| the major mitomycin c-dna monoadduct is cytotoxic but not mutagenic in escherichia coli. | to determine the mutagenic and genotoxic properties of the major guanine n2-adduct formed by the antitumor drug mitomycin c, we have synthesized a decanucleotide, d(ttacg[mc]tatct), containing the adduct, which was inserted into a gapped bacteriophage m13 genome. analysis of the constructed genome indicated that 41% ligation of the adducted 10-mer occurred on both sides of the gap, whereas the control 10-mer ligated with 34% efficiency. after transfection of the adducted single-stranded m13 dna ... | 1998 | 9477227 |
| recombinant aspergillus fumigatus allergens: from the nucleotide sequences to clinical applications. | members of the genus aspergillus are opportunistic pathogens for cold- and warm-blooded animals. they are associated with an impressive spectrum of diseases in humans, ranging from benign colonization of the lung to life-threatening diseases such as invasive aspergillosis or allergic bronchopulmonary aspergillosis (abpa). aspergillus fumigatus is the etiological agent identified in most of the aspergillus-related human diseases and is therefore of particular clinical relevance. a major problem i ... | 1998 | 9482698 |
| uptake of foreign dna from the environment: the gastrointestinal tract and the placenta as portals of entry. | foreign dna (deoxyribonucleic acid) is part of our environment. considerable amounts of foreign dna of very different origin are ingested daily with food. in a series of experiments we fed the dna of bacteriophage m13 as test dna to mice and showed that fragments of this dna survive the passage through the gastrointestinal (gi) tract in small amounts (1-2%). food-ingested m13 dna reaches peripheral white blood cells, the spleen and liver via the intestinal epithelia and cells in the peyer's patc ... | 1998 | 9531678 |
| sec/srp-independent insertion of two thylakoid membrane proteins bearing cleavable signal peptides. | two imported thylakoid membrane proteins, psii-x and psii-w, are synthesised with cleavable n-terminal signal peptides that closely resemble those of sec-dependent lumenal proteins. in this report we have reconstituted the insertion of pre-psii-x and pre-psii-w into isolated thylakoids. we show that insertion does not require either nucleoside triphosphates or stromal extracts, both of which are required for sec- and signal recognition particle (srp)-dependent targeting mechanisms. insertion is ... | 1998 | 9537524 |
| increased misincorporation fidelity observed for nucleoside analog resistance mutations m184v and e89g in human immunodeficiency virus type 1 reverse transcriptase does not correlate with the overall error rate measured in vitro. | nucleoside analog-resistant variants of human immunodeficiency virus type 1 (hiv-1) reverse transcriptase (rt) that displayed higher in vitro polymerase fidelity were previously identified via nucleotide insertion and mispair extension assays. to evaluate the contribution of increased nucleotide insertion and primer extension fidelities on the overall error rate of hiv-1 rt, we have measured the impact of two such mutations, e89g and m184v, on dna copying fidelity in an m13 phage-based forward m ... | 1998 | 9557711 |
| virus inactivation in superoxide dismutase preparations by ultraviolet light irradiation. | viral inactivation in superoxide dismutase (sod) derived from human red cells was carried out by ultraviolet light c (uvc) irradiation. with 400 j/m2 uvc irradiation, the titer of canine parvovirus (cpv, a nonenveloped virus), m13 bacteriophage (m13, a nonenveloped phage) and vesicular stomatitis virus (vsv, an enveloped virus), which were spiked into sod solution, were reduced by > 4.6 log10 (detection limit), 7.0 log10 and 6.2 log10, respectively. the sod activity was maintained and the band p ... | 1998 | 9657049 |
| mimicking initial interactions of bacteriophage m13 coat protein disassembly in model membrane systems. | the structure and changes in environment of the m13 major coat protein were studied in model systems, mimicking the initial molecular process of the phage disassembly. for this purpose we have systematically studied protein associations with various detergents and lipids in two different coat protein assemblies: phage particles and s-forms. it is remarkable that the major coat protein can change its conformation to accommodate three distinctly different environments: phage filament, s-form, and ... | 1998 | 9665724 |
| multifunctional g3p-peptide tag for current phage display systems. | we have previously described a monoclonal antibody (mab), 10c3, directed against the gene-3 protein (g3p) of filamentous phage m13, which was produced to study g3p fusion protein expression in escherichia coli and its incorporation in the phage capsid [tesar, m., beckmann, c., röttgen, p., haase, b., faude, u., timmis, k., 1995. monoclonal antibody against piii of filamentous phage: an immunological tool to study piii fusion protein expression in phage display systems. immunology 1, 53-54]. in t ... | 1998 | 9672201 |
| cyclic peptides as non-carboxyl-terminal ligands of syntrophin pdz domains. | syntrophins, a family of intracellular peripheral membrane proteins of the dystrophin-associated protein complex (dapc), each contain a single pdz domain. syntrophin pdz domains bind c-terminal peptide sequences with the consensus r/k-e-s/t-x-v-cooh, an interaction that mediates association of skeletal muscle sodium channels with the dapc. here, we have isolated cyclic peptide ligands for syntrophin pdz domains from a library of combinatorial peptides displayed at the n terminus of protein iii o ... | 1998 | 9705339 |
| efficient display of an hcv cdna expression library as c-terminal fusion to the capsid protein d of bacteriophage lambda. | we describe the construction and characterization of a hepatitis c virus (hcv) cdna expression library displayed as a fusion to the carboxy terminus of the capsid protein d of bacteriophage lambda. cdna inserts were obtained by tagged random-priming of the hcv genome and cloned into a lambda vector from which chimeric phage bearing both wild-type d protein and d fusion products on the capsid surface were produced. the resulting library was affinity-selected with anti-hcv human monoclonal antibod ... | 1998 | 9733645 |
| solution structure of the m13 major coat protein in detergent micelles: a basis for a model of phage assembly involving specific residues. | the three-dimensional structure of the major coat protein of bacteriophage m13, solubilized in detergent micelles, has been determined using heteronuclear multidimensional nmr and restrained molecular dynamics. the protein consists of two alpha-helices, running from residues 8 to 16 and 25 to 45, respectively. these two helices are connected by a flexible and distorted helical hinge region. the structural properties of the coat protein make it resemble a flail, in which the hydrophobic helix (re ... | 1998 | 9735296 |
| on the fate of orally ingested foreign dna in mice: chromosomal association and placental transmission to the fetus. | we have previously shown that, when administered orally to mice, bacteriophage m13 dna, as a paradigm foreign dna without homology to the mouse genome, can persist in fragmented form in the gastrointestinal tract, penetrate the intestinal wall, and reach the nuclei of leukocytes, spleen and liver cells. similar results were obtained when a plasmid containing the gene for the green fluorescent protein (pegfp-c1) was fed to mice. in spleen, the foreign dna was detected in covalent linkage to dna w ... | 1998 | 9819049 |
| characterization of a new intrabody directed against the n-terminal region of human p53. | genes encoding the rearranged immunoglobulin heavy and light chain variable regions of do-1, a monoclonal antibody directed against human p53, have been used to construct a single-chain antibody. do-1 recognizes an n-terminal epitope in the region involved in the transactivation function of p53 and the binding of mdm2. the do-1 single chain scfv expressed in the periplasm of e. coli or at the surface of the filamentous phage m13 retained the immunological specificity and affinity of the full len ... | 1998 | 9824155 |
| human platelet antigen genotyping using a fluorescent sscp technique with an automatic sequencer. | the typing of human platelet antigens (hpa) can be useful in many clinical situations such as neonatal alloimmune thrombocytopenia, post-transfusion purpura, and platelet transfusion refractoriness. the fluorescent-based single-strand conformation polymorphism (f-sscp) technique is a fast and convenient way to perform hpa genotyping. universal sequences from phage m13 were introduced at both ends of specific pcr-products by using 5'-tailed primers. a short second round of pcr with universal prim ... | 1998 | 9827917 |
| differential use of the signal recognition particle translocase targeting pathway for inner membrane protein assembly in escherichia coli. | assembly of several inner membrane proteins-leader peptidase (lep), a lep derivative (lep-inv) that inserts with an inverted topology compared with the wild-type protein, the phage m13 procoat protein, and a procoat derivative (h1-procoat) with the hydrophobic core of the signal peptide replaced by a stretch from the first transmembrane segment in lep-has been studied in vitro and in escherichia coli strains that are conditional for the expression of either the 54 homologue (ffh) or 4.5s rna, wh ... | 1998 | 9843943 |
| abnormal, error-prone bypass of photoproducts by xeroderma pigmentosum variant cell extracts results in extreme strand bias for the kinds of mutations induced by uv light. | xeroderma pigmentosum (xp) is a rare genetic disease characterized by a greatly increased susceptibility to sunlight-induced skin cancer. cells from the majority of patients are defective in nucleotide excision repair. however, cells from one set of patients, xp variants, exhibit normal repair but are abnormally slow in replicating dna containing uv photoproducts. the frequency of uv radiation-induced mutations in the xp variant cells is significantly higher than that in normal human cells. furt ... | 1999 | 9858539 |
| evolution of peptides that modulate the spectral qualities of bound, small-molecule fluorophores. | fluorophore dyes are used extensively in biomedical research to sensitively assay cellular constituents and physiology. we have created, as proof of principle, fluorophore dye binding peptides that could have applications in fluorescent dye-based approaches in vitro and in vivo. | 1998 | 9862799 |
| a phage single-stranded dna (ssdna) binding protein complements ssdna accumulation of a geminivirus and interferes with viral movement. | geminiviruses are plant viruses with circular single-stranded dna (ssdna) genomes encapsidated in double icosahedral particles. tomato leaf curl geminivirus (tolcv) requires coat protein (cp) for the accumulation of ssdna in protoplasts and in plants but not for systemic infection and symptom development in plants. in the absence of cp, infected protoplasts accumulate reduced levels of ssdna and increased amounts of double-stranded dna (dsdna), compared to accumulation in the presence of wild-ty ... | 1999 | 9882367 |
| conformational and aggregational properties of the gene 9 minor coat protein of bacteriophage m13 in membrane-mimicking systems. | the membrane-bound state of the gene 9 minor coat protein of bacteriophage m13 was studied in various membrane-mimicking systems, including organic solvents, detergent micelles, and phospholipid bilayers. for this purpose we determined the conformational and aggregational properties of the chemically synthesized protein by cd, ftir, and hpsec. the protein appears to be in a monomeric or small oligomeric alpha-helical state in tfe but adopts a mixture of alpha-helical and random structure after s ... | 1999 | 9894010 |
| characterization of cerebrospinal fluid antibody specificities in neurocysticercosis using phage display peptide library. | to identify epitopes for antibodies present in cerebrospinal fluid (csf) samples from two patients with confirmed neurocysticercosis, we used a phage peptide library that displayed random dodecapeptides as a fusion to the minor coat protein (piii) of phage m13. to increase the specificity of selection, plates coated with anti-human fc antibody were used in five rounds of biopanning. the dna inserts of 30 selected clones were determined and the deduced amino acid sequences were analyzed. sequence ... | 1999 | 10219262 |
| crystal structure of the two n-terminal domains of g3p from filamentous phage fd at 1.9 a: evidence for conformational lability. | infection of escherichia coli by filamentous bacteriophages is mediated by the minor phage coat protein g3p and involves two distinct cellular receptors, the f' pilus and the periplasmic protein tola. recently we have shown that the two receptors are contacted in a sequential manner, such that binding of tola by the n-terminal domain g3p-d1 is conditional on a primary interaction of the second g3p domain d2 with the f' pilus. in order to better understand this process, we have solved the crystal ... | 1999 | 10329170 |
| [dna-fingerprinting of representatives of bovinae subfamilies using the telomere markers (ttaggg)4]. | the (ttaggg)4 oligonucleotide homologous to telomeric tandem repeats of human chromosomes was used for the first time as a multilocus hybridization probe for the analysis of genome variability in the two genera (bos and bison) of the bovinae subfamily. dna profiles for cattle, banteng, aurochs, and bison were obtained. hybridization spectra were represented by the discrete individual- and species-specific bands characterized by codominant inheritance. for comparison, dna profiles of the same sam ... | 1999 | 10330618 |
| determination of phage antibody affinities to antigen by a microbalance sensor system. | over the past decade, phage display has maturated to be a frequently used method for the generation of monoclonal antibodies of human origin. the essential step of this method is the "biopanning" of phage carrying functional antibody fragments on their surface on an immobilized antigen. the screening of large combinatorial gene libraries with this method usually leads to a set of diverse clones specifically binding to the antigen that need to be characterized further. beside its specificity, the ... | 1999 | 10337489 |
| synthesis of a eukaryotic virus protein in a prokaryotic viral-cell system: production of the adenovirus type 2 fiber shaft fragment by a tightly regulated t7pol-m13 expression system. | the use of recombinant technology for the production of proteins of interest in biotechnology and medicine has grown immensely during the last decade. a major problem often encountered is the degradation of the recombinant product by host cell proteases. we developed a novel system based on the cloning and expression of an inducible phage t7 rna polymerase into the main intergenic region of the phage m13-ko7. after infection of permissive bacterial strains with the engineered phage, the polymera ... | 1999 | 10381082 |
| [analysis of dna polymorphism in populations of the volga-ural region using genome fingerprinting with phage m13 dna]. | the hyperpolymorphism of minisatellite dna hybridizing with dna of bacteriophage m13 was analyzed in seven turkic and finno-ugric populations from the volga-urals region. in total, hybridization revealed 80 bspri genomic dna fragments ranging in size from 1.7 to 10 kb; the average frequency of an individual fragment was 0.299 +/- 0.020. the average number of hybridization fragments per pattern (varying from 14 to 20 in different populations) and frequencies of individual fragments showed signifi ... | 1999 | 10420275 |
| isolation of fibroblast growth factor receptor binding sequences using evolved phage display libraries. | a fusion protein construct consisting of the short form of human fibroblast growth factor (fgfr) fused to the heavy chain of mouse igg1 was used to screen four phage display libraries displaying 8, 13, 38 and 43 amino acids at the amino terminus of the bacteriophage m13 gene iii minor coat protein. phage with specific fgfr binding activity were isolated from the 13, 38 and 43 mer libraries. one of the highest affinity phage clones from the 13mer library was chosen to be further evolved by oligon ... | 1999 | 10420969 |
| escherichia coli cells bearing muta, a mutant glyv trna gene, express a reca-dependent error-prone dna replication activity. | a base substitution mutation (muta) in the escherichia coli glyv trna gene potentiates asp --> gly mistranslation and confers a strong mutator phenotype that is sos independent, but requires reca, recb and recc genes. here, we demonstrate that muta cells express an error-prone dna polymerase by using an in vitro experimental system based on the conversion of phage m13 single-stranded viral dna bearing a model mutagenic lesion to the double-stranded replicative form. amplification of the newly sy ... | 1999 | 10447883 |
| [identification and characterization of strains of bacillus thuringiensis by genomic fingerprinting using biotinylated phage m13 dna]. | genomic dna of the entomopathogenic bacterium bacillus thuringiensis was analyzed by the genomic fingerprinting technique. the biotin-labeled single-stranded dna of the phage m13 was used as a marker of hypervariable sequences. a procedure for analyzing the differentiation among various bacillus thuringiensis strains was developed. characteristic patterns of fingerprints were obtained for several strains, the main representatives of subspecies that are most frequently used in the manufacture of ... | 1999 | 10505264 |
| [genetic distances and taxonomic analysis of human populations of the volga-ural region based on data on dna polymorphism]. | dna polymorphism was studied in the human diallelic loci met and d7s23 linked to the cystic fibrosis gene, diallelic locus pah (the phenylketonuria gene), polyallelic locus apob, and hypervariable dna sequences identified by means of dna fingerprinting with phage m13 dna as a probe. the obtained data were used to calculate genetic distances and perform taxonomic analysis of populations of the volga-ural region (turkic and finno-ugric ethnic groups). the dna polymorphic systems studied were demon ... | 1999 | 10519075 |
| c8-guanine adduct-induced stabilization of a -1 frame shift intermediate in a nonrepetitive dna sequence. | the mechanism of frame shift mutagenesis induced by n-(deoxyguanosin-8-yl)-1-aminopyrene, the major dna adduct formed by the carcinogen 1-nitropyrene, was investigated by thermal melting studies of a 13-mer in which the adduct was flanked by a 5' and a 3' c. compared to the unmodified 13-mer, the adduct destabilized the duplex by 4-5 kcal/mol, and the deltadeltag value remained approximately the same regardless of which base was placed opposite the adduct. in contrast, deletion of the base oppos ... | 1999 | 10529252 |
| genotypic relatedness of yeast isolates from women infected with human immunodeficiency virus and their children. | the objective of this study was to compare polymerase chain reaction (pcr) fingerprinting with other molecular typing methods as an epidemiologic tool to investigate the transmission of candida strains between hiv-positive mothers and their children. forty-nine yeast strains (including candida albicans, candida glabrata, rhodotorula rubra, candida tropicalis, candida famata, candida dubliniensis, saccharomyces cerevisiae) from 30 individuals (15 children and 15 hiv-infected mothers or accompanyi ... | 1999 | 10536430 |
| molecular characterization by pcr-fingerprinting of candida dubliniensis strains isolated from two hiv-positive patients in spain. | six candida dubliniensis isolates were recovered from two hiv-infected individuals in the course of a prospective study of recurrent oral candidosis among hiv-positive patients in spain. candida albicans strains as well as non-albicans strains were also obtained from these two patients. c. dubliniensis strains were germ-tube-positive and produced abundant chlamydospores. fingerprinting the genomic dnas of these six c. dubliniensis with the c. albicans-specific probe 27a as well as karyotyping wa ... | 1999 | 10579091 |
| the late developmental pattern of mu transposon excision is conferred by a cauliflower mosaic virus 35s -driven mura cdna in transgenic maize. | the mudr element responsible for mutator activities in maize encodes two genes, mudra and mudrb. each encodes multiple transcripts hypothesized to regulate, directly or indirectly, the unique late timing and switch in transposition mechanism during maize development. mudra, which encodes the mura transposase, is unstable in bacterial plasmids, a technical problem solved by using phage m13 as a vector to prepare dna for biolistic transformation. in transgenic maize, a single 2.7-kb mudra cdna pre ... | 2000 | 10634904 |
| hirudin display on the surface of bacteriophage m13. | hirudin was fused to the n terminus of m13 minor protein gp3 (197-406) through a linker gggs by inserting both the hirudin gene and the gp8 signal sequence into the modified phagemid vector pcantab 5v to construct pcantab 5g8-hir. the expressed fusion protein was directed by gp8 signal peptide into the periplasm and assembled to the phage particle to form the hirudin-phage. the fusion protein and fusion phage were detected with biotin-thrombin by western blotting analysis. antithrombin activity ... | 1999 | 10668132 |
| single primer polymerase chain reaction fingerprinting for pasteurella multocida isolates from laboratory rabbits. | to evaluate a rapid polymerase chain reaction (pcr) fingerprinting technique for discriminating among pasteurella multocida isolates from laboratory rabbits. | 2000 | 10714523 |
| phage display of rnase a and an improved method for purification of phages displaying rnases. | functional ribonuclease a was presented on the surface of the filamentous phage m13 by fusion to the minor coat protein. rnase activity of the fusion protein was shown by a zymogram assay. in addition, we established a modified method for preparing rnase-displaying phages without contaminating host rnases. | 2000 | 10746750 |
| peptide mimics of the m13 coat protein transmembrane segment. retention of helix-helix interaction motifs. | sequence-specific noncovalent helix-helix interactions between transmembrane (tm) segments in proteins are investigated by incorporating selected tm sequences into synthetic peptides using the construct ckkk-tm-kkk. the peptides are of suitable hydrophobicity for spontaneous membrane insertion, whereas formation of an n-terminal s-s bond can bring pairs of tm helices into proximity and promote their parallel orientation. using the propensity of the protein to undergo thermally induced alpha-heli ... | 2000 | 10747951 |
| modulation of erm methyltransferase activity by peptides derived from phage display. | combinatorial peptide display on phage m13 protein piii was used to discover peptide sequences that selectively bind to ermc' methyltransferase from bacillus subtilis. one peptide, ac-lsgviat-nh(2), inhibited methylation in vitro with a 50% inhibitory concentration of 20 microm. interestingly, the set of six peptides which inhibited ermc' stimulated ermsf, a homologous methyltransferase from streptomyces fradiae. thus, ac-lsgviat-nh(2) may not act directly at the catalytic center of ermc', but m ... | 2000 | 10858361 |
| purification and characterization of the single-strand-specific and guanylic-acid-preferential deoxyribonuclease activity of the extracellular nuclease from basidiobolus haptosporus. | an extracellular nuclease from basidiobolus haptosporus (designated as nuclease bh1) was purified to homogeneity by ammonium sulfate precipitation, heat treatment, negative adsorption on deae-cellulose, and chromatography on phenyl-sepharose followed by fplc on phenyl-superose. the overall yield was 26%. the mr of the purified enzyme, determined by gel filtration, was 41 000 whereas by sds/page (after deglycosylation) it was 30 000. it is a glycoprotein with a pi of 6.8. the optimum ph and tempe ... | 2000 | 10931196 |
| optical flow-cell multichannel immunosensor for the detection of biological warfare agents. | an automated optical flow cell multichannel immunosensor for the detection and identification of toxins, viruses and bacterial particles is presented. a solid phase elisa, based on a peroxidase label for signal generation and on fused silica capillaries as a support for immobilized antibodies, has been employed for analyte detection and identification. the sensing and signal transducing component of the sensor consists of a light-emitting diode and a photodetector. the device is fitted with thre ... | 2000 | 10945452 |
| localization and rearrangement modulation of the n-terminal arm of the membrane-bound major coat protein of bacteriophage m13. | during infection the major coat protein of the filamentous bacteriophage m13 is in the cytoplasmic membrane of the host escherichia coli. this study focuses on the configurational properties of the n-terminal part of the coat protein in the membrane-bound state. for this purpose x-cys substitutions are generated at coat protein positions 3, 7, 9, 10, 11, 12, 13, 14, 15, 17, 19, 21, 22, 23 and 24, covering the n-terminal protein part. all coat protein mutants used are successfully produced in mg ... | 2000 | 11118542 |
| role of aromatic residues at the lipid-water interface in micelle-bound bacteriophage m13 major coat protein. | analyses of transmembrane domains of proteins have revealed that aromatic residues tend to cluster at or near the lipid-water interface of the membrane. to assess protein-membrane interactions of such residues, a viable mutant library was generated of the major coat protein of bacteriophage m13 (a model single membrane-spanning protein) in which one or the other of its interfacial tyrosine residues (tyr-21 and tyr-24) is mutated. using the interfacial tryptophan (trp-26) as an intrinsic probe, b ... | 2000 | 11123944 |
| rapid identification of a tobacco mosaic virus epitope by using a coat protein gene-fragment-pviii fusion library. | this study describes the identification of the epitope recognized by the tobacco mosaic virus (tmv) coat protein (cp)-specific monoclonal antibody 29 (mab29) by displaying a cp gene-fragment library on pviii of filamentous phage m13. more than 80% of the clones isolated after one round of panning bound specifically to mab29. dna sequencing of ten randomly chosen mab29-specific clones and subsequent sequence comparison revealed a common seven amino acid epitope (elirgtg) representing amino acids ... | 2001 | 11125152 |
| rapid titration of multiple samples of filamentous bacteriophage (m13) on nitrocellulose filters. | | 2000 | 11126120 |
| profiling the immune response in patients with breast cancer by phage-displayed cdna libraries. | display on the surface of filamentous phages has been shown to be well suited for the enrichment of serum antibody-binding ligands. here, we have taken the advantage of this technology to analyze the humoral immune response in patients with cancer. the cdna repertoires from breast cancer cell lines t47d and mcf-7 were fused to the 3'-end of the filamentous phage m13 gene vi in all three reading frames. when the libraries were biopanned on rabbit polyclonal igg against the human bcl-x(l) protein, ... | 2001 | 11241275 |
| studies on the anti-mitogenic, anti-phage and hypotensive effects of several ribosome inactivating proteins. | an investigation was conducted to compare the anti-mitogenic, anti-phage and hypotensive activities of several ribosome inactivating proteins (rips) in order to ascertain whether the rips differed in their potencies in the various bioassays. agrostin, luffin and saporin elicited a dose-dependent suppression of the mitogenic response of murine splenocytes to concanavalin a. the three rips were approximately equipotent in this regard, with near maximal inhibition attained at a dose of 83 nm and ap ... | 2001 | 11255109 |
| conformation and orientation of the gene 9 minor coat protein of bacteriophage m13 in phospholipid bilayers. | the membrane-bound state of the gene 9 minor coat protein of bacteriophage m13 was studied in model membrane systems, which varied in lipid head group and lipid acyl chain composition. by using ftir spectroscopy and subsequent band analysis a quantitative analysis of the secondary structure of the protein was obtained. the secondary structure of the gene 9 protein predominantly consists of alpha-helical (67%) and turn (33%) structures. the turn structure is likely to be located c-terminally wher ... | 2001 | 11286965 |
| spontaneous insertion of gene 9 minor coat protein of bacteriophage m13 in model membranes. | gene 9 minor coat protein from bacteriophage m13 is known to be located in the inner membrane after phage infection of escherichia coli. the way of insertion of this small protein (32 amino acids) into membranes is still unknown. here we show that the protein is able to insert in monolayers. the limiting surface pressure of 35 mn/m for 1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3-phosphoglycerol lipid systems indicates that this spontaneous insertion can also occur in v ... | 2001 | 11286974 |
| penicillium marneffei: types and drug susceptibility. | the pcr fingerprints of 30 penicillium marneffei isolates from chiang rai in northern thailand and bangkok in central thailand were studied through use of single-nucleotide primers (gaca)4 and the phage m13 core sequence. discrimination of fingerprint patterns was based on differences in the number of major bands. the p. marneffei isolates were divided into four types, i.e., a, b, c, and d. type a was found in two isolates from chiang rai (6.7%). types b and c respectively were found in two (6.7 ... | 2001 | 11307592 |
| selection of novel ligands from a whole-molecule randomly mutated c5a library. | novel antagonists of the proinflammatory leukocyte chemoattractant c5a have been produced from a phage display library of whole-molecule random mutants. the cdna for the inflammatory polypeptide c5adr(74) was used as template in a pcr reaction doped with the mutagenic nucleoside triphosphates dptp [dp: 6-(2-deoxy-beta-d-ribofuranosyl)-3,4-dihydro-8h-pyrimido-[4,5-c][1,2]oxazin-7-one] and 8-oxodgtp (8-oxodg: 8-oxo-2'-deoxyguanosine) to allow the introduction of mutations in a highly controlled ma ... | 2001 | 11342716 |
| pcr fingerprinting: a convenient molecular tool to distinguish between candida dubliniensis and candida albicans. | candida dubliniensis was recently identified as a germ-tube- and chlamydospore-positive yeast, phenotypically and morphologically indistinguishable from the phylogenetically closely related yeast species c. albicans and its synonymized variant c. stellatoidea. the high similarity between these yeast species causes significant problems in the correct identification of c. dubliniensis in a standard clinical mycology laboratory. polymerase chain reaction (pcr) fingerprinting was successfully applie ... | 2001 | 11346267 |
| on the fate of plant or other foreign genes upon the uptake in food or after intramuscular injection in mice. | uptake and persistence of the dna of bacteriophage m13 and the cloned gene for the green fluorescent protein (gfp) as test genes for food-ingested dna have previously been traced from the intestinal contents, via the gut wall, peyer's patches and peripheral white blood cells to spleen and liver, and via the placenta to fetuses and newborn animals. we have now chosen a natural scenario and fed soybean leaves to mice. the distribution of the plant-specific, nucleus-encoded ribulose-1,5-bisphosphat ... | 2001 | 11361332 |
| the rescue by phage display of human fabs to gp120 hiv-1 glycoprotein using ebv transformed lymphocytes. | human hybridomas secreting monoclonal antibodies in a stable manner are difficult to develop. the main difficulties are the restricted techniques for b-cell immortalization, the low number of sensitized b cells in peripheral blood, and the impossibility, for ethical reasons, to immunize humans with most antigens. phage display has proved to be a powerful method for the generation of recombinant antibody fragments. this technology relies on the construction of recombinant fab or scfv libraries an ... | 2001 | 11395866 |
| phage display combinatorial libraries of short peptides: ligand selection for protein purification. | a library of heptapeptides displayed on the surface of filamentous phage m13 was evaluated as a potential source of affinity ligands for the purification of rhizomucor miehei lipase. two independent selection (biopanning) protocols were employed: the enzyme was either physically adsorbed on polystyrene or chemically immobilized on small magnetic beads. from screening with the polystyrene-adsorbed lipase it was found that there was a rapid enrichment of the library with "doublet" clones i.e. the ... | 2001 | 11397457 |
| in vitro selection of enzymatically active lipase variants from phage libraries using a mechanism-based inhibitor. | the 'detergent lipase' lipolase, from thermomyces lanuginosa was subjected to a combinatorial protein engineering/phage display approach with the aim of identifying new enzyme variants with improved characteristics in the presence of detergents. first it was demonstrated that wild-type lipolase could be produced in escherichia coli retaining full activity and be displayed as an active enzyme fused to coat protein 3 on e. coli phage m13. a phagemid library designed to result in approximately two ... | 2001 | 11470533 |
| genetic differentiation in eurasian populations of the postfire ascomycete daldinia loculata. | the genetic population structure of the postfire ascomycete daldinia loculata was studied to test for differentiation on a continental scale. ninety-six samples of spore families, each comprising mycelia from six to 10 spores originating from single perithecia, were sampled from one russian and six fennoscandian forest sites. allelic distribution was assayed for six nuclear gene loci by restriction enzyme analyses of polymerase chain reaction (pcr)-amplified gene fragments. in addition, the full ... | 2001 | 11472535 |
| isolation, characterization, and recovery of small peptide phage display epitopes selected against viable malignant glioma cells. | phage display techniques rely on nearly random oligonucleotide sequences inserted into the protein iii filament binding protein of an escherichia coli filamentous phage m13 to generate a library of phage that express more than 10(7) different peptides. phage that expresses a sequence having high affinity for a specific molecule, cell, or tissue can then be isolated through selective binding and recovery. selected phage cannot only be used as gene transfer vectors in themselves, but the small pep ... | 2001 | 11498772 |
| binding of phage-displayed hiv-1 tat to tar rna in the presence of cyclin t1. | the transactivator protein (tat) of the human immunodeficiency virus (hiv) is a key regulatory protein in the viral replication cycle. together with cellular cyclin t1 and an rna element (transactivation response; tar) located at the 5' end of all viral transcripts, it forms a ternary complex that ultimately enhances the expression of all viral genes. in this ternary complex, cyclin t1 interacts directly with tat and tar. the presence of cyclin t1 is essential for high tar rna affinity and speci ... | 2001 | 11549886 |
| fluorescence resonance energy transfer shows a close helix-helix distance in the transmembrane m13 procoat protein. | during the membrane insertion process the major coat protein of bacteriophage m13 assumes a conformation in which two transmembrane helices corresponding to the leader sequence and the anchor region in the mature part of the protein coming into close contact with each other. previous studies on the molecular mechanism of membrane insertion of m13 procoat protein have shown that this interaction between the two helices might drive the actual translocation process. we investigated the intramolecul ... | 2001 | 11591151 |
| functional selection of phage displayed peptides for facilitated design of fusion tags improving aqueous two-phase partitioning of recombinant proteins. | aqueous two-phase systems allow for the unequal distribution of proteins and other molecules in water-rich solutions containing phase separating polymers or surfactants. one approach to improve the partitioning properties of recombinant proteins is to produce the proteins as fused to certain peptide tags. however, the rational design of such tags has proven difficult since it involves a compromise between multivariate parameters such as partitioning properties, solvent accessibility and producti ... | 2002 | 11690690 |
| foreign dna integration--perturbations of the genome--oncogenesis. | we have been interested in the consequences of foreign dna insertion into established mammalian genomes and have initially studied this problem in adenovirus type 12 (ad12)-transformed cells or in ad12-induced hamster tumors. since integrates are frequently methylated de novo, it appears that they might be modified by an ancient defense mechanism against foreign dna. in cells transgenic for the dna of ad12 or for the dna of bacteriophage lambda, changes in cellular methylation and transcription ... | 2001 | 11708490 |
| selection of potent chymotrypsin and elastase inhibitors from m13 phage library of basic pancreatic trypsin inhibitor (bpti). | the combinatorial approach offered by phage display has proved to be powerful in obtaining novel variants of canonical inhibitors of serine proteinases that show new binding patterns. we applied this strategy to search for variants of basic pancreatic trypsin inhibitor (bpti) that would be strong inhibitors of two serine proteinases: bovine alpha-chymotrypsin and porcine pancreatic elastase. bpti only moderately inhibits the first and does not inhibit the second enzyme. a representative library ... | 2001 | 11755204 |
| [production of phage-displayed single chain variable fragments of monoclonal antibody mgb1]. | to lay a foundation for obtaining a tumor-targeting vehicle for in vivo study on diagnosis and treatment of gastric carcinoma by generating single chain variable fragments (scfv) of monoclonal antibody mgb1 directed against the cancer. | 2000 | 11798521 |
| structural characterization of bacteriophage m13 solubilization by amphiphiles. | the structural properties of bacteriophage m13 during disassembly were studied in different membrane model systems, composed of a homologue series of the detergents sodium octyl sulfate, sodium decyl sulfate, and sodium dodecyl sulfate. the structural changes during phage disruption were monitored by spin-labeled electron spin resonance (esr) and circular dichroism spectroscopy. for the purpose of esr spectroscopy the major coat protein mutants v31c and g38c were site-directed spin labeled in th ... | 2002 | 11825608 |
| selection and identification of dense granule antigen gra3 by toxoplasma gondii whole genome phage display. | toxoplasma gondii is a ubiquitous, unicellular, eukaryotic parasite with a complex intracellular life cycle capable of invading and chronically infecting a wide variety of vertebrate host species, including man. although normally opportunistic in healthy adults, it is a lethal pathogen in immunocompromised humans, particularly in aids patients. we present the application of a genomic phage display as a tool for the direct identification of antigens with potential value in diagnosis and/or as sub ... | 2002 | 11825896 |
| uptake and processing of modified bacteriophage m13 in mice: implications for phage display. | internalization and degradation of filamentous bacteriophage m13 by a specific target cell may have major consequences for the recovery of phage in in vivo biopanning of phage libraries. therefore, we investigated the pharmacokinetics and processing of native and receptor-targeted phage in mice. (35)s-radiolabeled m13 was chemically modified by conjugation of either galactose (lacm13) or succinic acid groups (sucm13) to the coat protein of the phage to stimulate uptake by galactose recognizing h ... | 2002 | 11853411 |
| analysis of sequences conferring autonomous replication in baker's yeast. | a method is presented for rapid sequencing and mapping of elements which support autonomous replication in yeast. the strategy relies on a novel phage m13 vector which allows detection of ars (autonomously replicating sequence) function in cloned fragments. deletion mapping of an ars element linked to the ho gene of saccharomyces cerevisiae has identified a 57-bp region 3' to the gene, which is essential for autonomous replication. this region shows sequence homology to other ars elements. | 1983 | 11892814 |
| [dna fingerprinting of individual species and intergeneric and interspecific hybrids of genera bos and bison, subfamily bovinae]. | genome fingerprinting with a hypervariable minisatellite sequence of phage m13 dna was used to study the genetic variation in individual species of the genera bos and bison (subfamily bovinae) and in their interspecific and intergeneric hybrids. dna fingerprints were obtained for domestic cow bos taurus primigenius, vatussy bos taurus macroceros, banteng bos javanicus, gaur bos gaurus, wisent bison bonasus, bison bison bison, and for the interspecific and intergeneric hybrids. compared with the ... | 2002 | 12018169 |
| identification of b cell epitopes of hepatitis c virus rna dependent rna polymerase. | the aim of this study was to identify the b cell epitopes of hepatitis c virus (hcv) ns5b rna dependent rna polymerase (rdrp). the truncated hcv ns5b protein ns5b-dc21 was expressed in escherichia coli and its antigenicity was confirmed by enzyme-linked immunosorbent assay (elisa) using 130 hcv-positive human sera and 15 negative sera. antibodies specific to ns5b-dc21 protein were purified by affinity chromatography using sepharose-4b coupled with the recombinant protein. a 12-mer phage displaye ... | 2002 | 12020787 |
| novel peptides that inhibit the propagation of newcastle disease virus. | a disulfide constrained random heptapeptide library displayed on filamentous bacteriophage m13 was applied to select specific ligands that interact with newcastle disease virus (ndv). a fusion phage carrying the amino acid sequence tlttkly was selected from the panning procedure. an antibody competition assay showed that the selected phage was capable of competing with the polyclonal antibodies raised against ndv for binding sites on the virus. determination of the binding affinity of this phage ... | 2002 | 12021868 |
| the effect of an agglutogen on virus infection: biotinylated filamentous phages and avidin as a model. | to address the effect of an agglutogen on virus infection, we studied the avidin-associated inhibition of infection by biotinylated m13 phages (bio-phages). microscopic observation of mixtures of bio-phages and avidin-fluorescein conjugates revealed many aggregates. even at low phage concentrations, avidin induced inhibition of infection significantly. anti-m13 phage antibody also made aggregates and inhibited the infection but in a different manner from avidin. the inhibition by avidin was at > ... | 2002 | 12044874 |
| determination of the binding epitope for anti-trichosanthin monoclonal antibody t(8)c(12). | western blot result showed that t(8)c(12), an anti-trichosanthin (tcs) monoclonal antibody, could bind to a cnbr-cleaved tcs fragment with mw of 8 kd. the epitope was located in 1-72 of the n terminal of tcs as shown by amino acid analysis. a random 6-aa peptide library cloned in piii of phage m13 was screened by t(8)c(12). after two cycles screening, 15 positive clones were randomly selected and six kinds of 6-aa sequences were determined, which showed to be highly homologous with a ser/thr-(x) ... | 1998 | 12174274 |
| cloning, expression and display of the pres domain of hepatitis b virus on filamentous bacteriophage m13. | the pres domain of hepatitis b virus (hbv) is believed to be involved in virion assembly and attachment to a hepatocyte receptor during infection. in order to study the functions of this region, we fused it to the g3p protein of bacteriophage m13 that allows the fusion protein to be displayed at the tip of the filament. the fusion protein was detected by the anti-e tag antibody on a western blot. the polypeptide in a soluble form was produced by transfecting a non-suppressor e. coli host cell wi ... | 2002 | 12186783 |
| new approaches to improve a peptide vaccine against porcine taenia solium cysticercosis. | cysticercosis caused by taenia solium frequently affects human health and rustic porciculture. cysticerci may localize in the central nervous system of humans causing neurocysticercosis, a major health problem in undeveloped countries. prevalence and intensity of this disease in pigs and humans are related to social factors (poor personal hygiene, low sanitary conditions, rustic rearing of pigs, open fecalism) and possibly to biological factors such as immunity, genetic background, and gender. t ... | 2002 | 12234527 |
| analytical model for determination of parameters of helical structures in solution by small angle scattering: comparison of reca structures by sans. | the filament structures of the self-polymers of reca proteins from escherichia coli and pseudomonas aeruginosa, their complexes with atpgammas, phage m13 single-stranded dna (ssdna) and the tertiary complexes reca::atpgammas::ssdna were compared by small angle neutron scattering. a model was developed that allowed for an analytical solution for small angle scattering on a long helical filament, making it possible to obtain the helical pitch and the mean diameter of the protein filament from the ... | 2003 | 12606054 |
| bispecific monoclonal antibodies against a viral and an enzyme: utilities in ultrasensitive virus elisa and phage display technology. | a quadroma (hybrid-hybridoma) secreting bispecific antibodies with one paratope specific for m13 bacteriophage coat protein and another paratope specific for alkaline phosphatase (ap) was developed by electro-fusion of the two parental hybridomas and selected by a fluorescence activated cell sorter (facs). the anti-phage m13/anti-ap bsmabs were purified from anti-phage m13 monospecific mab by a novel affinity method using mimetic blue a6xl as immune complexes with ap. the purified bsmabs with po ... | 2003 | 12609538 |
| protein-lipid interactions of bacteriophage m13 major coat protein. | during the past years, remarkable progress has been made in our understanding of the replication cycle of bacteriophage m13 and the molecular details that enable phage proteins to navigate in the complex environment of the host cell. with new developments in molecular membrane biology in combination with spectroscopic techniques, we are now in a position to ask how phages carry out this delicate process on a molecular level, and what sort of protein-lipid and protein-protein interactions are inv ... | 2003 | 12659940 |
| the ompl porin does not modulate redox potential in the periplasmic space of escherichia coli. | the escherichia coli dsba protein is the major oxidative catalyst in the periplasm. dartigalongue et al. (embo j., 19, 5980-5988, 2000) reported that null mutations in the ompl gene of e.coli fully suppress all phenotypes associated with dsba mutants, i.e. sensitivity to the reducing agent dithiothreitol (dtt) and the antibiotic benzylpenicillin, lack of motility, reduced alkaline phosphatase activity and mucoidy. they showed that ompl is a porin and hypothesized that ompl null mutations exert t ... | 2003 | 12660153 |
| from est to ihc: human antibody pipeline for target research. | we have developed a method for the high-level expression of expressed sequence tags (ests) as inclusion bodies in escherichia coli by c-terminal fusion to the n1-domain of g3p of filamentous phage m13. soluble fusion protein is obtained by an efficient refolding procedure. we have applied such protein preparations to the selection of human antibody fragments from phage-displayed hucal libraries. for all fusion proteins tested in this study, hucal antibodies could be generated which specifically ... | 2003 | 12667684 |
| [differentiation of closely related and distant human populations by multilocus dna fingerprinting]. | using multilocus dna fingerprinting with phage m13 dna as a probe, we have investigated a heterogeneous group of four human populations from eastern europe and northeastern asia. these populations belong to two language families: indo-european (eastern slavonic branch: russians, belarussians) and altaian (turkic branch: yakuts). the experimental results were treated by different statistical techniques: cluster analysis, multidimensional scaling, and multiple correspondence analysis. coefficients ... | 2003 | 12669420 |
| identification of gal80p-interacting proteins by saccharomyces cerevisiae whole genome phage display. | networks of interacting proteins and protein interaction maps can help in functional annotation in genome analysis projects. we present the application of genomic phage display as a tool to identify interacting proteins in saccharomyces cerevisiae. we have developed a large phagemid display library (approximately 7.7x10(7) independent clones) of sheared s. cerevisiae genomic dna (12.1 mbp genome size) fused to gene iii (lacking the n1 domain) of the filamentous phage m13. baits tagged with an n- ... | 2003 | 12706896 |
| plaque reduction test: an alternative method to assess specific antibody response to piii-displayed peptide of filamentous phage m13. | phage-displayed peptide systems have been used to identify the immunogenic epitopes and to develop the design of peptide-based or peptide-displaying phages themselves as vaccine candidates. to estimate the humoral immunity of phage-based vaccine, it is necessary to evaluate the antibody response specifically directed at the displayed peptide. enzyme-linked immunosorbent assays (elisas) and western blot analysis are commonly used for this purpose. however, using these methods, it is not easy to d ... | 2003 | 12738371 |
| sequence context strongly modulates association of polar residues in transmembrane helices. | polar residues are capable of mediating the association of membrane-embedded helices through the formation of side-chain/side-chain inter-helical hydrogen bonds. however, the extent to which native van der waals packing of the residues surrounding the polar locus can enhance, or interfere with, the interaction of polar residues has not yet been studied. we examined the propensities of four polar residues (aspartic acid, asparagine, glutamic acid, and glutamine) to promote self-association of tra ... | 2003 | 12875850 |
| ion multivalence and like-charge polyelectrolyte attraction. | it is known empirically that multivalent ions generate attractions between like-charged polyelectrolytes, with different valence requirements for different systems. how multivalent must an ion be before it can condense a given polyelectrolyte? using charge-tunable m13 virus rods and a family of artificial homologous "dumbbell" divalent ions of different sizes, we have constructed a multivalent ion-polyelectrolyte phase diagram, and find an experimentally motivated general criterion for like-char ... | 2003 | 12906514 |
| the gastrointestinal tract as the portal of entry for foreign macromolecules: fate of dna and proteins. | the gastrointestinal tract (git) of mammals is the main portal of entry for foreign dna and proteins. we have documented the fate of orally administered dna or protein in the git of the mouse. the gene for the green fluorescent protein (gfp) (4.7 kb) and the genomes of bacteriophage m13 (7.25 kb) and adenovirus type 2 (ad2; 35.9 kb) were used as test dnas. persistence of these dnas in the git was monitored by southern hybridization and fluorescent in situ hybridization (fish) or by pcr. for stud ... | 2003 | 12938039 |
| genetically modified filamentous phage as bactericidal agents: a pilot study. | to evaluate the ability of a filamentous phage encoding lethal proteins to kill bacteria without host-cell lysis. | 2003 | 12969496 |
| some physical-chemical and biological properties of the rod-shaped coliphage m13. | | 1964 | 14227037 |
| the initial steps in infection with coliphage m13. | | 1964 | 14227038 |
| virus-based toolkit for the directed synthesis of magnetic and semiconducting nanowires. | we report a virus-based scaffold for the synthesis of single-crystal zns, cds, and freestanding chemically ordered copt and fept nanowires, with the means of modifying substrate specificity through standard biological methods. peptides (selected through an evolutionary screening process) that exhibit control of composition, size, and phase during nanoparticle nucleation have been expressed on the highly ordered filamentous capsid of the m13 bacteriophage. the incorporation of specific, nucleatin ... | 2004 | 14716009 |
| the affinity of gxxxg motifs in transmembrane helix-helix interactions is modulated by long-range communication. | sequence motifs are responsible for ensuring the proper assembly of transmembrane (tm) helices in the lipid bilayer. to understand the mechanism by which the affinity of a common tm-tm interactive motif is controlled at the sequence level, we compared two well characterized gxxxg motif-containing homodimers, those formed by human erythrocyte protein glycophorin a (gpa, high-affinity dimer) and those formed by bacteriophage m13 major coat protein (mcp, low affinity dimer). in both constructs, the ... | 2004 | 14766751 |
| polar residue tagging of transmembrane peptides. | studies that focus on packing interactions between transmembrane (tm) helices in membrane proteins would greatly benefit from the ability to investigate their association and packing interactions in multi-spanning tm domains. however, the production, purification, and characterization of such units have been impeded by their high intrinsic hydrophobicity. we describe the polar tagging approach to biophysical analysis of tm segment peptides, where incorporation of polar residues of suitable type ... | 2003 | 14991677 |
| [analysis of genomic polymorphism of typical and atypical strains of the plague pathogen using polymerase chain reaction with universal primers]. | an analysis of genome polymorphism of the y. pestis strains by using the method of polymerase chain reaction (pcr) for the tandem repeats of bacteriophage m13 dna revealed a species similarity of both typical and atypical (according to diagnostic signs) plague-microbe strains. strain y. pestis a-1726 with the atypical differential-and-diagnostic properties, without the amplicon specified for y. pestis and sized 1000 b.p., was identified among 27 analyzed y. pestis strains. the amplicon profiles ... | 2004 | 15025000 |
| the role of rna polymerase sigma subunit in promoter-independent initiation of transcription. | in bacteria, initiation of transcription depends on the rna polymerase sigma subunit, which brings catalytically proficient rna polymerase core to promoters by binding to specific dna elements located upstream of the transcription start point. here, we study sigma-dependent synthesis of a transcript that is used to prime replication of the single-stranded genome of bacteriophage m13. we show that, in this system, sigma plays no role in dna recognition, which is accomplished solely through rna po ... | 2004 | 15070729 |
| genotyping and antifungal susceptibility profile of dipodascus capitatus isolates causing disseminated infection in seven hematological patients of a tertiary hospital. | seven cases of disseminated infection due to dipodascus capitatus are reported. infections occurred in a hematological unit of a tertiary hospital during a period of 5 years. five cases were refractory to antifungal therapy. antifungal susceptibility testing of seven isolates was performed, and strains were typed by pcr fingerprinting with the core sequence of phage m13 and by random amplification of polymorphic dna with two primers, ap12h and w-80a. a very short range of mics of each antifungal ... | 2004 | 15071063 |
| dna stimulates mec1-mediated phosphorylation of replication protein a. | the cellular single-stranded dna (ssdna)-binding protein replication protein a (rpa) becomes phosphorylated periodically during the normal cell cycle and also in response to dna damage. in saccharomyces cerevisiae, rpa phosphorylation requires the checkpoint protein mec1, a protein kinase homologous in structure and function to human atr. we confirm here that immunocomplexes containing a tagged version of mec1 catalyze phosphorylation of purified rpa, likely reflecting an rpa kinase activity int ... | 2004 | 15078888 |
| identification and characterization of peptides that bind to cyanovirin-n, a potent human immunodeficiency virus-inactivating protein. | cyanovirin-n (cv-n) exerts a potent human immunodeficiency virus (hiv)-inactivating activity against diverse strains of hiv by binding to the viral surface envelope glycoprotein gp120 and blocking its essential interactions with cellular receptors. based on previous thermodynamic analyses, it has been speculated that discrete protein-protein interactions might play an important ancillary role in the cv-n/gp120 binding event, in addition to the interactions of cv-n with specific oligosaccharides ... | 2004 | 15165709 |
| two-triplet cga repeats impede dna replication in bacteriophage m13 in escherichia coli. | expansion and contraction instabilities associated with cag, cgg, gaa and cga (gac) repeats propagation cause more than a dozen human genetic diseases and cancers. in this work, the propagation behavior of a bacteriophage m13 carrying a calf prochymosin cdna fragment with a (cga)2 repeat in a small hairpin forming region is reported. such a m13 derivative when propagated in escherichia coli, produces small plaques by decreasing phage yield and also mitigates the inhibition on host cell growth, c ... | 2004 | 15293942 |
| multiplexed sorting of libraries on libraries: a novel method for empirical protein design by affinity-driven phage enrichment on synthetic peptide arrays. | chemically synthesized peptide arrays on planar cellulose carriers are proposed as libraries of ligands suitable for the multiplexed simultaneous capture of peptide-specific acceptor proteins from a large randomly mutagenized library of acceptor proteins presented on bacteriophage m13 particles. this experimental set-up can be exploited to rapidly screen for individual new, distinct binding partners from two complementary libraries (two-dimensional screening). the technical feasibility of this e ... | 2004 | 15384416 |
| antigenic properties of phage displayed peptides comprising disulfide-bonded loop of the immunodominant region of hiv-1 gp41. | the hiv-1 envelope glycoprotein gp41 contains cys(x)5cys motif, which has been shown to elicit a strong antibody response in almost all hiv-1 infected individuals. this disulfide-bonded loop region is conserved in most retroviruses suggesting the existence of an essential function in virus life cycle. in this study, we displayed the peptides comprising 12 amino acids of the immunodominant loop of gp41 on the surface of m13 phage as n-terminal fusions to the minor coat protein piii and major coat ... | 2004 | 15388262 |
| purification of filamentous bacteriophage m13 by expanded bed anion exchange chromatography. | in this paper, we investigated the development of a simplified and rapid primary capture step for the recovery of m13 bacteriophage from particulate-containing feedstock. m13 bacteriophage, carrying an insert, was propagated and subsequently purified by the application of both conventional multiple steps and expanded bed anion exchange chromatography. in the conventional method, precipitation was conducted with peg/nacl, and centrifugation was also performed. in the single step expanded bed anio ... | 2004 | 15459653 |