| site-directed mutagenesis of the m13 gene 5 protein: the role of arg-21, tyr-26 and tyr-41. | the gene 5 protein of bacteriophage m13 is a single stranded dna binding protein essential for phage replication. we have generated the mutations r21a, y26f and y41a in the gene 5 protein and purified the mutant proteins for functional characterisation in vitro. the complex of y26f with single-stranded dna is disrupted at 0.8 m nacl, the same salt concentration as that required to dissociate the native complex. however, the mutant proteins r21a and y41a are considerably less stable and dissociat ... | 1995 | 7999799 |
| m13 dna fingerprinting in differentiation of marine vibrio species. | the identification and differentiation of bacterial strains and species are frequently carried out by the use of diagnostic biochemical profiles, serology, and the detection of restriction fragment length polymorphisms in genomic dna. we show here that dna restriction fragment length polymorphisms detected using a probe derived from bacteriophage m13 can discriminate between several marine vibrio species. we have also demonstrated that individual isolates of vibrio species can be differentiated ... | 1994 | 8000478 |
| construction and characterization of m13 bacteriophages displaying gp120 binding domains of human cd4. | the envelope glycoprotein, gp120, on the surface of hiv interacts with the human cd4 molecule and thus helps the virus in gaining entry into the t-helper cells. to display the gp120 binding domains of human cd4 on the surface of the bacteriophage m13, two types of vectors have been constructed. in these, the first 176 amino acids of the human cd4 have been fused with the minor coat protein, giiip, of m13 bacteriophage for surface display. the western blot analysis revealed that using the phage b ... | 1994 | 8002012 |
| cleavable signal peptides are rarely found in bacterial cytoplasmic membrane proteins (review). | most proteins destined for secretion are synthesized with amino-terminal extensions, known as signal peptides, which play a vital role in their translocation across the membrane bordering the cytoplasm. following translocation across the eukaryotic endoplasmic reticulum (er) membrane or the bacterial cytoplasmic membrane, signal peptides are proteolytically removed from the preproteins. the process of membrane protein assembly can be likened to that of protein export in that it involves the tran ... | 1994 | 8019598 |
| a model of the complex between single-stranded dna and the single-stranded dna binding protein encoded by gene v of filamentous bacteriophage m13. | a contact analysis and a series of restrained molecular dynamics simulations were employed to derive a model of the complex between single-stranded dna and the single-stranded dna-binding protein encoded by gene v of the filamentous phage m13. the study is based on the recently elucidated solution structure of the tyr41-->his mutant of the protein. electron microscopy studies, indicating that the complex forms a flexible, left-handed helical coil with a diameter of 8 to 9 nm and an average pitch ... | 1994 | 8035458 |
| analysis of 31p mas nmr spectra and transversal relaxation of bacteriophage m13 and tobacco mosaic virus. | phosphorus magic angle spinning nuclear magnetic resonance (nmr) spectra and transversal relaxation of m13 and tmv are analyzed by use of a model, which includes both local backbone motions of the encapsulated nucleic acid molecules and overall rotational diffusion of the rod-shaped virions about their length axis. backbone motions influence the sideband intensities by causing a fast restricted reorientation of the phosphodiesters. to evaluate their influence on the observed sideband patterns, w ... | 1994 | 8038391 |
| high-affinity urokinase receptor antagonists identified with bacteriophage peptide display. | affinity selection of a 15-mer random peptide library displayed on bacteriophage m13 has been used to identify potent ligands for the human urokinase receptor, a key mediator of tumor cell invasion. a family of receptor binding bacteriophage ligands was obtained by sequentially and alternately selecting the peptide library on cos-7 monkey kidney cells and baculovirus-infected sf9 insect cells overexpressing the human urokinase receptor. nineteen peptides encoded by the random dna regions of the ... | 1994 | 8041758 |
| structure of a foreign peptide displayed on the surface of bacteriophage m13. | the use of filamentous bacteriophage m13 as a vehicle for display of foreign peptides and proteins provides a means for the construction of therapeutic, diagnostic and technological tools of broad utility. the usefulness of this technology is dependent on the ability of an inserted peptide to act as a ligand when fused to a structural protein. this, in turn, depends on the configuration in which the fused peptide is presented on the surface of the phage. x-ray diffraction from oriented fibers of ... | 1994 | 8057360 |
| recognition by human sera and immunogenicity of hbsag mimotopes selected from an m13 phage display library. | we used two mouse monoclonal antibodies (mab) specific for the human hepatitis b virus surface antigen (hbsag) to screen a random peptide library of 15 amino-acid residues displayed as a fusion to protein iii of filamentous phage m13. by a combination of affinity selection, immuno-screening and elisa techniques, we selected peptides that are recognized by the anti-hbsag mab and show aa similarity with the natural antigen. the selected phage-displayed epitopes (phagotopes) behave as antigenic mim ... | 1994 | 8076818 |
| quencher-enhanced specificity of psoralen-photosensitized virus inactivation in platelet concentrates. | treatment with psoralens and uva (puva) has been shown to be efficacious in eliminating the risk of virus transmission by platelet concentrates (pcs). it has previously been demonstrated that, during the inactivation of cell-free vesicular stomatitis virus (vsv) by aminomethyltrimethylpsoralen (amt) and uva in pcs, platelet function could be protected either by oxygen removal before irradiation or by inclusion of a type i free radical quencher, such as mannitol. | 1994 | 8091471 |
| selection of an anti-igf-1 fab from a fab phage library created by mutagenesis of multiple cdr loops. | diverse fab libraries containing 2-3 x 10(8) members were generated by randomizing amino acid residues within four of the six complementarity determining regions of a humanized version of an anti-her-2 ab (hu4d5). these libraries were subsequently displayed on the surface of the filamentous bacteriophage m13 and selected for binding to three proteins: cd4, insulin-like growth factor 1 (igf-1), and tissue plasminogen activator. an fab-bacteriophage was isolated that showed specific binding to igf ... | 1993 | 8099557 |
| kinetic parameters of the translocation of bacteriophage t4 gene 41 protein helicase on single-stranded dna. | the bacteriophage t4 gene 41 helicase protein (gp41) carries a single-stranded dna-dependent atpase activity that is essential to its helicase activity. this atpase activity can be stimulated by a wide variety of single-stranded dna cofactors, including homo-oligomers and homopolymers 8 to approximately 10,000 nucleotide residues in length, and by natural single-stranded dna, such as bacteriophage m13 dna. the steady-state atpase activity of gp41 on single-stranded homopolymeric cofactors is dep ... | 1994 | 8107085 |
| the solution structure of the tyr41-->his mutant of the single-stranded dna binding protein encoded by gene v of the filamentous bacteriophage m13. | the solution structure of mutant tyr41-->his of the single-stranded dna binding protein encoded by gene v of the filamentous bacteriophage m13 has been investigated by nuclear magnetic resonance spectroscopy. two- and three-dimensional nmr experiments have been employed with a variety of nmr samples of gene v protein, some of which were uniformly enriched with either 15n or 13c. the structure of mutant tyr41-->his of the m13 gene v protein which occurs in solution as a symmetric dimer was calcul ... | 1994 | 8107108 |
| ingested foreign (phage m13) dna survives transiently in the gastrointestinal tract and enters the bloodstream of mice. | is the epithelial lining of the mammalian gastrointestinal (gi) tract a tight barrier against the uptake of ingested foreign dna or can such foreign dna penetrate into the organism? we approached this question by pipette-feeding circular or linearized double-stranded phage m13 dna to mice or by adding m13 dna to the food of mice whose fecal excretions had previously been shown to be devoid of this dna. at various post-prandial times, the feces of the animals was tested for m13 dna sequences by s ... | 1994 | 8121408 |
| differentiation of bacillus anthracis from other bacillus cereus group bacteria with the pcr. | variation among isolates of bacillus anthracis was examined by using restriction fragmentation patterns and the pcr performed with arbitrary and sequence-specific oligonucleotide primers. the patterns were compared with the patterns generated from strains of closely related species belonging to the "bacillus cereus group" of bacteria, including b. cereus, bacillus thuringiensis, and bacillus mycoides. all b. anthracis profiles were identical for each of 18 restriction enzymes, each of 10 arbitra ... | 1994 | 8123566 |
| a direct selection strategy for shotgun cloning and sequencing in the bacteriophage m13. | a new cloning strategy is described which utilizes direct selection of recombinants for shotgun sequencing in the filamentous bacteriophage m13. direct selection is accomplished by insertional inactivation of the m13 gene x protein, a powerful inhibitor of phage-specific dna synthesis when overproduced. an extra copy of gene x was inserted into the intergenic region of m13 and placed under the control of the bacteriophage t7 gene 10 promoter and rbs. random fragments are cloned into the ecorv cl ... | 1994 | 8127645 |
| identification of three n-linked glycans in the v4-v5 region of hiv-1 gp 120, dispensable for cd4-binding and fusion activity of gp 120. | site-directed mutagenesis was used to study the biological significance of three n-linked glycans (linked to asn406, asn448, and asn463), situated in the cd4-binding region of gp120. mutagenesis was carried out in a phage m13 system, and the mutated env genes were inserted into recombinant vaccinia virus (r-vaccinia virus). to evaluate if the level of expression affected the biological phenotype of mutant gp120, we expressed the envelope glycoproteins using either a weak (7.5 k) or a strong (11 ... | 1994 | 8129620 |
| isolation of neutralizing anti-c5a monoclonal antibodies from a filamentous phage monovalent fab display library. | a panel of mabs against the activated complement component c5a was obtained from a filamentous phage m13-fab display library generated from mice immunized with human rc5a. fabs isolated from the library after iterative selection vs rc5a bound to both rc5a and purified c5. to isolate fabs specific for neoepitopes expressed on c5a but not on the native complement component c5 the library was rescreened in a competitive manner. the phage fab library was first incubated with immobilized c5 to deplet ... | 1994 | 8157971 |
| a novel intergenic site for integration and expression of foreign genes in the genome of pseudorabies virus. | restriction enzyme analysis of dna of a number of pseudorabies virus (prv) single plaque isolates revealed in several cases the existence of a unique ecori cleavage site, which has not been observed in prv dna before. this ecori site was mapped to the right end of the unique long region of the prv genome, in bamhi-fragment 6. sequence analysis of this region demonstrated the presence of an 11 bp tandem repeat in variable copy numbers in different prv strains, suggesting the creation of the ecori ... | 1994 | 8175950 |
| sequence analysis of the lysin gene region of the prolate lactococcal bacteriophage c2. | approximately 80% of the genome of the prolate-headed lactococcal bacteriophage c2 was cloned into shuttle vectors psa3 and pfx3 in escherichia coli and transferred to lactococcus lactis. a 1.67-kilobase ecorv fragment containing the gene for the phage lysin was identified and the position and orientation of the phage lysin gene in the physical map of the phage were determined. the phage lysin was expressed in e. coli and its sequence was determined and compared with the sequences of other bacte ... | 1993 | 8221377 |
| mutagenic and genotoxic effects of three vinyl chloride-induced dna lesions: 1,n6-ethenoadenine, 3,n4-ethenocytosine, and 4-amino-5-(imidazol-2-yl)imidazole. | the mutagenic and genotoxic properties of 1,n6-ethenoadenine (epsilon ade), 3,n4-ethenocytosine (epsilon cyt), and 4-amino-5-(imidazol-2-yl)imidazole (beta) were investigated in vivo. the former two modified bases are known dna adducts formed by the human carcinogen vinyl chloride; beta is formed by pyrimidine ring-opening of epsilon ade. chemically synthesized deoxyhexanucleotides containing epsilon ade and beta, d[gct-(epsilon a)gc], and d[gct(beta)gc], respectively, were described previously ... | 1993 | 8251500 |
| the dna sequence of adenovirus type 40. | the 34,214 bp dna sequence of adenovirus type 40 strain dugan was determined directly from random fragments of virion dna cloned into a bacteriophage m13 cloning vector. the gene layout is similar to that of other human adenoviruses, and in addition contains two potential protein-coding regions that are conserved, but have not been recognized previously, in other adenovirus genomes. one is oriented rightward, contained within the intron in the protein-coding region for the l4 33k gene, and would ... | 1993 | 8263936 |
| val-->ala mutations selectively alter helix-helix packing in the transmembrane segment of phage m13 coat protein. | val-->ala mutations within the effective transmembrane segment of a model single-spanning membrane protein, the 50-residue major coat (gene viii) protein of bacteriophage m13, are shown to have sequence-dependent impacts on stabilization of membrane-embedded helical dimeric structures. randomized mutagenesis performed on the coat protein hydrophobic segment 21-39 (yigyawamv-vvivgatigi) produced a library of viable mutants which included those in which each of the four valine residues was replace ... | 1993 | 8265602 |
| rapid identification and differentiation of yeasts by dna and pcr fingerprinting. | we have used the techniques of dna fingerprinting and polymerase chain reaction (pcr) with probes specific for hypervariable repetitive dna sequences (mini- and microsatellite dnas) to analyze 36 yeast strains belonging to 10 species and 2 genera. using (gtg)5, (gaca)4, phage m13 dna and the m13 sequence gagggtggcggttct as probes and primers, respectively, we obtained dna polymorphisms which allowed us to discriminate 23 biotechnologically important strains of the yeast saccaromyces cerevisiae a ... | 1993 | 8271158 |
| [analysis of peptide fragment insertions in the basic proteins of the bacteriophage m13, f1, and fd envelopes. interconnection of structural characteristics of the envelope protein and viability of mutant phages]. | an analysis of 12 peptide fragment insertions into the major coat protein (protein pviii) of bacteriophages m13, f1 and fd has been done. to elucidate the relations between protein structural characteristics and viability of mutant phages, we used the program pro-anal. correlations were found between phage viability and different physicochemical and structural characteristics of protein n-termini. thus peptide insertions in nonviable phages have high indexes of alpha-helicity, volumes, and polar ... | 1993 | 8283982 |
| [family analysis of human "fingerprints" obtained using a phage m13 dna probe]. | the human "fingerprints" detected by the phage m13 dna probe from 40 simple families with some presumably genetic malformations were observed. when only one parent had a specific band, the mean segregation frequency of all the electrophoretic bands detected was equal to 0.485, which confirmed the hypothesis about the mendelian inheritance and high level of heterozygosity of hypervariable loci. the mean new measure of similarity between the parents calculated by li c.c. was 0.54 (this measure wit ... | 1993 | 8307351 |
| [genetic distances between various ethnic populations calculated on the basis of polymorphism of dna detected by the hypervariable phage m13 dna probe]. | the frequencies of different electrophoretic bands in dna "fingerprint" detected by the phage m13 dna probe in six russian populations from the kirov district and krasnodar, one from chuvashia and one from tuva were compared with each other and pseudo-genetic distances by nei were calculated. the results corresponded well with a presumable extent of similarity between the populations observed. | 1993 | 8307352 |
| identification of a domain of escherichia coli primase required for functional interaction with the dnab helicase at the replication fork. | primase plays a key role in governing the sequence of events required on the lagging strand during a cycle of okazaki fragment synthesis. to begin to probe the protein-protein interactions necessary for primase function at the replication fork, we have used limited trypsinolysis to separate primase into two functional domains, an n-terminal domain of 49 kda (p49) and a carboxyl-terminal domain of 16 kda (p16). p49 retained primase activity in replication assays that utilized bacteriophage m13 dn ... | 1994 | 8308039 |
| nucleosome assembly during complementary dna strand synthesis in extracts from mammalian cells. | circular single-stranded phage m13 dna is used as a template for complementary strand synthesis in cytosolic extracts from proliferating hela cells. dna synthesis is initiated by one or maximally two priming events and typically leads to covalently closed double-stranded reaction products. when carried out in the presence of the nuclear chromatin assembly factor caf-1, complementary strand synthesis is accompanied by nucleosome assembly. this novel system is very useful for the study of basic bi ... | 1993 | 8314801 |
| assignment of 1h, 15n, and backbone 13c resonances in detergent-solubilized m13 coat protein via multinuclear multidimensional nmr: a model for the coat protein monomer. | the major coat protein (gviiip) of bacteriophage m13 complexed with sds detergent micelles was used as a model system to study the lipid-bound conformation of the protein. conditions were found that allowed the recording of good quality of nmr spectra. by making extensive use of three-dimensional heteronuclear (13c, 15n) nmr, we obtained a complete set of resonance assignments for 1hn, 1h alpha, 1h beta, 13c alpha, co, and 15n and partially assigned the rest of the 1h spectrum. analysis of noe a ... | 1993 | 8347628 |
| [application of dna fingerprinting to investigation of genetic relationships between laboratory rabbit strains]. | it is well known that laboratory rabbits are not controlled genetically like laboratory mice and rats. in order to test the usefulness of dna fingerprinting in investigation of genetic uniformity of the laboratory rabbits strains and their relationships, we applied dna fingerprinting using bacteriophage m13 probe to five strains (2 inbreds (jwy-nibs and duy-nibs) and 3 outbreds (jw-nibs, icl:jw and whhl)). dna fingerprints of 2 inbred strains showed the same banding patterns within each strain b ... | 1993 | 8354354 |
| receptor phage. display of functional domains of the human high affinity ige receptor on the m13 phage surface. | in this paper we demonstrate that phage display technology is a suitable system for studying the interaction between the high-affinity receptor for ige (fc epsilon ri) and ige. the alpha subunit extracellular domains of the human receptor were expressed on the surface of filamentous phage m13 fused to the carboxyl-terminal part of the gene iii protein (piii). two constructs were made, the first with both the ig-like domains of the receptor alpha chain and the second with only the c-terminal doma ... | 1993 | 8354400 |
| a theoretical study of rotational diffusion models for rod-shaped viruses. the influence of motion on 31p nuclear magnetic resonance lineshapes and transversal relaxation. | information about the interaction between nucleic acids and coat proteins in intact virus particles may be obtained by studying the restricted backbone dynamics of the incapsulated nucleic acids using 31p nuclear magnetic resonance (nmr) spectroscopy. in this article, simulations are carried out to investigate how reorientation of a rod-shaped virus particle as a whole and isolated nucleic acid motions within the virion influence the 31p nmr lineshape and transversal relaxation dominated by the ... | 1993 | 8369411 |
| analysis of 31p nuclear magnetic resonance lineshapes and transversal relaxation of bacteriophage m13 and tobacco mosaic virus. | the experimentally observed 31p lineshapes and transversal relaxation of 15% (wt/wt) m13, 30% m13, and 30% tobacco mosaic virus (tmv) are compared with lineshapes and relaxation curves that are simulated for various types of rotational diffusion using the models discussed previously (magusin, p. c. m. m., and m. a. hemminga. 1993. biophys. j. 64:1851-1860). it is found that isotropic diffusion cannot explain the observed lineshape effects. a rigid rod diffusion model is only successful in descri ... | 1993 | 8369412 |
| exploration of the single-stranded dna-binding domains of the gene v proteins encoded by the filamentous bacteriophages ike and m13 by means of spin-labeled oligonucleotide and lanthanide-chelate complexes. | scrutiny of noe data available for the protein encoded by gene v of the filamentous phage ike (ike gvp), resulted in the elucidation of a beta-sheet structure which is partly five stranded. the dna-binding domain of ike gvp was investigated using a spin-labeled deoxytrinucleotide. the paramagnetic-relaxation effects observed in the 1h-nmr spectrum of ike gvp, upon binding of this dna fragment, could be visualized using two-dimensional difference spectroscopy. in this way, the residues present in ... | 1993 | 8375389 |
| m13 and puc vectors with new unique restriction sites for cloning. | several vectors based on the widely used phage m13 and plasmid puc were constructed. the vectors contain polylinkers (mcs) for dna insertions with several new restriction sites (e.g., apai, noti, stui, sacii). moreover, the nari site in the nonessential part of the vector molecule was changed into a bsshii site, so that the nari site in the mcs became unique. | 1993 | 8393824 |
| exploring the dna binding domain of gene v protein encoded by bacteriophage m13 with the aid of spin-labeled oligonucleotides in combination with 1h-nmr. | the dna binding domain of the single-stranded dna binding protein gene v protein encoded by the bacteriophage m13 was studied by means of 1h nuclear magnetic resonance, through use of a spin-labeled deoxytrinucleotide. the paramagnetic relaxation effects observed in the 1h-nmr spectrum of m13 gvp upon binding of the spin-labeled ligand were made manifest by means of 2d difference spectroscopy. in this way, a vast data reduction was accomplished which enabled us to check and extend the analysis o ... | 1993 | 8396429 |
| the macroscopic organization of reconstituted m13 coat protein-phospholipid systems. an epr spectroscopy and polarizing microscope study. | the coat protein of the bacteriophage m13 in the alpha-helical state is reconstituted in macroscopically oriented systems of dioleoylphosphatidylcholine that are prepared by squeezing the reconstituted material between glass plates. the coat protein dramatically influences the macroscopic orientation of the multibilayers, as is investigated by polarizing microscopy and epr spectroscopy of the cholestane spin label embedded in the bilayers. it is found that with increasing amounts of protein the ... | 1993 | 8399296 |
| dna- and pcr-fingerprinting in fungi. | dna-fingerprinting has been successfully used to detect hypervariable, repetitive dna sequences (minisatellites and microsatellites) in fungi. combined with methods used to identify random amplified polymorphic dna (rapd), conventional dna-fingerprinting hybridization probes can also be used as single primers to detect dna polymorphisms among fungal species and strains. the oligonucleotides (ca)8, (ct)8, (cac)5, (gtg)5, (gaca)4 and (gata)4, as well as the phage m13 and its core sequence, have be ... | 1993 | 8400701 |
| hybridization probes for conventional dna fingerprinting used as single primers in the polymerase chain reaction to distinguish strains of cryptococcus neoformans. | in conventional dna fingerprinting, hypervariable and repetitive sequences (minisatellite or microsatellite dna) are detected with hybridization probes. as demonstrated here, these probes can be used as single primers in the polymerase chain reaction (pcr) to generate individual fingerprints. several conventional dna fingerprinting probes were used to prime the pcr, yielding distinctive, hypervariable multifragment profiles for different strains of cryptococcus neoformans. pcr fingerprinting wit ... | 1993 | 8408543 |
| mutagenicity and genotoxicity of the major dna adduct of the antitumor drug cis-diamminedichloroplatinum(ii). | the mutagenicity and genotoxicity of cis-[pt(nh3)2[d(gpg)-n7(1),-n7(2)]] (g*g*), the major dna adduct of the antitumor drug cisplatin, has been investigated in escherichia coli. a duplex bacteriophage m13 genome was constructed to contain the g*g* adduct at a specific site in the (-) strand. the singly platinated duplex genome exhibited a survival of 22% relative to that of the unplatinated control genomes, and this value rose to 38% in cells treated with ultraviolet light to induce the sos resp ... | 1993 | 8422401 |
| conformational states of mutant m13 coat proteins are regulated by transmembrane residues. | mutational and structural analysis of the 28 viable bacteriophage m13 mutants obtained by randomized mutagenesis of the effective transmembrane (tm) segment of the 50-residue major coat (gene viii) protein (residues 21-39) demonstrated that m13 coat protein functionality, as reflected by phage viability, is incompatible with an increase in gly + beta-branched residue content in its tm core. sds-polyacrylamide gel electrophoresis and circular dichroism spectroscopy performed in membrane environme ... | 1993 | 8444834 |
| cloning, expression and in vitro characterisation of the m13 gene 5 protein. | the gene 5 protein encoded in the genome of bacteriophage m13 is a single stranded dna binding protein essential for phage replication. we have cloned a fragment of the m13 genome containing gene 5, and investigated the effect of upstream elements on expression of the gene by means of bal 31 deletion analysis. the gene was also expressed from the lac promoter of the phagemid vector puc119, and the recombinant protein purified and characterised for dna binding. the affinity of the recombinant pro ... | 1993 | 8504168 |
| phage display as a rapid gene expression system: production of bioactive cytokine-phage and generation of neutralizing monoclonal antibodies. | proteins, such as hormones, enzymes, or antibody binding sites, can be expressed in an active conformation on the surface of filamentous bacteriophage. although the phage display technology was originally developed for binding studies, we demonstrate here that this technique can rapidly provide cytokines for studies of biological activity and for raising neutralizing monoclonal antibodies. a phage m13-based cloning vector was constructed that facilitated the expression of human interleukin 3 (hi ... | 1993 | 8505547 |
| trypsin display on the surface of bacteriophage. | the gene iii and viii-encoded coat proteins (piii and pviii) from bacteriophage m13 have been fused to the c terminus of the serine protease, trypsin (tsn). the genes encoding the fusions were then inserted directly into m13mp18 to create vectors which expressed both the tsn-coat protein hybrids and the wild-type (wt) coat proteins. immunoblot analysis confirmed that the bacteriophage express tsn on their surface. isolated fusion phage possess kinetic parameters which approximate those of the wt ... | 1993 | 8508953 |
| [dna fingerprinting in horses]. | using a multilocus dna probe, individual - specific hybridization patterns, the so-called dna fingerprints (tab) were determined in six horse families by the dna fingerprinting method. the probe with evolutionally preserved nucleotide sequence from bacteriophage m13 determines hypervariable regions placed in genomic minisatellite dna. the use of this probe permits an identification of an individual and execution of paternity relationships with a probability over 99.99 per cent. | 1993 | 8511839 |
| pcr-fingerprinting used for comparison of ex type strains of trichoderma species deposited in different culture collections. | pcr-fingerprinting with primers (gaca)4, (gtg)5, m13 (core sequence of phage m13), and opb-05 was used to compare ex type strains of various species of trichoderma. the ex type strain of each species was obtained from different culture collections. pcr-fingerprints of each ex type culture, despite of separate cultivation over a long period of time (sometimes decades) by different culture collections, were identical. ex type strains could be discriminated from a number of other strains belonging ... | 1995 | 8564364 |
| molecular characterization and cytogenetic analysis of highly repeated dnas of lake trout, salvelinus namaycush. | the chromosomes of lake trout (salvelinus namaycush) contain a considerable amount of heterochromatin located at the centromeres and/or telomeres of several chromosomes, including a sex-specific block located distally on the x chromosome. in order to investigate further the repetitive dnas of lake trout, genomic dna from a female was size fractionated (<600 bp) with the restriction endonuclease alui and fragments were cloned into the bacteriophage m13. a total of 42 clones were isolated. relativ ... | 1995 | 8565700 |
| polymerase chain reaction fingerprinting in fungi using single primers specific to minisatellites and simple repetitive dna sequences: strain variation in cryptococcus neoformans. | minisatellites and simple repetitive dna sequence motifs are used as conventional oligonucleotide probes in dna-hybridization-based fingerprinting. the same oligonucleotides can be used as single primers in the polymerase chain reaction (pcr) to generate individual pcr fingerprints. in this study, the simple repetitive sequences, (ca)8, (ct)8, (cac)5, (gtg)5, (gaca)4 and (gata)4, and a minisatellite core sequence derived from the wild-type phage m13 (5' gagggtggcggttct 3') were used as specific, ... | 1995 | 8582350 |
| protein-protein interactions between the escherichia coli single-stranded dna-binding protein and exonuclease i. | it was demonstrated previously that a deoxyribophosphodiesterase (drpase) activity is associated with the dna repair enzyme exonuclease i, and that this activity is stimulated by the addition of the e. coli single-stranded dna-binding protein (ssb). this activity catalyzes the release of deoxyribose-phosphate groups at apurinic/apyrimidinic (ap) sites in the dna that have been cleared by the action of an ap endonuclease. we have now used the yeast two-hybrid system to demonstrate that a protein- ... | 1996 | 8619028 |
| mutational properties of the primary aflatoxin b1-dna adduct. | the mutagenic activity of the major dna adduct formed by the liver carcinogen aflatoxin b1 (afb1) was investigated in vivo. an oligonucleotide containing a single 8,9-dihydro-8-(n7-guanyl)-9-hydroxyaflatoxin b1 (afb1-n7-gua) adduct was inserted into the single-stranded genome of bacteriophage m13. replication in sos-induced escherichia coli yielded a mutation frequency for afb1-n7-gua of 4%. the predominant mutation was g --> t, identical to the principal mutation in human liver tumors believed ... | 1996 | 8643667 |
| cloning aspergillus fumigatus allergens by the pjufo filamentous phage display system. | a cdna library from aspergillus fumigatus has been displayed on the surface of filamentous phage m13 and screened for gene products binding to human serum ige. the physical linkage of cdna gene products to the genetic information required for their production, achieved by exploiting the high-affinity interaction of the jun and fos leucine zippers, allows rapid and easier screening of large libraries in semifluid systems. the pjufo cloning vector is designed to display proteins on the surface of ... | 1996 | 8645976 |
| lysogenic conversion by a filamentous phage encoding cholera toxin. | vibrio cholerae, the causative agent of cholera, requires two coordinately regulated factors for full virulence: cholera toxin (ct), a potent enterotoxin, and toxin-coregulated pili (tcp), surface organelles required for intestinal colonization. the structural genes for ct are shown here to be encoded by a filamentous bacteriophage (designated ctxphi), which is related to coliphage m13. the ctxphi genome chromosomally integrated or replicated as a plasmid. ctxphi used tcp as its receptor and inf ... | 1996 | 8658163 |
| reactions of mitochondrial cruciform cutting endonuclease 1 (cce1) of yeast saccharomyces cerevisiae with branched dnas in vitro. | cruciform-cutting endonuclease 1 (cce1) is an x-solvase from yeast saccharomyces cerevisiae [kleff, s., kemper, b. & sternglanz, r. (1992) embo j. 11, 699-704]. we report here the purification of the cloned enzyme cce1 to near homogeneity from over-expressing escherichia coli cells. the purified protein has a globular shape and an apparent molecular mass of 38 kda. cce1 reacts specifically with branched dnas, preferably with four-armed cruciforms. the enzyme linearizes native supercoiled dna by ... | 1996 | 8665955 |
| iterative optimization of high-affinity proteases inhibitors using phage display. 1. plasmin. | we generated a series of libraries having variants of the first kunitz domain of human lipoprotein-associated coagulation inhibitor (laci-d1, also known as tissue-factor pathway inhibitor-i) displayed on bacteriophage m13 as piii-fusions. we varied laci-di iteratively in two regions: the p1 region (positions 10-21) and the "second loop", (positions 31-39), which together form one end of the domain. display-phage library lib#1 allows 31 200 amino-acid sequences in p1 region (residues 13, 16-19). ... | 1996 | 8672509 |
| [detection of bovine infectious rhinotracheitis virus by hybridization using nonradioactive dna-probes]. | biotin-labeled dna probes for infectious bovine rhinotracheitis virus also known as bovine herpesvirus-1 (bhv-1) have been developed. the procedure is based on dot-blot hybridization using biotin-labeled bacteriophage m13 and plasmid probes containing cloned psti and ecori-psti restriction fragments of viral genome. the probes obtained were used to detect viral nucleic acids in specimens of bovine spermatic fluid or nasal swabs of calves. the method is simple and rapid, taking less than 24 h, an ... | 1995 | 8686268 |
| the solution structure of the synthetic circular peptide cgvsrqgkpyc. nmr studies of the folding of a synthetic model for the dna-binding loop of the ssdna-binding protein encoded by gene v of phage m13. | the cyclic disulfide peptide cgvsrqgkpyc was prepared to obtain a constrained analogue of residues 17-27 of the dna-binding loop of the gene-v-encoded sdna-binding protein of filamentous bacteriophage m13. amino acid sequences very similar to that of the beta-loop have been found in various phage-encoded ssdna-binding proteins, and it has been proposed that such a loop may occur as a common motif in this class of proteins. the conformation, in aqueous solution, of the synthetic gene-v-protein bi ... | 1996 | 8706671 |
| design of immunogens as components of a new generation of molecular vaccines. | three new approaches to design effective immunogens are considered. at first, we derived an expression vector from bacteriophage m13 allowing the exposure of short peptides on the virion surface. eia demonstrates that antibodies against a recombinant phage carrying the antigenic determinant of the hiv-1 gag protein reacted with the 17-kda core protein of the virus and also with its polyprotein precursor p55 in immunoblotting. in another approach, we chose the hepatitis b core antigen (hbcag) par ... | 1996 | 8717396 |
| concordance of clinical and environmental isolates of cryptococcus neoformans var. gattii by random amplification of polymorphic dna analysis and pcr fingerprinting. | sixty-one clinical and forty-nine environmental isolates of cryptococcus neoformans var. gattii from australia and the united states were analyzed by random amplification of polymorphic dna (rapd), using 12- to 22-mer primers in pairs, and/or pcr fingerprinting with a single primer derived from the microsatellite core sequence of the wild-type phage m13 (5' gagggtggcggttct 3'). three major genetic profiles were identified by both typing techniques. a single rapd profile (vgi) predominated among ... | 1996 | 8727912 |
| genetic dissimilarity of two fluconazole-resistant candida albicans strains causing meningitis and oral candidiasis in the same aids patient. | we describe a patient with aids who simultaneously developed candida meningitis with three positive cerebrospinal fluid cultures and oral candidiasis. this patient also had a history or recurrent episodes of oral candidiasis treated with fluconazole. the patient did not respond to this therapy but was cured with amphotericin b and flucytosine. in vitro susceptibility tests revealed that each infection was caused by fluconazole-resistant candida albicans isolates. strain delineation by karyotypin ... | 1996 | 8735114 |
| molecular typing by random amplification of polymorphic dna and m13 southern hybridization of related paired isolates of aspergillus fumigatus. | three forms of dna-based typing procedures for aspergillus fumigatus isolates have been developed over the last five years. the procedures are random amplification of polymorphic dna (rapd), restriction fragment length polymorphism (rflp) detection, and southern hybridizations with various repetitive sequence-based probes. using two of these procedures, we compared 16 selected isolates, grouped into eight pairs on the basis of epidemiology or previously assigned rflp types. rapd with four primer ... | 1996 | 8748280 |
| accessibility and environment probing using cysteine residues introduced along the putative transmembrane domain of the major coat protein of bacteriophage m13. | the major coat protein of the filamentous bacteriophage m13 is located in the inner membrane of host cell escherichia coli prior to assembly into virions. to identify the transmembrane domain of the coat protein, we have introduced unique cysteine residues along the putative transmembrane domain at position 25, 31, 33, 36, 38, 46, 47, 49, or 50. the mutant major coat protein was solubilized by membrane-mimicking detergents or reconstituted into mixed bilayers of phospholipids. information about ... | 1996 | 8756694 |
| [genomic polymorphism in mycobacterium tuberculosis strains]. | different genomic fingerprinting techniques (universal probes, such as rrna genes, phage m13 dna, is 6110 probe) have been used to investigate the genomic polymorphism of mycobacterium tuberculosis strains isolated in different geographical regions of russia and in some cis countries. as shown with the use of these techniques and a specially developed pcr-mediated system for genetic typing, m.tuberculosis strains are genotypically heterogeneous in regions with a sporadic level of tuberculosis mo ... | 1996 | 8771734 |
| anti-vaccinia virus effect of m13 bacteriophage dna. | single-stranded dna derived from m13 phage was evaluated for antiviral activity in mice infected with vaccinia virus. m13 dna at a dose as low as 16.7 mg/kg was effective in reducing the number of tail lesions caused by vaccinia virus by more than 90%. a single administration of m13 dna 1 day before infection was sufficient to reduce significantly the number of tail lesions caused by vaccinia virus. denatured eukaryotic nucleic acids such as calf thymus dna and human placenta dna were not effect ... | 1996 | 8793011 |
| partial cviji digestion as an alternative approach to generate cosmid sublibraries for large-scale sequencing projects. | we demonstrate an alternative method for the generation of random subclones for large-scale shotgun human dna sequencing projects. miniprep dna from a human cosmid clone was partially digested with cviji, size-fractionated by agarose gel electrophoresis and cloned into bacteriophage m13. a library consisting of 10(5) subclones of 1.1 kb average size was recovered from one gel fraction containing approximately 300 ng of partially digested dna. dna sequences from an initial 103 subclones demonstra ... | 1996 | 8816243 |
| a new concept in (adenoviral) oncogenesis: integration of foreign dna and its consequences. | a new concept for viral oncogenesis is presented which is based on experimental work on the chromosomal integration of adenovirus dna into mammalian genomes. the mechanism of adenovirus dna integration is akin to non-sequence-specific insertional recombination in which patch homologies between the recombination partners are frequently observed. this reaction has been imitated in a cell-free system by using nuclear extracts from hamster cells and partly purified fractions derived from them. as a ... | 1996 | 8876634 |
| analysis of peptide-binding motifs for two disease associated hla-dr13 alleles using an m13 phage display library. | major histocompatibility complex (mhc) molecules bind peptides bearing an appropriate 'sequence motif' for mhc binding. the use of phage display libraries exploits the ability of mhc class ii molecules to exchange peptides in solution and thus select out peptide sequences with high-affinity binding from a large array of random peptides. we have analysed the peptide binding motifs of hla-drb1*1301 and *1302 using affinity purified hla-dr13 molecules to purify sequentially hla-dr13-binding peptide ... | 1996 | 8881746 |
| analyzing sequencing reactions from bacteriophage m13 by matrix-assisted laser desorption/ionization mass spectrometry. | the current demand for improved dna sequencing methodologies posed by the human genome project has spurred the investigation of alternatives to gel electrophoresis. matrix-assisted laser desorption/ionization (maldi) mass spectrometry has great potential for the rapid analysis of dna fragments. mock sanger sequencing mixtures have been successfully analyzed by maldi by pooling synthesized oligonucleotides corresponding to the m13 bacteriophage sequence. more recently, analyses of sanger sequenci ... | 1996 | 8885418 |
| an improved method for the synthesis of mercurated dutp. enzymic synthesis of hg-labelled dna of high molecular weight suitable for use in an image based dna sequencing strategy. | the development of high-resolution scanning-probe microscopes has reawakened interest in the possibility of sequencing large nucleic acid molecules by direct imaging. such an approach would be facilitated by the availability of effective methods for increasing contrast by labelling specific nucleotides, and the utility of introducing mercury atoms into complete dna molecules through the enzymic polymerisation of mercurated pyrimidine deoxynucleoside triphosphates has been re-investigated. a simp ... | 1996 | 8912922 |
| stable high-copy-number bacteriophage lambda promoter vectors for overproduction of proteins in escherichia coli. | the construction of new high-copy-number (hcn) lambda-promoter expression vectors is described. all these vectors (1) contain tandem lambda pr and pl promoters upstream of an extensive multiple cloning site (mcs) for insertion of genes, (2) direct expression of the lambda cits857 gene, enabling their use in any escherichia coli host strain for thermal induction of gene overexpression, and (3) bear the par locus of plasmid psc101, ensuring their stable maintenance at hcn in the absence of continu ... | 1996 | 8918231 |
| [identification and classification of strains of microorganisms using genomic fingerprinting with biotinylated phage m13 dna]. | to analyze dna polymorphisms of various bacterial strains, a nonradioactive variant of the genomic fingerprinting method was developed. the method was based on the application of biotin-labeled single-stranded phage m13 dna as a probe. characteristic patterns of fingerprints obtained by mvai, haeiii, and hinfi restriction enzymes are presented for several species of bacilli and other bacteria. the advantages of this method in microbiology for the identification and characterization of different ... | 1996 | 8964460 |
| organization of the bacillus subtilis 168 chromosome between kdg and the attachment site of the sp beta prophage: use of long accurate pcr and yeast artificial chromosomes for sequencing. | within the bacillus subtilis genome sequencing project, the region between lysa and ilva was assigned to our laboratory. in this report we present the sequence of the last 36 kb of this region, between the kdg operon and the attachment site of the sp beta prophage. a two-step strategy was used for the sequencing. in the first step, total chromosomal dna was cloned in phage m13-based vectors and the clones carrying inserts from the target region were identified by hybridization with a cognate yea ... | 1996 | 8969496 |
| 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 ... | 1997 | 8979354 |
| destabilizing interactions between the partners of a bifunctional fusion protein. | hybrid male-gvp is a bifunctional protein in vitro since it binds maltose as protein male of escherichia coli and since it is dimeric and specifically binds single-stranded dna as protein gvp of phage m13. the oxidation rate of a unique cysteine residue was used to compare the stabilities of gvp in its free and hybrid forms, under conditions where male was either folded or unfolded by a denaturing agent. the results showed that both the covalent link and tertiary non-covalent interactions betwee ... | 1996 | 9005445 |
| 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 ... | 1997 | 9085128 |
| backbone dynamics of the major coat protein of bacteriophage m13 in detergent micelles by 15n nuclear magnetic resonance relaxation measurements using the model-free approach and reduced spectral density mapping. | the backbone dynamics of the major coat protein (gviiip) of the filamentous bacteriophage m13, solubilized in detergent micelles, have been studied using 15n nuclear magnetic resonance spectroscopy at three frequencies. motional parameters and overall and internal correlation times were derived with the model-free approach. it was also checked whether these parameters had to be modified due to anisotropic motion of the protein/micelle complex. reduced spectral density mapping was used to calcula ... | 1997 | 9092832 |
| distribution of sequence-dependent curvature in genomic dna sequences. | the distribution of inherent, sequence-dependent curvature was calculated for a number of prokaryotic (m. genitalium, h. influenzae, m. jannaschii), viral (adenovirus 2, equine herpes virus 1), phage (m13, lambda), eukaryotic (s. cerevisiae) and mitochondrial genomes as well as e. coli and human genomic fragments. the genomic averages are in the range of 6-8 degrees/helical turn and only about 20% of dna is curved less than 3 degrees/helical turn. the prokaryotes and phages appear to have a cons ... | 1997 | 9109388 |
| development of hiv-1 protease expression methods using the t7 phage promoter system. | new and simple human immunodeficiency virus type 1 (hiv-1) protease expression methods in escherichia coli were developed using the t7 phage promoter system. in order to suppress leaky hiv-1 protease expression under the control of the t7 polymerase, two new methods were tested. one involved the introduction of supplementary t7 promoter regions into host cells [e. coli bl-21 (de3)] containing the hiv-1 protease gene under the control of the t7 promoter. it was expected that the supplementary t7 ... | 1997 | 9114515 |
| comparison of four molecular typing methods for evaluating genetic diversity among candida albicans isolates from human immunodeficiency virus-positive patients with oral candidiasis. | candida albicans strain delineation by karyotyping. noti restriction pattern analysis, hybridization with specific probe 27a, and pcr fingerprinting with the phage m13 core sequence were performed with 30 isolates from the oral cavities of 30 human immunodeficiency virus (hiv)-infected patients and 8 reference strains. within the panel of clinical isolates, 20 were geographically related, although 10 isolates were susceptible to fluconazole and 10 isolates were resistant to fluconazole. the rema ... | 1997 | 9157142 |
| 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 ... | 1997 | 9177746 |
| a selection system to study protein-rna interactions: functional display of hiv-1 tat protein on filamentous bacteriophage m13. | the transactivator protein (tat) of the human immunodeficiency virus (hiv) is a key regulatory protein in the viral replication cycle and belongs to the rna binding proteins of the arginine-rich motif (arm) family. very little is known about their mechanism of rna recognition. to study the principles of rna-protein recognition we constructed a system to display hiv-1 tat on the surface of the filamentous bacteriophage m13. hiv-1 tat (1-72) and a mutant tat lacking five cysteine residues were clo ... | 1997 | 9207243 |
| mutations in the siv env and the m13 lacza gene generated in vitro by reverse transcriptases and dna polymerases. | to investigate the accuracy of retroviral in vitro dna replication we have examined with two fidelity assays the reverse transcriptases (rts) from sivagm, hiv-1, momlv as well for comparison the klenow fragment from e. coli and dna polymerase a from calf-thymus. these forward mutation assays measured the loss of bacteriophage m13 lacza gene function by mutations. in the envlacza assay frameshift mutations occurring during polymerisation of a 176 b long simian immunodeficiency virus (siv) envelop ... | 1997 | 9229004 |
| use of random amplification of polymorphic dna (rapd) and pcr-fingerprinting for genotyping a scedosporium prolificans (inflatum) outbreak in four leukemic patients. | four isolates of the pathogenic fungus scedosporium prolificans (inflatum), causing a previously reported nosocomial outbreak in four leukemic patients, were typed by random amplification of polymorphic dna (rapd) with two different 10-mer primers and pcr-fingerprinting with the core sequence of phage m13 as a single primer. both techniques allowed 10 additional clinical isolates of scedosporium prolificans from different areas of spain, including scedosporium prolificans ncpf 2884, to be classi ... | 1997 | 9236303 |
| a combinatorial method for constructing libraries of long peptides displayed by filamentous phage. | we describe the construction and screening of a random peptide library displayed by filamentous phage. the peptides are expressed in multiple copies on the filamentous phage m13 as amino-terminal fusions with the major coat protein, the product of gene viii. these libraries are efficiently screened for reactive peptides, using a combination of panning in solution followed by a plaque lift assay. advantages of this system are that both high- and low-affinity phage clones are simultaneously identi ... | 1995 | 9237193 |
| obtaining a family of high-affinity, high-specificity protein inhibitors of plasmin and plasma kallikrein. | human lipoprotein-associated coagulation inhibitor (laci) is a serum protein containing three kunitz domains. we displayed the first domain (laci-d1) on the iii protein of phage m13 and made libraries of this domain. we iteratively varied 13 residues in the region corresponding to the bpti-trypsin interface and selected for binding to human plasmin (pla) and human plasma kallikrein (pkal). for pla, our first-round best binder, epi-p211, had kd = 2 nm. using information from the first selection, ... | 1996 | 9238642 |
| 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 ... | 1997 | 9247162 |
| 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 ... | 1997 | 9260966 |
| 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 ... | 1997 | 9261557 |
| potential involvement of both type i and type ii mechanisms in m13 virus inactivation by methylene blue photosensitization. | we have investigated the mechanism of virus photoinactivation with methylene blue (mb) by conducting deuterium oxide (d2o), azide ion (n3-) and oxygen-dependent studies. inactivation of m13 bacteriophage and singlet oxygen (1o2) generation by mb photosensitization were irradiation dose dependent. inactivation of m13 was enhanced by d2o and inhibited by n3-, suggesting that 1o2 participates in m13 inactivation by mb photosensitization. however, n3- did not inhibit m13 inactivation completely. on ... | 1997 | 9277138 |
| 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 ... | 1997 | 9315865 |
| improving the copy numbers of antibody fragments expressed on the major coat protein of bacteriophage m13. | antibody fragments have been expressed on the major coat protein of filamentous phage using a gene viii expression system, but with low copy numbers (averaging 0.2 fab/phage). | 1996 | 9373312 |
| monoclonal antibody against piii of filamentous phage: an immunological tool to study piii fusion protein expression in phage display systems. | a monoclonal antibody directed against the gene 3 product (piii) of filamentous phage m13 was produced to study piii-fusion protein expression in e. coli and its incorporation in the phage capsid. the protein was gel-purified from e. coli expression cultures harboring the genetic information of piii under the control of an inducible lac promoter. to study piii-fusion protein expression, phage display systems were applied in which either the whole piii or the c-terminal half was used (mccafferty ... | 1995 | 9373333 |
| [characterization of plasmids which mediate resistance to multiple antibiotics in gram-negative bacteria of nosocomial origin]. | the genetic and molecular mechanisms involved in antimicrobial resistance of 10 strains of gramnegative bacilli (1 serratia marcescens; 2 escherichia coli; 1 proteus mirabilis; 4 klebsiella pneumoniae; 1 enterobacter cloacae y 1 alcaligenes faecalis), isolated from adult patients with nosocomial pulmonary infection at the in-patient facilities of the university hospital of los andes, mérida, venezuela, have been studied. | 1997 | 9376400 |
| 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 ... | 1997 | 9378120 |
| 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 ... | 1997 | 9399489 |
| 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 ... | 1997 | 9434182 |
| 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 ... | 1998 | 9443386 |
| 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 ... | 1997 | 9450488 |
| [the genetic typing of strains in the genus francisella using universal probes]. | the determination of the genetic relationship of bacteria of the genus francisella and their differentiation is one of the topical tasks of the epidemiology and infectology of f. tularensis, the causative agent of tularemia, belonging to this genus. to solve this task, investigation was carried out with a view to the determine the possibility of the genomic typing of francisella. genomic typing was based on the use of the hybridization of fragments of francisella chromosomal dna, split by restri ... | 1997 | 9460874 |
| 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 ... | 1998 | 9461610 |