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molecular tracking of candida albicans in a neonatal intensive care unit: long-term colonizations versus catheter-related infections.nosocomial neonatal candidiasis is a major problem in infants requiring intensive therapy. the subjects of this retrospective study were nine preterm infants admitted to the neonatal intensive care unit of the hospital central de asturias between march 1993 and august 1994. the infants were infected with or colonized by candida albicans. five patients developed c. albicans bloodstream infections. a total of 36 isolates (including isolates from catheters and parenteral nutrition) were examined fo ...19979399489
display of functional thrombin inhibitor hirudin on the surface of phage m13.a synthetic gene for hirudin was ligated into phagemid pcantab5e. this construct allows production of either soluble hirudin or phage having hirudin displayed on the surface. similarly, hirudin variants with extensions either at their n- or c-terminus were generated. the genes were expressed in their soluble form in a non-suppressor strain of e. coli. periplasmatic fractions were evaluated in standard thrombin inhibition assays. extending hirudin by a single gln residue at the n-terminus reduces ...19979434182
production of a single-chain fragment of the murine anti-idiotypic antibody aca125 as phage-displayed and soluble antibody by recombinant phage antibody technique.the f(ab')2 fragment of the murine monoclonal anti-idiotypic antibody aca125 mimicking the tumor-associated antigen ca125 is used as a vaccine for the induction of an anti-tumoral immunity in patients with ovarian carcinoma. we tried to generate a single-chain fragment (scfv) composed of aca125 heavy- and light-chain variable domains connected by a polypeptide linker as an alternative to the corresponding f(ab')2 fragment. heavy- and light-chain genes of antibody-producing mouse hybridoma cell l ...19979085128
conventional and saturation-transfer epr of spin-labeled mutant bacteriophage m13 coat protein in phospholipid bilayers.a mutant of bacteriophage m13 was prepared in which a cysteine residue was introduced at position 25 of the major coat protein. the mutant coat protein was spin-labeled with a nitroxide derivative of maleimide and incorporated at different lipid-to-protein (l/p) ratios in dopc or dopg. the rotational dynamics of the reconstituted mutant coat protein was studied using epr and saturation transfer (st) epr techniques. the spectra are indicative for an anisotropic motion of the maleimide spin label ...19979247162
mutations that render the promoter of the histidine operon of salmonella typhimurium insensitive to nutrient-rich medium repression and amino acid downshift.the effects of mutations in the promoter of the histidine operon of salmonella typhimurium were examined in vivo. the wild-type chromosomal copy of the his promoter was replaced with mutations in the -10 hexamer sequence and in the region between the -10 hexamer and the transcriptional start point-termed the discriminator sequence. the substitutions were performed with a phage m13 allele replacement system. expression of the his operon is known to correlate with levels of guanosine 5',3'-bispyro ...19979260966
replication of m13 single-stranded dna bearing a site-specific ethenocytosine lesion by escherichia coil cell extracts.previous investigation on the mutagenic effects of 3, n4-ethenocytosine (epsilon c), a nonpairing dna lesion, revealed the existence of a novel sos-independent inducible mutagenic mechanism in e. coli termed uvm for uv modulation of mutagenesis. to investigate whether uvm is mediated by an alteration of dna replication, we have set up an in vitro replication system in which phage m13 viral single-stranded dna bearing a single site-specific (epsilon c) residue is replicated by soluble protein ext ...19979261557
in situ aggregational state of m13 bacteriophage major coat protein in sodium cholate and lipid bilayers.the in situ aggregational behavior of the bacteriophage m13 major coat protein was determined for the protein isolated in sodium cholate and reconstituted into dopc lipid bilayers. for this purpose, the cysteine mutants a49c and t36c of the major coat protein were labeled with either a maleimido spin-label or a fluorescence label (iaedans). the steric restrictions sensed by the spin-label were used to evaluate the local protein conformation and the extent of protein-protein interactions at the p ...19979315865
specificity of mutagenesis by 4-aminobiphenyl: mutations at g residues in bacteriophage m13 dna and g-->c transversions at a unique dg(8-abp) lesion in single-stranded dna.mutagenesis by the human bladder carcinogen 4-aminobiphenyl (abp) was studied in single-stranded dna from a bacteriophage m13 cloning vector. in comparison to abp lesions in double-stranded dna, lesions in single-stranded dna were approximately 70-fold more mutagenic and 50-fold more genotoxic. sequencing analysis of abp-induced mutations in the lacz gene revealed exclusively base-pair substitutions, with over 80% of the mutations occurring at g sites; the g at position 6310 accounted for 25% of ...19979450488
molecular cloning, expression, and characterization of a functional single-chain fv antibody to the mycotoxin zearalenone.the heavy-chain and kappa light-chain variable region genes of an antizearalenone hybridoma cell line (2g3-6e3-2e2) were isolated by pcr and joined by a dna linker encoding peptide (gly4ser)3 as a single-chain fv (scfv) dna fragment. the scfv dna fragment was cloned into a phagemid (pcantab5e) and expressed as a fusion protein with e tag and phage m13 p3 in escherichia coli tg1. in the presence of helper phage m13k07, the scfv fusion protein was displayed on the surfaces of recombinant phages. h ...19978979354
screening panels of monoclonal antibodies using phage-displayed antigen.a procedure is described to screen panels of hybridomas or purified monoclonal antibodies using antigen displayed on the surface of filamentous bacteriophage. in this system, samples containing murine monoclonal antibodies are incubated with phage-displayed antigen in microtiter plates coated with rabbit anti-mouse igg, and bound antibody-phage complex is detected with horseradish peroxidase-sheep anti-phage m13 conjugate. the assay has been validated with a panel of 16 monoclonal antibodies dir ...19979177746
identification of pathogenic yeasts of the imperfect genus candida by polymerase chain reaction fingerprinting.with the increase in the number of immunocompromised hosts, the number of fungal pathogens has increased markedly. identification and classification, especially of yeast species and strains, is often difficult when based solely on phenotypic characteristics. since it became clear that different fungal pathogens require specific treatment strategies, there is a need for simple, rapid and reliable methods to identify fungal isolates. polymerase chain reaction (pcr) fingerprinting was successfully ...19979378120
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 ...19989657049
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 ...19989482698
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 ...19989477227
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 ...19989665724
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) ...199812174274
purification and properties of a dna primase from nicotiana tabacum.a dna primase was isolated from a nuclear fraction from leaves of tobacco (nicotiana tabacum l. cv. samsun) and from purified nuclei prepared from tobacco suspension culture cells. the dna primase was purified to homogeneity (i) for preparations from leaves, by ammonium sulphate fractionation, followed by chromatography on columns of phosphocellulose, q-sepharose, heparin-sepharose and single-stranded dna cellulose, and sedimentation in a glycerol gradient, or (ii) for preparations from cells, b ...19989443386
a novel soluble tissue factor variant with an altered factor viia binding interface.tissue factor (tf) residues lys20 and asp58 form part of a binding epitope previously shown by alanine scanning to be critical for high affinity interactions with factor viia (fviia). to explore the possibility of enhancing the affinity of a tf-based antagonist for fviia, we created libraries in which residues at 20, 58, and adjacent positions were varied in constructs containing the soluble extracellular domain of tf (stf) fused to the bacteriophage m13 tail coat protein. tf variants monovalent ...19989461610
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 ...19989843943
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 ...19989827917
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.19989862799
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 ...19989672201
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 ...19989824155
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 ...19989705339
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 ...19989733645
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 ...19989735296
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 ...19989819049
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 ...19989531678
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 ...19989537524
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 ...19989557711
[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 ...199910330618
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 ...19999858539
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 ...199910381082
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 ...199910447883
[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 ...199910505264
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 ...199910536430
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 ...199910579091
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 ...199910668132
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 ...19999882367
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 ...199910219262
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 ...199910337489
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 ...199910329170
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 ...199910529252
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 ...19999894010
[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 ...199910420275
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 ...199910420969
[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 ...199910519075
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.200010714523
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.200010746750
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 ...200010858361
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 ...200010931196
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 ...200010945452
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 ...200010634904
rapid titration of multiple samples of filamentous bacteriophage (m13) on nitrocellulose filters. 200011126120
[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.200011798521
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 ...200010747951
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 ...200011118542
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 ...200011123944
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 ...200111708490
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 ...200111286974
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 ...200111307592
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 ...200111342716
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 ...200111286965
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 ...200111361332
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 ...200111472535
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 ...200111591151
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 ...200111755204
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, ...200111241275
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 ...200111255109
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 ...200111498772
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 ...200111549886
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 ...200111395866
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 ...200111397457
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 ...200111470533
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 ...200111346267
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 ...200111125152
ultrasensitive quartz crystal microbalance sensors for detection of m13-phages in liquids.quartz crystal microbalance (qcm) sensors are widely used for determining liquid properties or probing interfacial processes. for some applications the sensitivity of the qcm sensors typically used (5-20 mhz) is limited compared with other biosensor methods. in this study ultrasensitive qcm sensors with resonant frequencies from 39 to 110 mhz for measurements in the liquid phase are presented. the fundamental sensor effect of a qcm is the decrease of the resonant frequency of an oscillating quar ...200111679251
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 ...200211853411
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 ...200211690690
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 ...200212186783
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 ...200212234527
[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 ...200212018169
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 ...200212020787
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 ...200212021868
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 > ...200212044874
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 ...200211825608
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 ...200211825896
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 ...200312667684
mapping protein-protein interactions with phage-displayed combinatorial peptide libraries.this unit describes the process and analysis of affinity selecting bacteriophage m13 from libraries displaying combinatorial peptides fused to either a minor or major capsid protein. direct affinity selection uses target protein bound to a microtiter plate followed by purification of selected phage by elisa. alternatively, there is a bead-based affinity selection method. these methods allow one to readily isolate peptide ligands that bind to a protein target of interest and use the consensus seq ...200318228422
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 ...200312606054
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 ...200312609538
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 ...200312659940
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 ...200312660153
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- ...200312706896
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 ...200312938039
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.200312969496
[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 ...200312669420
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 ...200312738371
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 ...200312906514
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 ...200312875850
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