Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| new versatile cloning and sequencing vectors based on bacteriophage m13. | a new pair of cloning and sequencing vectors based on bacteriophage m13mp7 has been developed. these vectors (m13tg130 and m13tg131) contain, in addition to the ecori, bamhi, hindiii, smai, sali and psti sites present in other vectors [cf., m13mp8 and m13mp9, messing and vieira, gene 19 (1982) 269-276], unique restriction recognition sequences for the enzymes ecorv, kpni, sphi, ssti and xbai. a restriction site for the enzyme bglii has been incorporated into the polylinker region of one of the v ... | 1983 | 6323254 |
| use of m13mp phages to study gene regulation, structure and function: cloning and recombinational analysis of genes of the salmonella typhimurium histidine operon. | a restriction map was determined for a phi 80 lambda dhis transducing phage dna carrying the salmonella typhimurium histidine operon. dna fragments containing the promoter/regulatory region and the first two structural genes of the histidine operon (hisogd) were identified by their ability to direct regulated synthesis of histidinol dehydrogenase (product of hisd) in a coupled in vitro protein synthesizing system. a 3.1-kb sali-ecori restriction fragment containing the hisogd region, was subclon ... | 1983 | 6323256 |
| in vitro generation of specific deletions in dna cloned in m13 vectors using synthetic oligodeoxyribonucleotides: mutants in the 5'-flanking region of the yeast alcohol dehydrogenase ii gene. | deletion mutants are particularly useful in defining the boundaries of noncoding genetic functions. such mutants can be precisely generated using synthetic oligodeoxyribonucleotides as mutagens. in this paper we describe the application of this method to recombinant dna cloned in a phage m13-derived vector. the mutagenic oligodeoxyribonucleotides, 20 and 21 nucleotides in length, were used to delete a tract of 20 da-dt base-pairs and an adjacent 22 base-pair perfect dyad from the adr3 locus, the ... | 1984 | 6324118 |
| screening recombinant phage m13 plaques with rna probes; a one-step procedure which identifies clones containing either of the complementary dna strands. | we describe a method for detecting specific dna sequences cloned in m13 phage vectors, based on the procedure of woo (in wu, r., methods in enzymology, vol. 68, academic press, new york, 1979, pp. 389-395). m13 plaques are adsorbed to a nitrocellulose filter that has been pre-saturated with bacteria. the filter is incubated on an agar plate to amplify the phage; the dna is alkali-denatured and then hybridized with a radioactive rna probe. unlike standard procedures, this method detects and disti ... | 1984 | 6325302 |
| some extrachromosomal circular dnas containing the alu family of dispersed repetitive sequences may be reverse transcripts. | a 300 base-pair (bp) size class of small polydisperse circular dna (spcdna) isolated from the bsc-1 line of african green monkey kidney cells was cleaved with the restriction endonuclease sau3a, and the resulting fragments (100 to 200 bp) were cloned in bacteriophage m13 mp7. the nucleotide sequence of each of 24 clones containing dna sequences homologous to the alu family of mobile, dispersed, repetitive elements was then determined. analysis of these sequences revealed that many, and perhaps a ... | 1984 | 6325707 |
| initiation and termination signals for transcription in bacteriophage m13. | transcription of the infrequently expressed phage m13 genome domain, comprising genes iii, vi, i and iv, has been studied in detail by hybridization and s1-nuclease mapping studies. the contiguous genes iii and vi are transcribed via an 1800 nucleotide-long rna molecule that is initiated at a promoter which overlaps with the rho-independent termination signal between genes iii and viii. its synthesis is terminated at a rho-dependent terminator in the proximal part of gene i. transcription of gen ... | 1984 | 6328409 |
| interference between m13 and orim13 plasmids is mediated by a replication enhancer sequence near the viral strand origin. | the origin of replication for the viral strand of bacteriophage m13 dna is contained within a 507 base-pair intergenic region of the phage chromosome. the viral strand origin is defined as the specific site at which the m13 gene ii protein nicks the duplex replicative form of m13 dna to initiate rolling-circle synthesis of progeny viral dna. using in vitro techniques we have constructed deletion mutations in m13 dna at the unique avai site which is located 45 nucleotides away on the 3' side of t ... | 1984 | 6332917 |
| m13 procoat and a pre-immunoglobulin share processing specificity but use different membrane receptor mechanisms. | bacteriophage m13 procoat is accurately processed to transmembrane coat protein by salt-washed or n-ethylmaleimide-treated rough microsomes from dog pancreas. these treatments inhibit the processing of eukaryotic secreted protein precursors. m13 procoat can assemble into dog pancreas microsomes post-translationally. thus, the microsomal proteins needed for assembly may be determined by the nature of the precursor protein itself. these results, and our finding that the mouse igg kappa chain fragm ... | 1983 | 6344069 |
| the use of bacteriophage m13 carrying defined fragments of the escherichia coli glta gene to determine the location and structure of the citrate synthase promoter region. | the glta gene from escherichia coli, which encodes citrate synthase, has been located on a 3.24 kb hindiii/ecorl restriction fragment. this region contains one restriction site for bamhl and two for bglii. defined restriction fragments from this region were cloned into suitably cleaved replicative form m13mp8 and m13mp9. the recombinants (m13gtla1 leads to 10) were isolated as single stranded dna and characterised on the basis of molecular weight and dna sequence. the single stranded dna was con ... | 1983 | 6355771 |
| in vitro deletional mutagenesis for bacterial production of the 20,000-dalton form of human pituitary growth hormone. | the 20,000-dalton (20k) variant form of human growth hormone (hgh) present in extracts from pituitary glands differs from the major form of hgh (22k, 191 amino acids) by the deletion of amino acid residues 32-46. using oligonucleotide-mediated mutagenesis, the dna coding for these amino acids was deleted from the gene previously constructed by us (goeddel et al., 1979) for microbial hgh production. the dna to be deleted was looped out by the annealing of a synthetic oligodeoxyribonucleotide to t ... | 1983 | 6357679 |
| transitory recombination between plasmid phv33 and phage m13. | plasmid phv33 and phage m13 which have no homology exceeding 13 bp, combine in escherichia coli cells. the chimeric genome is encapsidated in phage proteins and injected into a recipient cell, where it decombines to regenerate the two parental genomes. we call this combination-decombination process 'transitory recombination'. | 1983 | 6365530 |
| different base/base mismatches are corrected with different efficiencies by the methyl-directed dna mismatch-repair system of e. coli. | the efficiency of methyl-directed dna mismatch-repair of e. coli acting in vivo on heteroduplex genomes of phage m13 was found to be strongly dependent on the nature of the base/base mismatch to be corrected. three efficiency classes were characterized:high (t/g, c/a and g/g); intermediate (a/a); and low (g/a, a/g, t/t, c/c, c/t and t/c). methyl-directed dna mismatch repair was lost completely for any type of mismatch in strains carrying either mutl or muts mutations. data obtained with a muth m ... | 1984 | 6386179 |
| on the processivity of dna replication. | in this paper we describe the nature and importance of processive enzymatic reactions in biological processes. a model is set up to describe the processive synthetic process in dna replication, and experiments are presented to define and test the model, using the components of the t4 phage-coded five-protein (in vitro) dna replication system of alberts. nossal and coworkers. these experiments are performed either with a homogeneous oligo dt-poly da primer-template system, or with a natural prime ... | 1983 | 6400896 |
| rapid mutational analysis of regulatory loci in escherichia coli k-12 using bacteriophage m13. | a derivative of bacteriophage m13mp8 , designated m13mp8 /p, was prepared in which the promoter and nh2-terminal codons of bacterial genes may be fused to a portion of beta-galactosidase, resulting in an easily scorable phenotype. because transcription from the inserted promoter remains responsive to the host regulatory system, it is simple to screen mutagenized phage for isolates with aberrant regulatory phenotypes and to determine the mutational changes by dideoxy sequence analysis. the feasib ... | 1984 | 6427775 |
| replication origin of the bacillus subtilis chromosome determined by hybridization of the first-replicating dna with cloned fragments from the replication origin region of the chromosome. | the replication origin (ori) on the bacillus subtilis genome was determined by the hybridization between the first-replicating dna region and the cloned fragments from the ori region. the first-replicating dna region was labeled specifically by [3h]thymidine in the presence of an inhibitor for dna polymerase during a synchronous initiation of the chromosomal replication by germinating spores starved for thymine, and isolated by a sucrose density gradient centrifugation. most of the labeled dna m ... | 1984 | 6439606 |
| effect of spermine on interaction of dna polymerase alpha from the loach (misgurnus fossilis) eggs with dna. | polyamines (putrescine, spermidine and spermine) cause a marked increase in the activity of the loach misgurnus fossilis dna polymerase alpha on activated (gapped) dna. the stimulatory effect increases in the order: putrescine, spermidine, spermine. kinetic analysis shows that spermine does not change the affinity of the polymerase for dttp, but it decreases the enzyme affinity for dna. the apparent km of the polymerase for activated dna progressively increases from 14 to 1200 microm (nucleotide ... | 1984 | 6548155 |
| structure of murine complement component c3. i. nucleotide sequence of cloned complementary and genomic dna coding for the beta chain. | the nucleotide sequence coding for the beta chain of murine c3 was determined from cloned cdna and genomic dna fragments. sonicated subfragments were randomly inserted into the bacteriophage m13 and sequenced using the dideoxynucleotide technique. each nucleotide was sequenced on average six times in these studies. the derived amino acid sequence includes a signal peptide and a tetra-arginine sequence between the beta and alpha subunits in the precursor polypeptide prepro-c3. together with the a ... | 1984 | 6548745 |
| cloned single- and double-stranded dna copies of potato spindle tuber viroid (pstv) rna and co-inoculated subgenomic dna fragments are infectious. | a set of monomeric and oligomeric potato spindle tuber viroid (pstv) specific dna forms representing complete dna copies of the circular pstv rna genome were constructed and cloned in plasmid pbr322 and bacteriophage m13. both single- and double-stranded pstv dnas are capable of initiating viroid replication in mechanically inoculated tomato plants where it normally proceeds via the rna-rna pathway without dna being involved. all dimeric and higher multimeric forms were infectious irrespective o ... | 1984 | 6549294 |
| a detailed mutational analysis of the eucaryotic trnamet1 gene promoter. | we have isolated phage m13 clones containing the x. laevis trnamet1 gene, each having one or a few c leads to t transitions in the trna coding sequence. nearly every g-c and c-g base pair in the tdna has been mutagenized. the importance of these altered nucleotides in transcription by rna polymerase iii has been assessed by injecting the cloned dnas into frog oocyte nuclei together with alpha-32p-gtp and measuring the synthesis of labeled trnamet1. several g-c and c-g base pairs in the structura ... | 1983 | 6552939 |
| splicing of adenovirus rna in a cell-free transcription system. | a soluble whole-cell extract prepared accurately from hela cells splices 2-3% of the rna transcribed from a dna template containing the first and second leader exons of late adenovirus rna. the spliced rna was detected by a sensitive technique using hybridization to a single-stranded phage m13 cdna clone, followed by binding to nitrocellulose filters. the identity of the spliced rna was established by rnase t1 and pancreatic rnase two-dimensional peptide mapping. the bond formed during the in vi ... | 1983 | 6577417 |
| fluorescence studies of the complex formation between the gene 5 protein of bacteriophage m13 and polynucleotides. | 1983 | 6601193 | |
| a 500-mhz proton nuclear magnetic resonance study of the structure and structural alterations of gene-5 protein-oligo(deoxyadenylic acid) complexes. | the complex of the gene-5 protein of bacteriophage m13 with octadeoxyadenylic acid [d(a)8] has been shown earlier to differ in various respects from the complex with polynucleotides [alma, n. c. m., harmsen, b. j. m., van boom, j. h., van der marel, g., & hilbers, c. w. (1982) eur. j. biochem. 122, 319-326]. in this paper the gene-5 protein-d(a)8 complex is compared with the complex formation between the gene-5 protein and a mixture of longer oligonucleotides, i.e., d(a)25-30. nuclear overhauser ... | 1983 | 6602628 |
| an efficient synthetic primer for the m13 cloning dideoxy sequencing system. | the deoxytetradecamer d(aaaacgacggccag) has been shown to be an excellent universal primer for sequence determination of dna cloned into the bacteriophage m13 mp7, mp8, and mp9 series. this new primer offers several advantages over others currently available and it has been used to define the cloning of hinf i fragments of bacteriophage s13 dna into the eco ri site of m13 mp7, utilizing the homologous complementary base pairing of the two restriction sites. of the four possible sequence derivati ... | 1982 | 6753962 |
| the yeast his3 promoter contains at least two distinct elements. | phenotypic analysis of 65 mutations indicates that the yeast his3 promoter is composed of at least two separate regions of dna. each is necessary, but neither is sufficient for wild-type levels of his3 expression. deletion mutations that destroy either promoter element express his3 poorly or not at all. the upstream element is located between 112 and 155 base pairs before the site of transcriptional initiation (nucleotides -112 to -155). a comparison of derivatives strongly suggests that the dow ... | 1982 | 6760196 |
| coat protein conformation in m13 filaments, i-forms and spheroids. | circular dichroism studies of the filamentous coliphage m13 were carried out to determine conformational changes in the major capsid protein (the b protein) that occur during contraction of the filaments to i-forms and spheroids. the alpha-helicity of the b protein is somewhat lower in the i-forms than in filaments and much lower in spheroids. this conformational change may explain the increased detergent and lipid solubility of both i forms and spheroids relative to filaments. | 1983 | 6847652 |
| proteins encoded near the adenovirus late messenger rna leader segments. | small fragments of adenovirus 2 dna cloned into the single-strand phage m13 were used to select adenoviral messenger rnas transcribed from the r-strand between map positions 16 and 30. cell-free translation of these mrnas produced proteins of 13.5k, 13.6k, and 11.5k, respectively encoded between the first and second segments of the tripartite major late leader, within the "i"-leader segment, and immediately preceding the third leader segment. partial sequence analysis of the 13.6k protein is con ... | 1983 | 6857999 |
| characterization and properties of a modified human interferon-alpha containing an additional 18 amino acids at the n-terminus. | a modified human interferon-alpha 2 was produced in escherichia coli cells infected with phage m13 mp7 containing an interferon-alpha gene. after purification by immunochromatography with the monoclonal antibody nk2, the n-terminal amino acid sequence was determined. the n-terminal methionine was absent but an additional sequence of 18 amino acids at the n-terminus was retained. the modified interferon-alpha 2 was indistinguishable from authentic interferon-alpha 2 in its ability to activate nat ... | 1983 | 6875519 |
| nucleotide-sequence heterogeneity and sequence rearrangements in influenza virus cdna. | double-stranded cdna has been synthesized from influenza virus rna and cloned into derivatives of the bacteriophage m13 for sequence analysis. the characterization of over 200 clones has permitted an analysis both of nucleotide sequence heterogeneity and of clones containing unusual rearrangements of sequence. heterogeneity, due to genetic variability in the rna population and to in vitro synthetic errors, was detected at the low level of one nucleotide difference per 3 700 nucleotides. by contr ... | 1981 | 6895362 |
| mechanisms of membrane assembly: effects of energy poisons on the conversion of soluble m13 coliphage procoat to membrane-bound coat protein. | the coat protein (gene 8 product) of coliphage m13 spans the host cell plasma membrane prior to its assembly into extruding virions. it is made as a soluble precursor, termed procoat, with an extra 23 nh2-terminal amino acid residues. we have examined the effect of metabolic poisons on the assembly of procoat into the plasma membrane and its proteolytic conversion to coat protein. protein synthesis and proline uptake were measured to assess the effect of each poison on cellular high-energy phosp ... | 1980 | 6928682 |
| replication of the plasmid pbr322 under the control of a cloned replication origin from the single-stranded dna phage m13. | the replication origins of viral and complementary strands of bacteriophage m13 dna are contained within a 507-nucleotide intergenic region of the viral genome. chimeric plasmids have been constructed by inserting restriction endonuclease fragments of the m13 intergenic region into the plasmid pbr322. replication of these hybrid plasmids, under conditions not permissive for the plasmid replicon, depends on specific segments of the m13 origin region and on the presence of m13 helper virus. thus m ... | 1980 | 6933512 |
| genes vi, vii, and ix of phage m13 code for minor capsid proteins of the virion. | the minor capsid proteins c and d from phage m13 have been characterized by differential amino acid labeling and amino-terminal sequence analysis. we demonstrate that d protein (mr 12,260) is the product of gene vi, whereas the c component is composed of the products of both gene vii (mr 3580) and gene ix (mr 3650). our data further show that the proteins of genes vi, vii, and ix are not subject to proteolytic processing but are packaged into mature virions as their primary translational product ... | 1981 | 6945579 |
| "nonrandom" dna sequence analysis in bacteriophage m13 by the dideoxy chain-termination method. | we describe a rapid "nonrandom" dna sequence analysis procedure that facilitates the nucleotide sequence determination of large contiguous regions of dna. the method consists of cloning a restriction endonuclease fragment of interest into bacteriophage m13 followed by construction of a series of nuclease bal-31 deletion mutants originating from a single site in m13 that is close to the dna insert. determination of the size of the deletion mutant is accomplished by hybridization to a complementar ... | 1982 | 6956859 |
| rna processing errors in patients with beta-thalassemia. | we have developed a method that permits rapid identification of the consequences of mutations that alter beta-globin rna processing in erythroid cells. s1 nuclease mapping techniques were used to analyze total bone marrow rna obtained fron 15 patients who are clinically homozygous for beta-thalassemia and from 5 patients with erythroid hyperplasia from other causes. this analysis was facilitated by the use of single-stranded uniformly labeled dna probes of high specific activity that were prepar ... | 1982 | 6956887 |
| nmr studies of the interaction of gene-v protein of bacteriophage m13 with oligonucleotides. | this paper describes the preparation of deuterated phenylalanine ([2h7]-phenylalanine) and the isolation of phage m13 encoded gene-v protein in which this deuterated amino acid was incorporated. using this protein spectral assignments of resonances in the aromatic region of the 1h-nmr spectrum of the gene-v protein have been made. furthermore the interaction of the gene-v protein with the tetranucleotide d(pc-g-c-g) and the hexanucleotide d(pc-g-c-g-c-g) was investigated. from the changes in the ... | 1980 | 6966158 |
| double-resonance experiments at 500 mhz on gene-5 protein and its complex with octadeoxyriboadenylic acid. | in this paper, a detailed description is presented of the aromatic part of the 500-mhz 1h nuclear magnetic resonance (nmr) spectrum of the helix-destabilizing gene-5 protein (gvp) encoded by the coliphage m13. as a result of the resolution obtained at 500 mhz, it was possible to perform selective decoupling and time-resolved selective overhauser experiments. the magnitudes of the observed overhauser effects compare favorably with magnitudes expected on the basis of theoretical calculations. thes ... | 1981 | 6974567 |
| 1h nmr studies of the binding of bacteriophage-m13-encoded gene-5 protein to oligo(deoxyadenylic acid)s of varying length. | the binding of gene-5 protein to oligo(deoxyadenylic acid)s varying in length from 2 to 16 nucleotides has been studied by titrating the protein with the oligonucleotides and recording the 1h nmr spectra at 360 mhz. to obtain information about the mode of binding of the protein the aromatic parts of the spectra have been analysed by performing spectral simulations, starting from the assignments obtained from nuclear overhausfer enhancements at 500 mhz [alma, n. c. m., harmsen, b. j. m., hull, w. ... | 1982 | 6977447 |
| synthesis, assembly into the cytoplasmic membrane, and proteolytic processing of the precursor of coliphage m13 coat protein. | 1980 | 6986388 | |
| purification and characterization of leader (signal) peptidase from escherichia coli. | many membrane proteins and secreted proteins are synthesized in precursor form with 15 to 30 additional nh2-terminal residues. these "leader peptides" (pre-pieces, signal peptides) are removed as these proteins cross or insert into cellular membranes. "leader peptidase" activities which catalyze this cleavage have been detected in crude extracts and found to be dependent on membrane fractions. we now describe a 6,000-fold purification of a leader peptidase from the membranes of uninfected escher ... | 1980 | 6995457 |
| procoat, the precursor of m13 coat protein, requires an electrochemical potential for membrane insertion. | the coat protein of coliphage m13 spans the host cell cytoplasmic membrane prior to its assembly into extruding virus. it is made as a soluble cytoplasmic precursor, termed "procoat," with 23 extra amino acid residues at the nh2 terminus. procoat binds to the cell membrane and is converted proteolytically to coat protein. when the electrochemical gradient of an infected cell is rapidly dissipated by uncouplers, procoat still binds to the plasma membrane but is not converted to coat. we report he ... | 1980 | 7001463 |
| expression of a dna strand initiation sequence of cole1 plasmid in a single-stranded dna phage. | in order to investigate initiation of h-strand (lagging strand) replication of the plasmid cole1, the origin region fragment (hae ii-e) of cole1 was inserted into the intergenic region of filamentous dna phage m13 and cloned. a site capable of promoting dna strand initiation on a single-stranded dna template has been detected on the l-strand (leading strand) of the cloned fragment. the site, named rri-1 rifampicin-resistant initiation), directs conversion of chimeric phage single-stranded dna to ... | 1980 | 7005899 |
| expression of bacteriophage m13 dna in vivo. isolation, identification and characterization of phage-specific mrna species. | 1980 | 7007041 | |
| leader peptidase is found in both the inner and outer membranes of escherichia coli. | many membrane proteins are synthesized as transient precursors with an nh2-terminal leader (or signal) peptide. during insertion of these proteins into the membrane, leader peptides are removed by leader peptidase. one such enzyme has been detected in detergent extracts of escherichia coli membranes and extensively purified using as an assay the removal of the leader sequence of procoat, the precursor of the major coat protein of bacteriophage m13. we now report that this leader peptidase is fou ... | 1981 | 7009614 |
| structure of the neuraminidase gene in human influenza virus a/pr/8/34. | the complete structure of the neuraminidase gene in influenza a/pr/8/34 has been determined by cloning into the bacteriophage m13 and sequencing with dideoxynucleotide chain terminators. the gene is 1,413 nucleotides long, codes for a protein of 454 amino acids and has five potential glycosylation sites. we suggest that the neuraminidase, unlike the influenza haemagglutinin, is oriented with its n-terminus buried in the viral membrane. | 1981 | 7010182 |
| procoat, the precursor of m13 coat protein, inserts post-translationally into the membrane of cells infected by wild-type virus. | in growing cells infected by wild-type coliphage m13, the synthesis of procoat protein is completed before it inserts into the plasma membrane ane is converted to coat protein. | 1981 | 7014926 |
| membrane assembly from purified components. i. isolated m13 procoat does not require ribosomes or soluble proteins for processing by membranes. | the coat protein of coliphage m13 is an integral protein of the host-cell cytoplasmic membrane prior to its assembly into virions. it is initially synthesized as procoat, a soluble precursor with a 23 amino acid leader sequence at its amino terminus. 35s-labeled procoat accumulates during an in vitro translation reaction that contains 35s-methionine and rna from m13-infected cells. radiochemically pure procoat has been isolated from in vitro translation reactions by extraction into an organic so ... | 1981 | 7026042 |
| membrane assembly from purified components. ii. assembly of m13 procoat into liposomes reconstituted with purified leader peptidase. | the major coat protein of coliphage m13 is an integral protein of the e. coli plasma membrane prior to its assembly into new virus particles. it is generated from its precursor, procoat, by a membrane-bound leader peptidase. we now describe the reconstitution of a highly purified preparation of this enzyme into vesicles of e. coli phospholipids. these vesicles bind procoat made in vitro and procoat isolated from in vitro synthesis. both the crude and the purified substrates were converted post-t ... | 1981 | 7026043 |
| radiation-induced base substitution mutagenesis in single-stranded dna phage m13. | 1981 | 7029307 | |
| effects of dna base analogs on transcription termination at the tryptophan operon attenuator of escherichia coli. | we have devised a method to specifically incorporate deoxyribonucleotide base analogs in vitro into either strand of the tryptophan (trp) operon attenuator region, using primed synthesis on bacteriophage m13 derivatives carrying cloned trp attenuator dna. we have employed these techniques to extend previous studies implicating both rna-rna and rna-dna interactions in transcription termination in an attempt to determine the nature of the contribution from the template dna molecule in termination ... | 1982 | 7041118 |
| the biosynthesis of membrane-bound m13 coat protein. energetics and assembly intermediates. | the major coat protein of bacteriophage m13 spans the plasma membrane of infected cells prior to its assembly into extruding virus. it is initially made as a precursor, termed procoat, with a 23-residue leader sequence at its nh2 terminus. procoat is found bound to the inner surface of the plasma membrane. the electrical potential of the cell membrane is required for procoat insertion and conversion to coat protein, although the order of these events has been unknown. we now report studies of th ... | 1982 | 7042715 |
| evidence for two genetically distinct dna primase activities specified by plasmids of the b and i incompatibility groups. | plasmid colib-p9 of the i alpha incompatibility group is known to encode a dna primase that acts in the conjugal transfer of the plasmid and can substitute for mutant dnag gene product in vegetative replication of the escherichia coli chromosome. the relevant genetic determinant (sog) has previously been cloned into a small multicopy vector plasmid. prototype incb plasmid r16 also suppresses host dnag mutations. the equivalent gene(s) (pri) of r16 was cloned into plasmid pbr325 and shown by filt ... | 1982 | 7045070 |
| mutability of bacteriophage m13 by ultraviolet light: role of pyrimidine dimers. | the role of pyrimidine dimers in mutagenesis by ultraviolet light was examined by measuring the uv-induced reversion of six different bacteriophage m13 amber mutants for which the neighboring dna sequences are known. the mutational response at amber (tag) codons preceded by a guanine or adenine (where no pyrimidine dimer can be formed) were compared with those preceded by thymine or cytosine (where dimer formation is possible). equivalent levels of uv-induced mutagenesis were observed at both ki ... | 1982 | 7048024 |
| [transfection of bacterial cells by dna of phage m13 entrapped in phospholipid vesicles]. | the possibility of transfection of bacterial cells by phage m13 dna entrapped into phospholipid vesicles (liposomes) has been studied. two types of liposomes differing in size were used. entrapped dna was transferred by liposomes into ca2+-treated e. coli cells. efficiency of the transfection in the case of small (ca. 400 a) liposomes was 2--3 orders of magnitude higher than that of free dna extracted from such liposomes. | 1982 | 7048068 |
| 19f nuclear magnetic resonance studies of the coat protein of bacteriophage m13 in synthetic phospholipid vesicles and deoxycholate micelles. | the nonlytic, filamentous coliphage m13 offers an excellent model system for the study of membrane-protein interactions. we prepare derivatives of the protein containing fluorine-labeled amino acids and use 19f nuclear magnetic resonance (nmr) to study the protein in both deoxycholate micelles and phospholipid vesicles. we have previously described the in vivo preparation of an m-fluorotyrosyl derivative of m13 coat protein and also a method for incorporation of high levels of this protein into ... | 1982 | 7055622 |
| bilayer acyl chain dynamics and lipid-protein interaction: the effect of the m13 bacteriophage coat protein on the decay of the fluorescence anisotropy of parinaric acid. | nanosecond fluorescence polarization anisotropy decay is used to determine the effect of the bacteriophage m13 coat protein on lipid bilayer acyl chain dynamics and order. the fluorescent acyl chain analogues cis- and trans-parinaric acid were used to determine the rate and extent of the angular motion of acyl chains in liquid crystalline (39 degrees c) dimyristoylphosphatidylcholine bilayers free of coat protein or containing the coat protein at a protein:lipid ratio of 1:30. subnanosecond time ... | 1982 | 7055623 |
| beta-thalassemia in a kurdish jew. single base changes in the t-a-t-a box. | we recently described a "non-random" sequencing procedure for dna inserts in bacteriophage m13 using bal 3 nuclease and the dideoxy chain termination method (poncz, m., solowiejczyk, d., ballantine, m., schwartz, e., and surrey, s. (1982) proc. natl. acad. sci. u. s. a., in press). using this procedure, we have determined the nucleotide sequence of a cloned human beta-globin gene from a kurdish jew with beta +-thalassemia major. comparison with the previously reported human beta-globin gene sequ ... | 1982 | 7076659 |
| circular single stranded phage m13-dna as a template for dna synthesis in protein extracts from xenopus laevis eggs: evidence for a eukaryotic dna priming activity. | unfractionated protein extracts from activated xenopus laevis eggs contain all functions required for the chain elongation reactions in replicative dna synthesis (a.richter, b. otto and r.knippers, 1981, nucl.ac.res. 9, 3793-3807). in order to further explore the dna synthesizing capacity of this in vitro system and to obtain information on the dna priming activity in these extracts single stranded phage m13-dna was used as template for in vitro dna synthesis. the main results of this investigat ... | 1982 | 7145711 |
| filamentous bacteriophage contract into hollow spherical particles upon exposure to a chloroform-water interface. | the bacteriophage m13 is a 1 micrometer long filament consisting of a circular single-stranded dna loop firmly held within a tubular protein and capsid. we report here that exposure to a chloroform-water interface initiates a 20 fold contraction of each filament into a hollow protein sphere. in these 0.04 micrometer diameter particles, termed m13 "spheroids," two thirds of the dna is apparently extruded through a hole in the wall of the spheroid; the portion of dna remaining inside the shell cen ... | 1981 | 7226228 |
| membrane assembly: posttranslational insertion of m13 procoat protein into e. coli membranes and its proteolytic conversion to coat protein in vitro. | the major coat protein (gene 8 product) of bacteriophage m13 is an integral membrane protein during infection of host cells. it is synthesized as a larger precursor (procoat) with a leader sequence of 23 amino acids at its amino terminus. in vivo studies have shown that procoat only inserts into the host-cell plasma membrane after its synthesis is completed. we now demonstrate that procoat can post-translationally insert into inverted cytoplasmic membrane vesicles from e. coli and can be process ... | 1981 | 7237555 |
| deletion analysis of the cloned replication origin region from bacteriophage m13. | a cloned 270-nucleotide fragment from the origin region of the m13 duplex replicative form dna confers an m13-dependent replication mechanism upon the plasmid vector pbr322. this m13 insert permits m13 helper-dependent replication of the hybrid plasmid in pola cells which are unable to replicate the pbr322 replicon alone. using in vitro techniques, we have constructed several plasmids containing deletions in the m13 dna insert. the endpoints of these deletions have been determined by dna sequenc ... | 1981 | 7288922 |
| mechanism of coliphage m13 contraction: intermediate structures trapped at low temperatures. | the filamentous coliphage m13 can be transformed into a spherical particle (termed spheroid) by exposure to an interface of water and slightly polar but hydrophobic solvent such as chloroform-water at 24 degrees c. we report here that exposure of m13 filaments to a chloroform-water interface at 2 degrees c trapped the phage particles in forms morphologically intermediate to filaments and spheroids. these structures were rods 250 nm long and 15 nm wide, and each had a closed, slightly pointed end ... | 1981 | 7321105 |
| genes vi, vii and ix of bacteriophage m13: identification of their products as minor capsid proteins. | in earlier work, two new minor capsid proteins of molecular weight 3500 (c-protein) and 11,500 (d-protein) were detected in m13 virions. to determine their genetic origin, differential amino acid labeling, amino acid analysis and edman degradation analysis were performed on these proteins. the data demonstrate that d-protein is the product of gene vi whereas c-protein is composed of both the proteins specified by gene vii and the recently discovered gene ix. by selective labeling with arginine, ... | 1981 | 7330055 |
| a clear-plaque mutation of bacteriophage m13 affects the regulation of viral dna synthesis. | a clear-plaque mutation (c2) of bacteriophage m13 has been shown to affect the regulation of viral dna synthesis. this mutation increases the amount of the duplex replicative form dna per cell while decreasing the synthesis of viral single strands. the relative synthesis of the m13 gene 5 protein is approximately half that observed in wild-type infections, suggesting that the effect of the c2 mutation on the regulation of viral dna synthesis is a result of reduced expression of gene 5. | 1980 | 7365875 |
| nucleotide sequences in bacteriophage f1 dna: nucleotide sequence of genes v, vii, and viii. | the sequence of nucleotides comprising genes v, vii, and viii of bacteriophage f1 was determined. the sequence was found to differ from that of the corresponding region of the related fd genome by eight base substitutions in gene v and one in gene viii. the structure of gene vii was completely conserved between these two viruses and was identical to that of bacteriophage m13. both transitions and transversions were found in cases where bases were substituted, but all substitutions were in the th ... | 1980 | 7373712 |
| phage display selection of ligand residues important for src homology 3 domain binding specificity. | the src homology 3 (sh3) domain is a 50-aa modular unit present in many cellular proteins involved in intracellular signal transduction. it functions to direct protein-protein interactions through the recognition of proline-rich motifs on associated proteins. sh3 domains are important regulatory elements that have been demonstrated to specify distinct regulatory pathways important for cell growth, migration, differentiation, and responses to the external milieu. by the use of synthetic peptides, ... | 1995 | 7479908 |
| virucidal short wavelength ultraviolet light treatment of plasma and factor viii concentrate: protection of proteins by antioxidants. | the use of solvent/detergent mixtures and various forms of heat treatment to inactivate viruses has become widespread in the preparation of blood derivatives. because viruses that lack lipid envelopes and/or are heat resistant, eg, hepatitis a virus (hav) or parvovirus b19 may be present, the use of two methods of virus elimination that operate by different mechanisms has been advocated. we now report on short wavelength ultraviolet light (uvc) irradiation for virus inactivation and enhancement ... | 1995 | 7492794 |
| molecular genetic analysis of a thioredoxin gene from thiobacillus ferrooxidans. | the thiobacillus ferrooxidans thioredoxin gene, trxa, was isolated by its ability to complement an escherichia coli gsha trxa mutant which was otherwise unable to grow on minimal medium lacking glutathione. the t. ferrooxidans thioredoxin also enabled the in vivo reduction by e. coli of methionine sulfoxide to methionine, as well as the in vitro reduction of insulin. when present in e. coli, the t. ferrooxidans thioredoxin supported the replication of phage t7, but not the growth of phage m13. t ... | 1995 | 7496529 |
| identification of antigenic sites on the na+/k(+)-atpase beta-subunit: their sequences and the effects of thiol reduction upon their structure. | in contrast to the catalytic (alpha) subunit of the na+/k(+)-atpase holoenzyme, the glycoprotein (beta) subunit has proven to be a poor antigen for monoclonal antibody (mab) production. however, in this work six mabs directed against the beta-subunit of the lamb kidney holoenzyme have been isolated. these mabs all recognize the holoenzyme, but their 'in solution' binding affinities for deglycosylated enzyme or isolated beta are generally at least 10-fold higher. species specificity mapping, anti ... | 1994 | 7521214 |
| tendamistat as a scaffold for conformationally constrained phage peptide libraries. | the alpha-amylase inhibitor tendamistat (hoe-467), a 74 amino acid beta-sheet protein from streptomyces tendae has been expressed on the surface of the filamentous bacteriophage m13. phage displaying tendamistat inhibit the hydrolysis of starch by alpha-amylase, indicating that the displayed protein is functional. the displayed tendamistat has been used as a molecular scaffold for the presentation of constrained random peptides. two loops, comprising residues 38 to 40 and 60 to 65 of tendamistat ... | 1995 | 7542349 |
| nmr studies of the major coat protein of bacteriophage m13. structural information of gviiip in dodecylphosphocholine micelles. | the membrane-bound form of the major coat protein (gviiip) of bacteriophage m13 has been studied using nuclear magnetic resonance spectroscopy. as membrane mimetics, we used dodecylphosphocholine (dodpcho) detergent micelles to solubilize the protein. we were able to nearly completely assign all resonances of the protein solubilized in dodpcho micelles by using both homonuclear and heteronuclear multidimensional experiments. based on the patterns of the nuclear overhauser enhancements and the ch ... | 1995 | 7556198 |
| refined solution structure of the tyr41-->his mutant of the m13 gene v protein. a comparison with the crystal structure. | the three-dimensional solution structure of mutant tyr41-->his of the single-stranded dna binding protein encoded by gene v of the filamentous bacteriophage m13 has been refined in two stages. the first stage involved the collection of additional noe-based distance constraints, which were then used in eight cycles of back-calculations and structure calculations. the structures of the gene v protein dimers were calculated using simulated annealing, employing restrained molecular dynamics with a g ... | 1995 | 7556200 |
| single-stranded dna binding protein encoded by the filamentous bacteriophage m13: structural and functional characteristics. | the single-stranded dna binding protein, or gene v protein (gvp), encoded by gene v of the filamentous bacteriophage m13 is a multifunctional protein that not only regulates viral dna replication but also gene expression at the level of mrna translation. it furthermore is implicated as a scaffolding and/or chaperone protein during the phage assembly process at the hostcell membrane. the protein is 87 amino acids long and its biological functional entity is a homodimer. in this manuscript a short ... | 1994 | 7565651 |
| characterization of the extracellular lipase, lipa, of acinetobacter calcoaceticus bd413 and sequence analysis of the cloned structural gene. | the extracellular lipase from acinetobacter calcoaceticus bd413 was purified to homogeneity, via hydrophobic-interaction fast performance liquid chromatography (fplc), from cultures grown in mineral medium with hexadecane as the sole carbon source. the enzyme has an apparent molecular mass of 32 kda on sds-polyacrylamide gels and hydrolyses long acyl chain p-nitrophenol (pnp) esters, like pnp palmitate (pnpp), with optimal activity between ph 7.8 and 8.8. additionally, the enzyme shows activity ... | 1995 | 7596283 |
| efficient incorporation of anti-hiv deoxynucleotides by recombinant yeast mitochondrial dna polymerase. | saccharomyces cerevisiae mtdna polymerase, isolated as a single 135-kda recombinant polypeptide, showed high processivity and a capacity of use poly(da).oligo(dt), poly(ra).oligo(dt), or primed bacteriophage m13 dna as a template. in a primer extension assay, the enzyme exhibited an intrinsic 3'-5'-exonuclease activity. by optimizing the polymerization reaction conditions, apparent km and vmax values could be determined for the incorporation of dttp, 2'-3'-dideoxy-ttp (ddttp), 3'-azido-ttp (aztt ... | 1995 | 7642550 |
| [the dna damage by photodynamic effects of hematoporphyrin derivatives (hpd)]. | photodynamic effects of hpd has been used for cancer therapy (pdt) successfuly. the biological mechanism of the pdt was studied by using dna as a target in this work. after treatment of phage m13 dna with hpd + light the molecule of the dna kept intact in te buffer. but the smear bands by puting dna in 0.1 mol/l naoh at 90 degrees c and separating with electrophoresis denoted the degradation of dna at the alkali-labile sites of phosphodiester bond in which the groups of the bases were photooxydi ... | 1995 | 7656396 |
| packing of coat protein amphipathic and transmembrane helices in filamentous bacteriophage m13: role of small residues in protein oligomerization. | filamentous bacteriophage m13, an important cloning and phage display vector, is encapsulated by ca 2700 copies of its 50-residue major coat protein (gene 8). this protein occurs as a membrane protein while stably inserted into its e. coli host inner membrane, and as a coat protein upon assembly and packing onto phage dna in the lipid-free virion. to examine the specific protein-protein interactions underlying these processes, we used a combination of randomized and saturation mutagenesis of the ... | 1995 | 7666434 |
| chaperonin assisted phage display of antibody fragments on filamentous bacteriophages. | we have used the groe chaperonins to assist in the packing of a new phage display vector, pexmide3. titers of the packed phagemid increased almost 200-fold from approximately 4 x 10(11) cfu/ml, without coexpression of the groe proteins, to approximately 7 x 10(13) cfu/ml with their coexpression. equal titers of non-assisted and assisted phagestocks exhibited the same antigen specificity and elisa reactivity, indicating the same frequency of displayed fab-fragments. while the diversity of antibod ... | 1993 | 7682084 |
| design of specific immunogens using filamentous phage as the carrier. | earlier, we developed an expression vector allowing exposure of short peptides on the surface of bacteriophage m13. it was used to obtain a recombinant phage carrying an antigenic determinant of hiv1 p17 gag protein. immunoglobulin elicited by immunizing rabbits with the phage reacted with the 17-kda core protein of the virus and with its polyprotein precursor, p55, on western blots of hiv1 viral proteins. the results of present experiments may be useful in vaccine development. | 1993 | 7685305 |
| [preparation of a specific immunogen based on bacteriophage m13]. | earlier we developed an expression vector on the basis of bacteriophage m13 allowing the exposure of short peptides on the virion surface. it was used to obtain a recombinant phage carrying the antigenic determinant of hivi gag protein p17. this phage was tested as immunogen in rabbits. it was shown by elisa that ig against the fusion phage reacted with the 17-kda core protein of the virus and with its polyprotein precursor p55 on strips activated by the transfer of hivi viral proteins. these da ... | 1993 | 7686250 |
| mutagenic and recombinagenic effects of diethylstilbestrol quinone. | estrogens are believed to be major contributors to many cancers of the human female genital tract, but the mechanism of their carcinogenic action is not well-understood. while a tumor-promoting role for estrogens is well-supported, whether they also act as tumor initiators has remained controversial. here, we have sought to examine the mutagenic potential of diethylstilbestrol, a synthetic estrogen that is a powerful carcinogen in hamsters, and is suspected to be a human carcinogen. phage m13 si ... | 1993 | 7690889 |
| evaluation of antibodies fused to minor coat protein iii and major coat protein viii of bacteriophage m13. | a gene coding for an anti-(2-phenyl-5-oxazolone) single-chain fv antibody (ab) fragment (anti-phox scfv) was cloned in-frame into phagemid vectors upstream from genes encoding (i) the wild-type (wt) minor coat protein (cp) iii of the filamentous bacteriophage m13 of escherichia coli, (ii) a truncated version of cpiii (amino acid (aa) positions 198-406), (iii) the wt major cpviii, or (iv) a hybrid of interleukin-1 beta (il-1 beta; aa 10-152) and wt cpviii. recombinant (re-) phage obtained by phag ... | 1995 | 7698668 |
| features of the dna fingerprinting probe pitz1. | stringently controlled plasmids generate dna fingerprint patterns in mammals when used at low hybridization temperatures. in order to develop a probe for use in paternity testing in cattle we screened a bovine, partial genomic plasmid library with the pcr-amplified ori region of plasmid p1. of eight isolated clones one generated strong band patterns at high stringency in various mammalian species (data not shown). sequence analysis revealed an imperfect, compound dinucleotide repeat region, whic ... | 1995 | 7702209 |
| thermodynamic genetics of the folding of the b1 immunoglobulin-binding domain from streptococcal protein g. | a method has been developed to select proteins that are thermodynamically destabilized yet still folded and functional. the dna encoding the b1 igg-binding domain from group g streptococcus (strp g) has been fused to gene iii of bacteriophage m13. the resulting fusion protein is displayed on the surface of the phage thus enabling the phage to bind to igg molecules. in addition, these phage exhibit a small plaque phenotype that is reversed by mutations that destabilize the strp g domain. by selec ... | 1995 | 7716165 |
| analysis of viral dna, protein and envelope damage after methylene blue, phthalocyanine derivative or merocyanine 540 photosensitization. | although numerous photosensitizers have been used experimentally to decontaminate viruses in cellular blood components, little is known about their mechanisms of photoinactivation. using m13 bacteriophage and vesicular stomatitis virus (vsv) as model viruses, we have investigated alteration of the viral genome, protein and envelope after phototreatment. methylene blue (mb) and aluminum phthalocyanine tetrasulfonate (alpcs4) phototreatment inactivated bacteriophage m13 and decreased the fraction ... | 1995 | 7740085 |
| 2d exchange 31p nmr spectroscopy of bacteriophage m13 and tobacco mosaic virus. | two-dimensional (2d) exchange 31p nuclear magnetic resonance spectroscopy is used to study the slow overall motion of the rod-shaped viruses m13 and tobacco mosaic virus in concentrated gels. even for short mixing times, observed diagonal spectra differ remarkably from projection spectra and one-dimensional spectra. our model readily explains this to be a consequence of the t2e anisotropy caused by slow overall rotation of the viruses about their length axis. 2d exchange spectra recorded for 30% ... | 1995 | 7756532 |
| bacteriophage surface display of an immunoglobulin-binding domain of staphylococcus aureus protein a. | as a model system for the optimization of separation ligands by bacteriophage surface display, we have constructed a phage surface expression system for a single immunoglobulin-binding domain of protein a of staphylococcus aureus. protein a domain b is genetically fused to the gpiii adsorption protein of the filamentous bacteriophage m13, and hence displayed on the phage surface. phage displaying the protein a domain are selectively retained on human igg-sepharose. retention is due to specific p ... | 1994 | 7764430 |
| identification of the core residues of the epitope of a monoclonal antibody raised against glycoprotein d of herpes simplex virus type 1 by screening of a random peptide library. | random peptide libraries (rpl) displayed on the surface of a filamentous bacteriophage can be used to identify peptide ligands that interact with target molecules. we have used a 15-amino acid residue rpl displayed on bacteriophage m13 to identify the core residues within the epitope of a monoclonal antibody (mab) a16 which interacts with a continuous epitope restricted to amino acid residues 9 to 19 in the n-terminal region of glycoprotein d of herpes simplex virus type 1 (gd-1). the single pep ... | 1994 | 7805747 |
| an engineered disulfide bridge in the transmembrane region of phage m13 coat protein stabilizes the alpha-helical dimer. | a single cys-residue (cys24) was introduced into the 50-amino acid major coat protein of m13 bacteriophage as part of a two-site substitution (y24c-v31a) within the effective transmembrane (tm) segment (tyr21 to ile39) of the coat protein. mutant y24c-v31a was able to complete the phage life cycle and was shown to contain free sulfhydryls in the intact virus, as evidenced by susceptibility of y24c-v31a phage to alkylation by cys-specific 14c-iodoacetamide (14c-ian). in contrast, the protein solu ... | 1995 | 7818525 |
| construction and characterization of m13 bacteriophages displaying functional igg-binding domains of staphylococcal protein a. | staphylococcal protein a (spa) is ranked as a versatile probe in immunoassays because of its immunoglobulin g (igg)-binding capability. however, poor binding of spa to the igg of some laboratory animals and its inability to bind human igg3 restricts its universal utility. in the present study, dna encoding the four igg-binding domains of spa (e, d, a and b) or the b domain alone has been fused, in separate phagemid vectors, to the 5' end of gene 111 of the phage m13. upon infection by helper pha ... | 1994 | 7828906 |
| unusual splice sites in the e1a-e1b cotranscripts synthesized in adenovirus type 40-infected a549 cells. | the adenovirus e1 dna region consists of two transcription units, e1a and e1b. in this paper we report that the e1a-e1b cotranscripts containing sequences of both the e1a and e1b regions are synthesized during adenovirus type 40 (ad40) infection of a549 cells. cytoplasmic rna was isolated from ad40-infected a549 cells at 24, 72, and 100 h post infection (p.i.). the complementary (c) dna was synthesized by reverse transcription using an oligo-dt primer and then amplified by the polymerase chain r ... | 1994 | 7832644 |
| histones associated with single-stranded dna do not preclude the formation of double-helical dna. | the effect of histones on the reaction of reassociation of the two complementary strands of dna from different sources has been investigated. the reassociation rate of denatured linear dna from bacteriophage m13 monitored spectrophotometrically and using nuclease s1 is roughly the same in the presence and absence of core histones at physiological ionic strength. electron microscopy reveals that in the samples containing histones a large network of duplex dna is produced. nevertheless, closed cir ... | 1995 | 7841189 |
| substitution of charged residues into the hydrophobic core of escherichia coli thioredoxin results in a change in heat capacity of the native protein. | two site-directed mutants of escherichia coli thioredoxin (l78k and l78r) were designed to study the effect of placing a charged residue in the hydrophobic core of the protein. both mutants retain catalytic activity in the assembly of phage m13. thermal denaturation of both these mutant proteins at ph 7.0 shows a reduction of stability of approximately 4 kcal.mol-1 with respect to the oxidized wild-type form. the thermal denaturation of the protein fits a dimeric state model. a significant reduc ... | 1995 | 7857925 |
| monoclonal antibodies against a minor and the major coat proteins of filamentous phage m13: their application in phage display. | we have produced monoclonal antibodies (mabs) which react with a minor and the major coat proteins of filamentous phage m13 and have characterised them by combining the techniques of enzyme-linked immunosorbent assay (elisa) and western blotting. these coat proteins are the minor coat protein, giiip, the product of gene iii and the major coat protein, gviiip, the product of gene viii. both giiip and gviiip are important in the context of 'phage display' of foreign peptides/proteins as fusions to ... | 1995 | 7876566 |
| [analysis of dna polymorphism detected by genomic fingerprinting based on phage m13 dna, in populations of bashkir and komi]. | hyperpolymorphism of minisatellite dna, detected using the m13 bacteriophage dna hybridization probe, was studied in three ethnographic groups of bashkirs and in the komi population. bands from 2 to 20 kb were analyzed in hybridization patterns. a significant population difference was detected both in evaluation of the average number of hybridization fragments per individuals and in the distribution of frequency of some fractions. thus, it seems possible to describe a set of so-called characteri ... | 1994 | 7890151 |
| fusion expression vectors for recombinant gene products processed easily and purified rapidly by affinity chromatography. | a dna fragment encoding igg-binding domain b,c (pabc) was separated from protein a gene, cloned into phage m13 and modified by oligo-directed mutagenesis at the hydroxylamine-cleaved site from asn-gly to asn-ala in domain b and c, respectively. the modified pabcm gene fragment was used to construct one set of fusion expression vectors in different reading frames. processing sequences such as those recognized by enterkinase, collagenase, thrombin, activated factor x and cleaved by the hydroxylami ... | 1994 | 7893935 |
| escherichia coli f plasmid transfers to and replicates within legionella pneumophila: an alternative to using an rp4-based system for gene delivery. | derivatives of the self-transmissible f plasmid of escherichia coli can be introduced into legionella pneumophila by conjugation and maintained within only upon selection. in l. pneumophila. f-based replicons seem to exist as extrachromosomal elements since they were readily lost when f-containing l. pneumophila was grown on nonselective medium. the f-based plasmids were not self-transmissible in l. pneumophila. the mating defect may be due to an inability to form the f pilus since f-containing ... | 1994 | 7899513 |
| comparative study of mutagenesis by o6-methylguanine in the human ha-ras oncogene in e. coli and in vitro. | single residues of o6-methylguanine (o6-meg) were introduced into the first or second position of codon 12 (ggc; positions 12g1 or 12g2, respectively) or the first position of codon 13 (ggt; position 13g1) of the human ha-ras oncogene in phage m13-based vectors. after transformation of e.coli, higher mutant plaque frequencies (mpf) were observed at 12g1 and 13g1 than at 12g2 if o6-alkylguanine-dna alkyltransferase (agt) had been depleted, while similar mpf were observed at all three positions in ... | 1994 | 7937103 |
| location of m13 coat protein in sodium dodecyl sulfate micelles as determined by nmr. | the major coat protein (gviiip) of bacteriophage m13 solubilized in sodium dodecyl sulfate (sds) detergent micelles was used as a model system to study this protein in the lipid-bound form. in order to probe the position of gviiip relative to the sds micelles, stearate was added, spin-labeled at the 5- or 16-position with a doxyl group containing a stable nitroxide radical. the average position of the spin-labels in the micelles was derived from the line broadening of the resonances in the 13c s ... | 1994 | 7947703 |
| fingerprinting reveals gamma-ray induced mutations in fungal dna: implications for identification of patent strains of trichoderma harzianum. | we have analyzed different patent strains and gamma-ray induced mutants of trichoderma harzianum by dna fingerprinting and pcr fingerprinting (rapd). applying wild-type phage m13 dna, with the oligonucleotides (ct)8 and (gtg)5 as probes for hybridization, as well as the oligonucleotides ggcatcggcc, (gtg)5, (cac)5 and the m13 sequence gagggtggcggttct as primers in pcr, we were able to obtain different and discriminative fingerprint patterns for all strains and mutants investigated. irradiation of ... | 1994 | 7954899 |
| display of expression products of cdna libraries on phage surfaces. a versatile screening system for selective isolation of genes by specific gene-product/ligand interaction. | techniques for cloning cdnas from bacteriophage libraries immobilised on solid supports are well established. however, these techniques do not allow selective enrichment of clones expressing proteins of interest. screening of cdna libraries would be simplified if the proteins encoded by cdnas could be expressed on the surface of phage. phage carrying genes encoding proteins for which a ligand is available can be selected directly by affinity interaction [crameri, r. & suter, m. (1993) gene (amst ... | 1994 | 7957259 |
| [a new method of covalent immobilization of oligodeoxyribonucleotides on nylon membranes for hybridization with nucleic acids]. | a new method of covalent immobilization of oligodeoxyribonucleotides on nylon membranes which contain surface amino groups was developed. the method consists in condensation between the amino group of the membrane and the carboxyl group of modified oligonucleotide by means of 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide. the carboxyl group was introduced into the oligonucleotide by means of postsynthetic attachment of peptide (reduced glutathione) at the terminal phosphate group of the oligonu ... | 1994 | 7990840 |