on ...
T2 - Synthesis of procoat protein in Escherichia coli infected with bacteriophage M13 ... Synthesis of procoat protein in Escherichia coli infected with bacteriophage M13. / Ito, K.; Mandel, Gail; Wickner, W. ... Synthesis of procoat protein in Escherichia coli infected with bacteriophage M13. In: Proceedings of the National Academy of ... Synthesis of procoat protein in Escherichia coli infected with bacteriophage M13",. author = "K. Ito and Gail Mandel and W. ...
A simple M13 bacteriophage-based colorimetric sensor was fabricated by a simple pulling technique, and M13 bacteriophage was ... Color sensor systems based on M13 bacteriophage are being considerably researched. Although many studies on M13 bacteriophage- ... chemical sensing properties of M13 bacteriophage-based nanostructures should result in wider applications of M13 bacteriophage- ... J.-S. Moon, M. Park, W.-G. Kim, C. Kim, J. Hwang, D. Seol, C.-S. Kim, J.-R. Sohn, H. Chung, and J.-W. Oh, "M-13 bacteriophage ...
The M13 bacteriophage (M13 phage) has merged as a novel organic building block in biomedical applications. M13 phage is a ... The decoration of GO and RGD peptide was readily achieved by using RGD peptide-displaying M13 bacteriophage (RGD-M13 phage) and ... RGD peptide-displaying M13 bacteriophage/PLGA nanofibers as cell-adhesive matrices for smooth muscle cells. J Korean Phys Soc. ... Cell-adhesive RGD peptide-displaying M13 bacteriophage/PLGA nanofiber matrices for growth of fibroblasts. Biomater Res. 2014;18 ...
Searching for M13 bacteriophage with high affinity for nanodiamond particles  Au, Ho Yin (Massachusetts Institute of ...
Media in categorie "Bacteriophages". Deze categorie bevat de volgende 77 bestanden, van in totaal 77. ... ADVERTISEMENT; Antivirus and bacteriophages Wellcome L0032605.jpg 5.228 × 3.451; 7,23 MB. ... The arrangement of known genes of bacteriophage T12 after integration into host.png 917 × 456; 28 kB. ... Overgenomen van "https://commons.wikimedia.org/w/index.php?title=Category:Bacteriophages&oldid=310610186" ...
... deposited by cells cultured on aligned bacteriophage M13 thin films. Together they form a unique fingerprint. * Bacteriophages ... In our previous study, a convective assembly of bacteriophage M13 resulted in thin films which could be used to control the ... In our previous study, a convective assembly of bacteriophage M13 resulted in thin films which could be used to control the ... Visualizing cell extracellular matrix (ECM) deposited by cells cultured on aligned bacteriophage M13 thin films. / Wu, Laying; ...
1d depicts the virus kill results for bacteriophage M13.. FIG. 2 comprises two graphs depicting the results of the inactivation ... 3d depicts the virus kill results for bacteriophage M13.. FIG. 4 comprises two graphs depicting the results of the inactivation ... 1 comprises four graphs depicting the results of the inactivation by AMT and UVA of VSV and bacteriophage M13 in the presence ... 3 comprises four graphs depicting the results of the inactivation of VSV and bacteriophage M13 by AMT and UVA in the presence ...
3. Working with Bacteriophage M13 Vectors 4. Working with High-capacity Vectors ...
For M13 coat protein recombinants with DMPC alone, the relative association constants were determined for the ... Bacteriophage M13 major coat protein has been incorporated at different lipid/protein ratios in lipid bilayers consisting of ... Bacteriophage M13 major coat protein has been incorporated at different lipid/protein ratios in lipid bilayers consisting of ... Characterization of molecular selectivity of charged phospholipids for the bacteriophage M13 coat protein in lipid bilayers. ...
M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins  Hess, ... We exploit bacterial sortases to attach a variety of moieties to the capsid proteins of M13 bacteriophage. We show that pIII, ... We report a first method for using M13 bacteriophage as a multifunctional scaffold for optically imaging bacterial infections ... We demonstrate that M13 virus conjugated with hundreds of dye molecules (M13-Dye) can ... ...
... coli cell extracts and M13 genomic DNA templates bearing mutagenic lesions. The assay is based on the conversion of M13 viral ... Bacteriophage M13 / genetics* * Cell Extracts * DNA Polymerase I / metabolism * DNA Polymerase II / metabolism ... coli cell extracts and M13 genomic DNA templates bearing mutagenic lesions. The assay is based on the conversion of M13 viral ... Replication of M13 single-stranded viral DNA bearing single site-specific adducts by escherichia coli cell extracts: ...
The process involves the folding of a long single strand of viral DNA (typically the 7,249 bp genomic DNA of M13 bacteriophage ...
Bacteriophage M13 / genetics * Binding Sites * Conserved Sequence / genetics * DNA Replication* * DNA, Viral / biosynthesis ...
  • With a mass of ~1.6 x 107 Daltons and com- posed of approximately 2700 proteins, bacteriophage M13 has been employed as a molecular scaffold in bionanomaterials fabrication. (uwaterloo.ca)
  • In order to extend the versatility of M13 in this area, residue-specific unnatural amino acid incorporation was employed to successfully display azide functionalities on specific solvent-exposed positions of the pVIII major sheath protein of this bacteriophage. (uwaterloo.ca)
  • In our case, we are interested in the internal signalling and pathways associated with the direct interaction of T4 and M13 phages with prostate cancer cells (PC3). (alliedacademies.org)
  • Hence, M13 phages with the ability of fixing biochemical ligands may be useful for future studies conducted on scaffolding and regenerative medicine. (nature.com)
  • We were able to fabricate large-area multilayer smectic phage films with high degree of ordered packing and alignment via controlled self-assembly of the M13 phages. (mrs.org)
  • Bacteriophages (phages) are viruses that infect bacteria, by injecting their viral DNA or RNA into bacterial host cells. (springer.com)
  • Welcome to the Oxford Bacteriophage Conference, Phages 2013. (lpmhealthcare.com)
  • Structure of a foreign peptide displayed on the surface of bacteriophage M13. (semanticscholar.org)
  • Inhibition of purified Escherichia coli leader peptidase by the leader (signal) peptide of bacteriophage M13 procoat. (semanticscholar.org)
  • This indicates that M13 procoat inserts into the membrane as a loop structure and that the NH2-terminus of a leader peptide remains within the cytoplasm during membrane insertion. (sciencemag.org)
  • The aim of this study was to evaluate the effects of M13 phage along with RGD peptide motif on in vitro and in vivo vascularization. (nature.com)
  • Therefore, the aim of the current study was to evaluate the role of M13 phage and RGD peptide motif in stimulating angiogenic responses of in vitro cultured endothelial cells and in vivo implanted PLGA/PCL scaffolds. (nature.com)
  • This involves inserting foreign desired gene segments into a region of the bacteriophage genome, so that the peptide product will be displayed on the surface of the bacteriophage virion. (wikipedia.org)
  • The decoration of GO and RGD peptide was readily achieved by using RGD peptide-displaying M13 bacteriophage (RGD-M13 phage) and electrospinning. (ntno.org)
  • The present invention relates to vaccines comprising a bacteriophage which has been engineered to express an immunogenic protein/peptide and wherein the surface of the bacteriophage has not been modified to contain proteins/peptides designed to target the phage to receptors on the surface of specific cell types. (patentgenius.com)
  • Filamentous bacteriophage M13 is a single-stranded DNA phage about 65 A in diameter and 9300 A long. (semanticscholar.org)
  • The assay is based on the conversion of M13 viral single-stranded DNA (ssDNA) bearing a single site-specific DNA lesion to the double-stranded replicative form (RF) DNA, and permits one to quantitatively measure the efficiency of translesion synthesis. (nih.gov)
  • Filamentous bacteriophages (fd, M13, f1) contain single-stranded circular DNA of about 6400 bases (1). (springer.com)
  • The polylinker sites permitted directional cloning into single-stranded M13 bacteriophages (Medlin et al. (thefreedictionary.com)
  • Here we present a structural model for the capsid of intact M13 bacteriophage using Rosetta model building guided by structure restraints obtained from magic-angle spinning solid-state NMR experimental data. (rcsb.org)
  • We exploit bacterial sortases to attach a variety of moieties to the capsid proteins of M13 bacteriophage. (mit.edu)
  • The NMR-Rosetta capsid model of M13 bacteriophage reveals a quadrupled hydrophobic packing epitope. (semanticscholar.org)
  • The major coat protein of bacteriophage M13 is synthesized as a precursor, the procoat, with a typical leader (signal) sequence of 23 residues at its NH2-terminus. (sciencemag.org)
  • A fusion protein that contains the NH2-terminal 141 residues of cytoplasmic ribulokinase and all but the first ten residues of M13 procoat was made. (sciencemag.org)
  • Ito, K , Mandel, G & Wickner, W 1979, ' Soluble precursor of an integral membrane protein: Synthesis of procoat protein in Escherichia coli infected with bacteriophage M13 ', Proceedings of the National Academy of Sciences of the United States of America , vol. 76, no. 3, pp. 1199-1203. (elsevier.com)
  • on membrane insertion of bacteriophage M13 procoat protein, giving emphasis to structural fluctuations during assocn. (epfl.ch)
  • The process involves the folding of a long single strand of viral DNA (typically the 7,249 bp genomic DNA of M13 bacteriophage) aided by multiple smaller "staple" strands. (wikipedia.org)
  • In order to characterize mutagenic translesion DNA synthesis in UVM-induced Escherichia coli, we have developed a high-resolution DNA replication system based on E. coli cell extracts and M13 genomic DNA templates bearing mutagenic lesions. (nih.gov)
  • The efficiency of translesion synthesis across 3,N4-ethenocytosine and 1, N6-ethenoadenine is around 20%, a value that is similar to the in vivo efficiency deduced from the effect of the lesions on the survival of transfected M13 ssDNA. (nih.gov)
  • Mouse anti M13 Bacteriophage antibody, clone E1 recognizes the gp3 (G3P) phage coat protein of the M13 bacteriophage widely used in the preparation of 'phage-display' immunoglobulin libraries. (bio-rad-antibodies.com)
  • Spin-label ESR of bacteriophage M13 coat protein in mixed lipid bilayers. (ox.ac.uk)
  • Characterization of molecular selectivity of charged phospholipids for the bacteriophage M13 coat protein in lipid bilayers. (ox.ac.uk)
  • Bacteriophage M13 major coat protein has been incorporated at different lipid/protein ratios in lipid bilayers consisting of various ratios of dimyristoylphosphatidylcholine (DMPC) to dimyristoylphosphatidylglycerol (DMPG). (ox.ac.uk)
  • For M13 coat protein recombinants with DMPC alone, the relative association constants were determined for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels and found to be 1.0, 1.0, and 2.1 relative to the background DMPC, respectively. (ox.ac.uk)
  • Infection of the recombinant bacteriophage or phagemid into a specific strain of the bacterium, Escherichia coli, produced progeny phage with the coded protein displayed as a fusion with the phage's coat protein. (k-state.edu)
  • M13 phage is a long-rod shaped bacterial virus covered by ~2700 copies of α-helical pVIII coat protein. (mrs.org)
  • Spin-label electron spin resonance study of bacteriophage M13 coat protein incorporation into mixed lipid bilayers. (mpg.de)
  • Nucleotide sequence of the filamentous bacteriophage M13 DNA genome: comparison with phage fd. (semanticscholar.org)
  • He showed that the fd bacteriophage genome could be manipulated to carry a sequence of DNA coding for a protein not contained in the phage genome. (k-state.edu)
  • Along with Dr. Katarzyna Szot-Karpińska and her group at the Polish Academy of Sciences in Warsaw, they believe they are the first researchers to successfully use bacteriophages (viruses that infect and replicate within bacteria) to create a film capable of separating salt from water. (bath.ac.uk)
  • Bacteriophages, are small viruses that infect and lyse intestinal bacteria. (frontiersin.org)
  • Fibrillar M13 bacteriophages were used as basic building blocks to generate thin films with aligned nanogrooves, which, upon chemical grafting with RGD peptides, guide cell alignment and orient the cell outgrowth along defined directions. (sc.edu)
  • It utilizes the binding interactions so that only specific peptides presented by bacteriophage are bound to the target. (wikipedia.org)
  • The end result is the peptides produced by bacteriophage are specific. (wikipedia.org)
  • While different antimicrobial strategies, such as antibiotics, antimicrobial peptides, and lytic bacteriophages, offer partial solutions, what remains elusive is a generalized and programmable strategy that can distinguish between even closely related microorganisms and that allows for fine control over the composition of a microbial population. (asm.org)
  • defined growth conditions, conventional antibiotics, and antimicrobial peptides with some strain specificity, lytic bacteriophages, and the expression of antibiotic resistance genes, auxotrophic markers, or toxins under unique expression systems ( 1 ). (asm.org)
  • Belcher's virus of choice is the M13 bacteriophage, a cigar-shaped virus that replicates in bacteria. (wired.com)
  • Most likely the efficacy of probiotic bacteria has a multifactorial dependency, namely on a number of factors that include agents used, the dose, the pattern of dosing, and the characteristics of the host and the underlying luminal microbial environment and the activity of bacteriophages. (frontiersin.org)
  • A bacteriophage ( / b æ k ˈ t ɪər i oʊ f eɪ dʒ / ), also known informally as a phage ( / f eɪ dʒ / ), is a virus that infects and replicates within Bacteria and Archaea . (wikipedia.org)
  • [1] Bacteriophages are ubiquitous viruses, found wherever bacteria exist. (wikipedia.org)
  • It is estimated there are more than 10 31 bacteriophages on the planet, more than every other organism on Earth, including bacteria, combined. (wikipedia.org)
  • They found that the M13 bacteriophage, which only attacks bacteria and is benign to people, had piezoelectric properties. (electronicproducts.com)
  • That model system turned out to be a soup of saline solution containing varying concentrations of a common bacteria-attacking virus called the M13 bacteriophage. (photonics.com)
  • Additional vectors include pUC19 and pBR322, as well as bacteriophage lambda, M13 and Φ174 vectors. (neb.com)
  • Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. (nih.gov)
  • The sample contains a circular plasmid DNA pUC19 and a filamentous bacteriophage M13. (ntmdt-tips.com)
  • The spherical form (s-form) of a genetically-modified gold-binding M13 bacteriophage was investigated as a scaffold for gold synthesis. (rsc.org)
  • Arrays of structurally and genetically modified M13 bacteriophage that can determine the polarity indexes of various alcohols were found. (osapublishing.org)
  • Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. (semanticscholar.org)
  • The filamentous bacteriophages are flexible rods about 1 to 2 microns long and 6 nm in diameter, with a helical shell of protein subunits surrounding a DNA core. (rcsb.org)
  • We describe molecular models of the class I filamentous bacteriophages. (rcsb.org)
  • Although filamentous bacteriophages are known to target type IV pili, this is the first report of a phage that apparently uses a competence pilus as a receptor. (asm.org)
  • A simple M13 bacteriophage-based colorimetric sensor was fabricated by a simple pulling technique, and M13 bacteriophage was genetically engineered using a phage display technique to exhibit a negatively charged surface. (osapublishing.org)
  • M13 is widely used in phage display technology as well as in nanotechnology. (ntmdt-tips.com)
  • Unlike some other types of phage, M13 does not carry genes for toxins. (alzforum.org)
  • This antibody may be used to detect binding of ScFv antibodies developed in the M13 system. (bio-rad-antibodies.com)
  • Bacteriophage Lambda Vector for Transducing a cDNA Clone Library into Mammalian Cells", Molecular and Cellular Biology 5:1136-1142 (1985). (patentgenius.com)
  • The researchers used M13 bacteriophages, which are benign viruses, to mimic the collagen fibers in the turkey's skin. (treehugger.com)
  • The researchers applied pulses of purple-coloured light lasting just 100 femtoseconds (10 -15 seconds) to viruses called M13 bacteriophages . (newscientist.com)
  • MIT researchers used modified M13 bacteriophages as templates to assemble noble metal allow nanowires for Li-ion anode materials. (greencarcongress.com)
  • The researchers have also engineered the M13 virus to function as a scaffold to mediate the co-assembly of zinc porphyrins (photosensitizer) and iridium oxide hydrosol clusters (catalyst) for visible light-driven water oxidation. (greencarcongress.com)
  • The researchers chose the M13 virus (harmless to humans and a model organism in research labs) because its long, chopsticklike shape with a helical groove on its surface closely resembles collagen fibers. (photonics.com)
  • In order to extend the versatility of M13 in this area, residue-specific unnatural amino acid incorporation was employed to successfully display azide functionalities on specific solvent-exposed positions of the pVIII major sheath protein of this bacteriophage. (uwaterloo.ca)
  • While a phage vector limits the size of the insert into pVIII to a few amino acids, a phagemid system limits the number of copies actually displayed at the surface of M13. (stanford.edu)
  • Furthermore, VEGF protein secretion, nitric oxide, and reactive oxygen species (ROS) production were significantly increased in cells cultured on M13 phage-RGD coated surface. (nature.com)
  • We report a first method for using M13 bacteriophage as a multifunctional scaffold for optically imaging bacterial infections in vivo. (mit.edu)
  • This is an extension of the work initially reported in the journal Science in 2009 of genetically manipulating the M13 virus to support the synthesis and assembly of iron phosphate cathode materials for high-power lithium-ion batteries. (greencarcongress.com)
  • The two M13 viruses, genetically engineered for specificity (p8#9 virus) and versatility (E4/ E3 virus) served as a template for the synthesis of noble metal nanowires with diameters below 50 nm. (greencarcongress.com)
  • The inherent structural characteristic of the M13 virus enabled the synthesis of high aspect ratio nanowires. (greencarcongress.com)
  • With advantages of facile and environmentally benign synthesis, M13 biological platform proved itself as a useful toolkit for the study on the basic electrochemical property of materials. (greencarcongress.com)
  • Scanning electronic microscopic image of prostate cancer cell (PC3) interacting with bacteriophage T4 indicated by black arrows (A) and bacteriophage M13 indicated by white arrows (B). Considering the small size of phage M13, we have pointed them outside the cell surface for having clear identification and resolution. (alliedacademies.org)
  • The obtained results demonstrated the increased cellular proliferation, HUVECs migration, cells altered morphology, and cells attachment to M13 phage-RGD coated surface. (nature.com)
  • In addition, the expression of Vascular Endothelial Growth Factor A (VEGF-A), VEGF Receptors 2 and 3, Matrix Metalloproteinase 9 (MMP9), and epithelial nitric oxide synthase (eNOS) transcripts were significantly upregulated due to the HUVECs culturing on M13 phage-RGD coated surface. (nature.com)
  • Our work includes the interaction studies between different bacteriophage strains with different PCa cell lines and to know how these particles modulate cancer cell behaviour in vitro ( Figure 1 ). (alliedacademies.org)
  • We would like to take this advantage to report our preliminary findings based upon the in vitro interaction studies from PCa cells (PC3) with bacteriophage T4 and M13. (alliedacademies.org)
  • Using inductively coupled plasma-mass spectroscopy (ICP-MS), gold ion adsorption was found to be comparatively high for the gold-binding M13 spheroid, and likely contributed to the dissimilar gold morphology. (rsc.org)
  • Here we would like to shed light on "Bacteriophage" the naturally available nanoparticle, which have been effectively utilized for its anti- microbial applications for different pathogens and in other fields of biomedicine. (alliedacademies.org)
  • We also have found that bacteriophage T4 and M13 significantly downregulates Hsp90 gene ( Figure 3 ), which is involved in cellular apoptosis, as well others cellular processes, indicating bacteriophages as promising candidate nanoparticle for cancer cell therapies. (alliedacademies.org)
  • We report the development of a novel genetically-tunable piezoelectric material based on biological nanoparticle M13 bacteriophage (phage) and its application in electronic and energy generating devices. (mrs.org)
  • This class includes strains fd, f1, M13 (these 3 very similar strains are members of the Ff group), If1 and IKe. (rcsb.org)
  • For example, selecting antibody presented by bacteriophage with coated antigen in microtiter plates. (wikipedia.org)
  • Colo320 cell line stained with a cell-line specific M13 phage antibody (filled profile), compared to an irrelevant phage antibody (open profile). (bio-rad-antibodies.com)
  • The use of the s-form of the M13 bacteriophage significantly expands the templating capabilities of this viral platform and introduces the potential for further morphological control of a variety of inorganic material systems. (rsc.org)
  • The conference addressed commercial and research applications of bacteriophage in medicine, food and biotechnology, and covered a variety of topic, including pharmaceutical production and formulation, medical applications, immunogenicity and toxicology, applications in biotechnology, food technology and bacteriophage molecular biology. (lpmhealthcare.com)
  • As demonstrated in this work, this is a simple and effective method for creating a variety of structures, thus expanding the use of M13 for materials science applications and as a biological tool. (stanford.edu)
  • The bacteriophage M13 is a harmless wisp of DNA and protein about one micrometer long and 6 nanometers wide. (alzforum.org)
  • The expected contour length of the bacteriophage is ~900 nanometers. (ntmdt-tips.com)
  • The M13mp18 sequence is revised to include the G-to-T substitution in its gene II at position 6 125 bp (in M13) or 6967 bp in M13mp18. (nih.gov)
  • An MIT team including Drs. Gerbrand Ceder and Angela Belcher has synthesized gold (Au) and silver (Ag) alloy nanowires as anode materials for Li-ion batteries using multiple clones of the M13 bacteriophage virus. (greencarcongress.com)
  • protective Immune Responses Induced by the Immunization of Mice with a Recombinant Bacteriophage Displaying a Epitope of the Human Respiratory Syncytial Virus" Virology 234: 118-122 (1997). (patentgenius.com)
  • Although many studies on M13 bacteriophage-based chemical sensing of TNT, endocrine disrupting chemicals, and antibiotics have been undertaken, the fundamental physical and chemical properties of M13 bacteriophagebased nanostructures require further research. (osapublishing.org)
  • 3) M13 clones suitable for sequencing have been obtained by a new method of generating unidirectional progressive deletions from the polycloning site using exonucleases HI and VII. (nih.gov)
  • The tobacco mosaic virus (TMV) and the bacteriophage M13 have been successfully modified via laccase induced free radical formation on the tyrosine residues through single electron oxidation as the initiating step and the free radicals subsequently react with acrylate-based monomers. (rsc.org)
  • 3. The immunogenic formulation hepatitis according to claim 1, wherein the bacteriophage has been engineered to express more than one hepatitis virus type A and/or type B polypeptide. (patentgenius.com)
  • A team from the University of California, Berkeley, is apparently making use of the virus, known as M12 bacteriophage, in order to replace toxic elements that are used to charge new and old cell phones. (sellcell.com)
  • Lytic bacteriophages often offer exquisite specificity ( 2 ). (asm.org)
  • However, individual bacteriophages must be isolated against each strain and would require additional screening to determine the degree of specificity. (asm.org)