Publications
| Title | Abstract | Year Filter | PMID(sorted descending) Filter |
|---|
| replication of bacteriophage m13 dna in plasmolysed escherichia coli cells. | 1973 | 4585689 | |
| synthesis of phage m13 specific proteins in a dna-dependent cell-free system. | 1973 | 4584628 | |
| transmission of lac by the sex factor e in erwinia strains from human clinical sources. | lactose-utilizing (lac(+)) strains of erwinia spp. from human clinical material transfer lac by conjugation to plant strains of erwinia herbicola and erwinia amylovora, to other erwinia strains from human clinical sources, and also to escherichia coli, paracolobactrum arizonae, salmonella typhimurium, and shigella dysenteriae. the frequency of this transfer varies with the donor and recipient strains employed. the lac genes appear stable in these exconjugants, and they are not cured by acridine ... | 1973 | 4582635 |
| replication of bacteriophage m13. vii. requirement of the gene 2 protein for the accumulation of a specific rfii species. | 1972 | 4567400 | |
| a coat protein of the bacteriophage m13 virion participates in membrane-oriented synthesis of dna. | several molecules of a protein specified by gene 3 of m13 comprise a minor fraction of the phage coat and have been assigned a role in adsorption to the bacterial cell. we find that the gene-3 protein molecules of the virion are fully conserved in phage that have attached irreversibly to the host cell, and they form a complex with the phage dna when it has been converted to a duplex replicative form. in cells infected at a restrictive temperature with a thermosensitive mutant in gene 3, there is ... | 1973 | 4567335 |
| transfer among erwinia spp. and other enterobacteria of antibiotic resistance carried on r factors. | antibiotic resistance carried on r factors was transferred by conjugation from escherichia coli b/r and shigella flexneri 1a to erwinia spp. tetracycline resistance (tetr) carried on r factor r100 drd-56 was transferred from e. coli b/r to strains of erwinia amylovora, e. aroideae, e. atroseptica, e. chrysanthemi, e. cytolytica, e. dissolvens, e. herbicola, e. nigrifluens, and e. nimipressuralis, but not to strains of erwinia carotovora, e. carnegieana, e. dieffenbachiae, e. oleraceae, and e. qu ... | 1972 | 4562410 |
| replication of bacteriophage m13: specificity of the escherichia coli dnab function for replication of double-stranded m13 dna. | infection of the temperature-sensitive e. coli mutant hfrh 165/70 (dnab) with the filamentous single-stranded dna phage m13 is abortive at the restrictive temperature. upon infection at 41 degrees , single-stranded phage dna penetrates the cell and is converted in a rifampicin-sensitive step to the double-stranded replicative form (rf). the parental rf attaches to the cell membrane, but subsequent replication of the rf is blocked. it is concluded that in m13 infection semiconservative rf replica ... | 1972 | 4560691 |
| nucleotide sequencing of dna: preliminary characterization of the products of specific cleavages at guanine, cytosine, or adenine residues (bacteriophage m13-ribosubstitution-dna polymerase i-electrophoresis-two-dimensional fingerprinting). | dna synthesized in vitro from a phage m13 template has been cleaved at either guanine, adenine, or cytosine residues by ribosubstitution techniques. fingerprints of the fragments obtained suggest that dna sequencing will be possible with this technique. | 1972 | 4500550 |
| proceedings: regulation of gene activity in bacteriophage m13 dna. | 1974 | 4461536 | |
| bacteriophage m13 dna-directed in vitro synthesis of gene 5 protein. | 1974 | 4412163 | |
| isolation of mutants in m13 coat protein that affect its synthesis, processing, and assembly into phage. | the major coat protein (gene 8 protein) of bacteriophage m13 has been studied intensively as a model of membrane assembly, protein packing, and protein-dna interactions. because this protein is essential for assembly of the phage, very few mutants have been isolated. we have therefore cloned the gene 8 into a plasmid under control of the arab promoter. in the presence of arabinose, the cloned gene is expressed at a rate comparable to that in an m13-infected cell. plasmid-derived procoat is inser ... | 1985 | 4066698 |
| genes 55, alpha gt, 47 and 46 of bacteriophage t4: the genomic organization as deduced by sequence analysis. | the nucleotide sequence of t4 genes 55, alpha gt, 47 and 46 was determined by a combination of 'classical' procedures and a shotgun approach. small dna fragments generated by frequent cleavage with restriction enzymes or by sonication of restriction fragments were cloned in phage m13 vectors and sequenced by the dideoxy method. the positions of the genes were determined by marker rescue between the corresponding t4 amber mutants and the cloned t4 dna fragments used in the sequencing experiments. ... | 1985 | 4018026 |
| high-level expression of m13 gene ii protein from an inducible polycistronic messenger rna. | bacteriophage m13 gene ii has been cloned in the plasmid expression vector ping1 and thereby placed under the control of the inducible arab promoter of salmonella typhimurium. upon induction with arabinose, gene ii is transcribed as part of a polycistronic messenger rna which initiates at the arab promoter. subsequent translation of this message results in the coordinate, high-level expression of several proteins, including the gene ii protein. using this expression system, we have been able to ... | 1985 | 4007491 |
| 3-methylcholanthrene-induced expression of the cytochrome p-450c gene. | transcriptional control of 3-methylcholanthrene-dependent cytochrome p-450c nuclear rna induction was directly observed in an in vitro rat liver nuclear transcription system. mercurated and radiolabeled ribonucleotides were incorporated into nuclear rna transcribed in vitro, which was then isolated using thiopropyl-sepharose 6b affinity chromatography. dot hybridization experiments were carried out using bacteriophage m13 subclones of prsa57 (a cdna clone for rat serum albumin), peb339 (a cdna c ... | 1985 | 4004253 |
| supercoil sequencing: a fast and simple method for sequencing plasmid dna. | a method for obtaining sequence information directly from plasmid dna is presented. the procedure involves the rapid preparation of clean supercoiled plasmid dna from small bacterial cultures, its complete denaturation by alkali, and sequence determination using oligodeoxyribonucleotide-primed enzymatic dna synthesis in the presence of dideoxynucleoside triphosphates. the advantages of the method include speed, simplicity, avoidance of additional cloning steps into single-stranded phage m13 vect ... | 1985 | 3996185 |
| mutational mechanisms by which an inactive replication origin of bacteriophage m13 is turned on are similar to mechanisms of activation of ras proto-oncogenes. | m13 viral strand synthesis is initiated by nicking of the viral strand of the duplex replicative form by the m13 gene ii initiator protein at a specific site within a sequence of about 40 base pairs having dyad symmetry. efficient replication of the m13 viral strand also requires the presence of an adjacent sequence of ca. 100 base pairs. together these sequences constitute the minimal origin for m13 viral strand synthesis. a pbr322 derivative having a 182-base-pair insert of m13 dna contains a ... | 1985 | 3973968 |
| potential z-dna-forming elements in serum dna from human systemic lupus erythematosus. | dna fragments were isolated from serum of a patient with systemic lupus erythematosus. the majority of the dna was between 150 and 250 base pairs in length. the dna was cloned into phage m13, and 10 recombinants were sequenced. the average gc content of the dna was higher than total human dna (43% against 38%), with some fragments as high as 63%. this dna is rich in alternating purine-pyrimidine segments that are potentially z-dna-forming regions. | 1985 | 3964812 |
| [specific modification of dna at e. coli rna-polymerase binding sites]. | specific modification of promoter regions of dna has been studied. plasmid pk56b1 dna has been used as a model to test rna-polymerase binding with dna under various conditions. rna-polymerase is shown to form specific complexes with dna which are stable in solutions with a moderate ionic strength (0.1-0.2 m nacl), under ph 5-8 in the presence of 0.5 m o-methylhydroxylamine of o-delta-aminooxybutylhydroxylamine. escherichia coli jm103 cells have been transfected with dnas treated with 0.5 m o-met ... | 1985 | 3916215 |
| conserved residues of the leader peptide are essential for cleavage by leader peptidase. | gene 8 of bacteriophage m13 codes for procoat, the precursor of its major coat protein. gene 8 has been cloned into a plasmid and mutagenized. we have isolated mutants of this gene in which procoat is synthesized but is not processed to coat protein. we now describe mutants in the leader region of procoat, at positions -6, -3, and -1 with respect to the leader peptidase cleavage site. these positions are quite conserved among the leader peptides of various pre-proteins. each of these mutant proc ... | 1985 | 3905798 |
| partial purification of an enzyme from saccharomyces cerevisiae that cleaves holliday junctions. | an enzyme from saccharomyces cerevisiae that cleaves holliday junctions was partially purified approximately 500- to 1000-fold by deae-cellulose chromatography, gel filtration on sephacryl s300, and chromatography on single-stranded dna-cellulose. the partially purified enzyme did not have any detectable nuclease activity when tested with single-stranded or double-stranded bacteriophage t7 substrate dna and did not have detectable endonuclease activity when tested with bacteriophage m13 viral dn ... | 1985 | 3903750 |
| m13 procoat inserts into liposomes in the absence of other membrane proteins. | procoat, the precursor form of the major coat protein of coliphage m13, assembles into the escherichia coli inner membrane and is cleaved to mature coat protein by leader peptidase. this assembly process has previously been reconstituted using lipids and purified leader peptidase in a cell-free protein synthesis reaction (watts, c., silver, p., and wickner, w. (1981) cell 25, 347-353; ohno-iwashita, y., and wickner, w. (1983) j. biol. chem. 258, 1895-1900). we now report that procoat can also cr ... | 1985 | 3902814 |
| sequence-dependent termination of in vitro dna synthesis by cis- and trans-diamminedichloroplatinum (ii). | inhibition of dna replication by the antitumor drug cis-diamminedichloroplatinum (ii) (cis-ddp) has been proposed to be responsible for its cytotoxicity. treatment of primed phage m13 mp8 viral dna templates with the drug followed by second-strand synthesis using large fragment dna polymerase i reveals that cis-ddp forms an adduct with dna that inhibits dna synthesis in vitro. this inhibition occurs at all (dg)n (n greater than or equal to 2) sequences in the template strand, confirming that the ... | 1985 | 3895221 |
| membrane protein conformational change dependent on the hydrophobic environment. | two conformational states of the coat protein of the filamentous bacteriophage m13 have been detected in detergent solution by using magnetic resonance techniques. when 3-fluorotyrosine is incorporated in place of the two tyrosine residues in the protein, four 19f nuclear magnetic resonance signals are observed, two for each conformer of the protein. the equilibrium between the two forms can be modulated by ph, temperature, and detergent structure. the rate of interconversion of the isomers is r ... | 1985 | 3893541 |
| a new strategy to create ordered deletions for rapid nucleotide sequencing. | a method is described for generating ordered deletions using previously published techniques but a new strategy. this method is simpler than the published ones and has many advantages. target dna is cloned in both orientations into one of the unique restriction enzyme sites adjacent to the complementary region of the commercially available primers in bacteriophage m13. ordered unidirectional deletions are created using bal 31 nuclease and religating into m13 vector dna without the need of purify ... | 1985 | 3891520 |
| base-pairing properties of n4-methoxydeoxycytidine 5'-triphosphate during dna synthesis on natural templates, catalyzed by dna polymerase i of escherichia coli. | n4-methoxydeoxycytidine 5'-triphosphate (mo4dctp) was synthesized by reaction of dctp with methoxyamine and then purified by high-performance liquid chromatography (hplc) and used to analyze the specificity of mo4dcmp incorporation during polymerization on natural templates, catalyzed by dna polymerase i of escherichia coli. elongation of synthetic 5'-32p-labeled primers, annealed to single-stranded dna of bacteriophage m13, was carried out in the presence of only three of the four normal dntps; ... | 1985 | 3888268 |
| folding of single-stranded dna on the histone octamer. | a complex between the single-stranded dna of the bacteriophage m13 and the histone octamer was analyzed by electron microscopy, low-angle x-ray diffraction and nuclease analysis. the morphology and the diffraction pattern of the complex strongly resemble those of the nucleosome. these results, as well as the finding of a protected dna fragment about 100 nucleotides long following single-stranded dna specific nuclease digestion, indicate that 'a nucleosome-like' complex can be formed between sing ... | 1985 | 3882454 |
| cloning, nucleotide sequence, and overexpression of the bacteriophage t4 rega gene. | the bacteriophage t4 rega gene codes for a regulatory protein that controls the expression of a number of t4 early genes, apparently at the level of translation. restriction fragments containing the rega structural gene have been cloned into phage m13, and the nucleotide sequence has been determined. translation of the dna sequence predicted that rega protein contains 122 amino acids, with a mr of 14,620. a dna fragment carrying 85% of the coding sequence of rega has been cloned into the phage l ... | 1985 | 3872458 |
| [isolation of mutant genes for human leukocyte alpha2-interferon by a method of localized mutagenesis]. | an efficient method to obtain the mutant genes for human leucocyte alpha 2-interferon (ifn) has been elaborated. the technique includes the following main stages: cloning of interferon gene in m13mp8 dna; isolation of double-stranded hybrid dna complex, containing ifn gene as a single-stranded fragment; selective modification of a single-stranded hybrid dna by sodium bisulphite; the repair of hybrid dna by dna polymerase i from escherichia coli, transformation of escherichia coli jn103 cells by ... | 1985 | 3842756 |
| the region of phage t4 genes 34, 33 and 59: primary structures and organization on the genome. | the product of gene 33 is essential for the regulation of late transcription and gene product 59 is required in recombination, dna repair and replication. the exact functions of both proteins are not known. restriction fragments spanning the genomic area of genes 33 and 59 have been cloned into phage m13 and a 4.9 kb nucleotide sequence has been determined. translation of the dna sequence predicted that gp33 contains 112 amino acids with a mol.wt. of 12.816 kd while gp59 is composed of 217 amino ... | 1986 | 3797242 |
| biosynthesis of reovirus-specified polypeptides. molecular cdna cloning and nucleotide sequence of the reovirus serotype 1 lang strain s3 mrna which encodes the nonstructural rna-binding protein sigma ns. | human reovirus serotype 1 lang strain s3 mrna, which encodes the nonstructural rna-binding polypeptide sigma ns, was cloned as a cdna:mrna heteroduplex in escherichia coli using phage m13. a complete consensus nucleotide sequence was determined. the lang strain s3 mrna is 1198 nucleotides in length and possesses an open reading frame with a coding capacity of 366 amino acids, sufficient to account for a sigma ns polypeptide of 41,179 daltons. comparison of the serotype 1 (lang) s3 sequence with ... | 1986 | 3767989 |
| rapid preparation of bacteriophage dna for sequence analysis in sets of 96 clones, using filtration. | a method is described for the preparation of single-stranded dna from clones in bacteriophage m13 vectors. this procedure allows multiples of 96 clones to be processed at once, utilizing filtration to remove host cells and simplifying the treatment of bacteriophage pellets. the dna produced can be used for sequencing of mutagenesis. | 1986 | 3766942 |
| sequences complementary to the brain-specific "identifier" sequences exist in l-type pyruvate kinase mrna (a liver-specific messenger) and in transcripts especially abundant in muscle. | a sequence complementary to the brain-specific identifier sequence has been found in the 3' untranslated extension of the heavy 3.2-kilobase (kb) long liver l-type pyruvate kinase mrna while it is absent in the other two 2- and 2.2-kb long pyruvate kinase mrna species. a 53-base fragment corresponding to this identifier sequence was subcloned in both orientations in the single-stranded bacteriophage m13, both strands being used as probes to detect homologous sequence in different tissues. both s ... | 1986 | 3753703 |
| nucleotide sequence identity of mitochondrial dna from different human tissues. | recombinant dna techniques have been used to search for mitochondrial (mt) nucleotide (nt) sequence differences between human tissues within an individual. mtdna isolated from brain, heart, liver, kidney, and skeletal muscle of two different individuals was cleaved with saci and xbai, and then cloned in bacteriophage m13. partial nt sequence determination of 121 independently isolated recombinant m13 clones containing either the cytochrome oxidase subunit iii gene or the d-loop region of human m ... | 1986 | 3744049 |
| synthesis of a fixed-length single-stranded dna probe by blocking primer extension in bacteriophage m13. | a simple and efficient technique has been developed for preparing radiolabeled single-stranded (ss) probes of determined length and high specific radioactivity. the human beta-globin gene intervening segment ii (ivsii) fragment (0.9-kb) was inserted between the ecori and bamhi sites of m13mp11 and used as a template for ss probe synthesis. the m13 hybridization probe primer (m13 hpp) was annealed to the recombinant m13mp11-beta ivsii template dna. this m13 hpp was next blocked by the enzymatic a ... | 1986 | 3721200 |
| biosynthesis of reovirus-specified polypeptides. molecular cdna cloning and nucleotide sequence of the reovirus serotype 1 lang strain s2 mrna which encodes the virion core polypeptide sigma 2. | human reovirus serotype 1 lang strain s2 mrna, which encodes the virion inner capsid core polypeptide sigma 2, was cloned as a cdna:mrna heteroduplex in escherichia coli using phage m13. a complete consensus nucleotide sequence was determined. the lang strain s2 mrna is 1331 nucleotides in length and possesses an open reading frame with a coding capacity of 335 amino acids, sufficient to account for a sigma 2 polypeptide of 37,682 daltons. comparison of the serotype 1 lang s2 sequence derived fr ... | 1987 | 3663211 |
| biosynthesis of reovirus-specified polypeptides. efficiency of expression of cdnas of the reovirus s1 and s4 genes in transfected animal cells differs at the level of translation. | full-length cdnas of the reovirus serotype 1 lang strain s1 and s4 genes were cloned in escherichia coli using bacteriophage m13 and expressed in monkey cos cells under the control of the sv40 late promoter using the eukaryotic expression vector pjc119. the s1-encoded sigma 1 and s4-encoded sigma 3 gene products were expressed in transfected cos cells and were indistinguishable from the authentic sigma 1 and sigma 3 polypeptides synthesized in reovirion-infected cos cells. the relative translati ... | 1987 | 3617502 |
| primary structure of an alpha-tubulin gene of physarum polycephalum. | an alpha-tubulin gene of physarum was isolated as a phage-lambda nm1149 recombinant (designated phage-lambda n alpha tu). phage-lambda n alpha tu contained a 4700 base-pair hindiii nuclear dna fragment of an allele of the altb locus of physarum (one of four unlinked alpha-tubulin gene loci). subfragments of the 4700 base-pair insert of phage-lambda n alpha tu were cloned into phage m13 and the nucleotide sequence was determined by the dideoxy chain termination method. the start point of transcri ... | 1987 | 3586027 |
| screening recombinant clones containing sequences homologous to escherichia coli genes using single-stranded bacteriophage vector. | detection and isolation of escherichia coli clones carrying vectors with foreign dna sequences partially homologous to specific e. coli genes is difficult because denatured dna in the host genome can hybridize with the probe. in this paper we present a procedure which simplifies this task by using bacteriophage m13 as the cloning vector. the procedure takes advantage of the secretory properties of the phage, as well as the property of nitrocellulose membrane to bind protein and single-stranded d ... | 1986 | 3549464 |
| a sensitive colorimetric detection of virus dna and oncogene. | advantage of cloning probe dna fragment in phage m13 dna was taken to provide a larger single stranded dna as a hybridization probe. high level of direct enzyme labels was introduced via the m13 dna moiety as well as probe dna. a highly sensitive colorimetric detection of virus dna and oncogene was developed. | 1987 | 3548725 |
| structure and expression of the alkb gene of escherichia coli related to the repair of alkylated dna. | when the alkb gene of escherichia coli that controls sensitivity of bacteria to methyl methanesulfonate was placed under the control of the lac regulatory region on a multicopy plasmid, the gene product, alkb protein, was overproduced. by monitoring the band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein was purified to near physical homogeneity. an amino-terminal sequence and total amino acid composition of the purified alkb protein were in accord with the amino acid ... | 1986 | 3536913 |
| selection by genetic transformation of a saccharomyces cerevisiae mutant defective for the nuclear uracil-dna-glycosylase. | a coliphage m13 chimer containing the saccharomyces cerevisiae trp1 gene and ars1 replication origin (mpy2) was grown on an ung- dut- strain of escherichia coli. the resulting single-stranded phage dna had 13% of thymine residues substituted by uracil. this dna failed to transform a delta trp1 yeast strain to prototrophy. however, when a mutagenized yeast stock was transformed with uracil-containing single-stranded mpy2 dna, unstable transformants were obtained. after plasmid segregation, about ... | 1986 | 3519585 |
| biosynthesis of reovirus-specified polypeptides. molecular cdna cloning and nucleotide sequence of the reovirus serotype 1 lang strain s4 mrna which encodes the major capsid surface polypeptide sigma 3. | serotype 1 lang strain s4 mrna, which encodes the major capsid surface polypeptide sigma 3 of reovirions, was cloned as a cdna:mrna heteroduplex in escherichia coli using phage m13. a complete consensus nucleotide sequence for s4 mrna has been determined from cdna clones. the lang strain s4 mrna is 1196 nucleotides in length and possesses an open reading frame with a coding capacity of 365 amino acids, sufficient to account for a sigma 3 polypeptide of 41,212 daltons. comparison of the serotype ... | 1986 | 3518713 |
| backbone dynamics of a model membrane protein: 13c nmr spectroscopy of alanine methyl groups in detergent-solubilized m13 coat protein. | the filamentous coliphage m13 possesses multiple copies of a 50-residue coat protein which is inserted into the inner membrane of escherichia coli during infection. 13c nuclear magnetic resonance (nmr) spectroscopy has been used to probe the structure and dynamics of m13 coat protein solubilized in detergent micelles. a comparison of backbone dynamics within the hydrophobic core region and the hydrophilic terminal domains was obtained by biosynthetic incorporation of [3-13c]alanine. alanine is d ... | 1986 | 3513830 |
| tobacco mosaic virus replicase and replicative structures. | the rna-dependent rna polymerase (replicase) mediating the replication of tobacco mosaic virus (tmv) has been investigated in a number of laboratories over a period of 20 years. cell-free enzyme preparations have been prepared which can continue the synthesis of nascent complementary rna, initiated in vivo; however, the enzyme does not require, nor does it respond to, exogenous viral rna as a template. the presence in plants of a virus-stimulated, host-encoded rna-dependent rna polymerase (rdrp) ... | 1987 | 3503886 |
| different dna-binding modes and cooperativities for bacteriophage m13 gene-5 protein revealed by means of fluorescence depolarisation studies. | the binding of the bacteriophage-m13-encoded gene-5 protein to oligo(deoxythymidylic acid)s and m13 dna was studied by means of tyrosyl fluorescence decay and fluorescence anisotropy measurements. the observed fluorescence decays could be described with two exponentials, characterised by the lifetimes tau 1 = 2.2 ns and tau 2 = 0.8 ns respectively. only the amplitude of the longer-lifetime component is influenced by binding of the protein to dna. this indicates that a part of the tyrosyl residue ... | 1986 | 3486763 |
| isolation of lactoferrin cdna from a human myeloid library and expression of mrna during normal and leukemic myelopoiesis. | lactoferrin is a major constituent of polymorphonuclear leukocyte granules and is present in mature neutrophils but not in blasts or promyelocytes. we have isolated a cdna probe for lactoferrin and used it to study the synthesis of lactoferrin mrna by normal and leukemic granulocyte precursors. the probe phl-41 has been subcloned in phage m13 and characterized by restriction endonuclease analysis and nucleic acid sequencing. phl-41 contains approximately 40% of the coding sequence of the lactofe ... | 1987 | 3477300 |
| [cloning of dna complementary to mrna for proopiomelanocortin from the bovine, rat and human hypophysis. hormonal regulation of proopiomelanocortin mrna in the rat hypophysis]. | cloning of dna and complementary mrna of bovine, rat and human proopiomelanocortin (pomc) was carried out. a structural analysis of the cloned cdna of pomc was performed. using restriction fragments of bovine, rat and human pomc cloned cdna, probes for molecular hybridization based on one-chain bacteriophage m13 were made. using the dot-hybridization technique with labeled [32p] pomc cdna, the effect of 17 beta-estradiol and adrenalectomy on the pomc mrna level in rat hypophysis was studied. the ... | 1987 | 3474031 |
| human delta-aminolevulinate dehydratase: nucleotide sequence of a full-length cdna clone. | two cdnas encoding human delta-aminolevulinate dehydratase (ala-d; porphobilinogen synthase; ec 4.2.1.24), the second enzyme in the heme biosynthetic pathway, were identified, recloned into bacteriophage m13, and sequenced by primer extension. the first clone with an 827-base-pair (bp) pex-ala-d cdna insert, shown to contain dna sequences that were colinear with four bovine ala-d peptide sequences, was used to screen a pkt218 human liver library. a second clone containing a 1200-bp insert was id ... | 1986 | 3463993 |
| 5-azido-2'-deoxyuridine 5'-triphosphate: a photoaffinity-labeling reagent and tool for the enzymatic synthesis of photoactive dna. | we have synthesized the photoactive deoxyuridine nucleotide 5-azido-2'-deoxyuridine 5'-triphosphate (5-n3dutp) and used it to synthesize light-sensitive dna by enzymatic incorporation. in the absence of ultraviolet light, 5-n3dutp is a substrate for escherichia coli dna polymerase i. in in vitro dna synthesis reactions using bacteriophage m13 single-stranded dna as the template and 5-n3dutp in place of dttp, a photoactive complementary strand was synthesized by dna polymerase i. the complementar ... | 1986 | 3461438 |
| oligonucleotide-directed construction of mutations: a gapped duplex dna procedure without enzymatic reactions in vitro. | the gapped duplex dna approach to oligonucleotide-directed construction of mutations (kramer et al. 1984, nucl. acids res. 12, 9441-9456) has been developed further. a procedure is described that makes in vitro dna polymerase/dna ligase reactions dispensable. direct transfection of host bacteria with gddna molecules of recombinant phage m13 plus mutagenic oligonucleotide results in marker yields in excess of 50% (gap size 1640 nucleotides). an important feature incorporated into the mutagenic ol ... | 1988 | 3405755 |
| calf thymus dna polymerases alpha and delta are capable of highly processive dna synthesis. | we have demonstrated that calf thymus dna polymerases alpha and delta are capable of highly processive dna synthesis. processivity values between 300 and 2000 nucleotides were observed when poly(da)-oligo(dt) or singly primed single-stranded circular bacteriophage m13 dna at ph 6.0 and 1 mm magnesium chloride was used. these conditions do not correlate with conditions, ph 7.0 and 5 mm magnesium chloride, that support the maximum synthetic rate. lowering the ph and magnesium concentration lowers ... | 1988 | 3401462 |
| rabbit liver factor d, a poly(thymidine) template stimulatory protein of dna polymerases: purification and characterization. | factor d, a dna binding protein that enhances the activities of diverse dna polymerases with a common restricted set of templates, was initially characterized in mouse liver but has resisted extensive purification. in this paper, we report that a similar stimulatory activity can be obtained in highly purified form from nuclei of rabbit hepatocytes. the rabbit liver protein increases the rates at which several dna polymerases copy sparsely primed natural dna templates and primed synthetic poly(dt ... | 1988 | 3401461 |
| characteristics of lactose-fermenting salmonella strains from poland. | in this study 184 lactose-fermenting salmonella strains, collected in the national salmonella centre from the northern and central parts of ponad were examined. epidemiological, serological and biochemical investigations were carried out. apart from this, chemotherapeutic resistance and male-phage sensitivity were determined. most of strains belonged to s. agona serotype (s. typhimurium and s. oranienburg were also presented) which apart from the lactose-fermenting ability retained all the remai ... | 1987 | 3333475 |
| [design of a hybrid gene coding for the leader sequence of bacillus amyloliquefaciens alpha-amylase and for human proinsulin]. | the chemically synthesized structure gene of human proinsulin was cloned in e. coli on the secretory vector containing regulatory elements of the bacillus amyloliquefaciens alpha-amylase gene. the proinsulin gene was inserted by the ecori site located immediately after the dna area encoding the alpha-amylase signal peptide. the e. coli cells transformed by such a plasmid produced hybrid protein consisting of the alpha-amylase signal peptide, five amino acid residues after the gene mating and hum ... | 1987 | 3326519 |
| [formation and properties of artificial polycistrons containing truncated genes for e. coli tryptophan operon and phage m13 envelope protein]. | using gene fragments encoding the leader peptide of e. coli tryptophane operon (as duplicated fragment hhai-140) or m13 phage coat protein (as taqi-381 or haeiii-1623 fragments) and basing on pds1 family of plasmids, expression vectors have been constructed which contained transcription promoters ptrp, pviii, and pv + pviii, respectively. an artificial gene for human leukocyte interferon alpha 2 (ifn-alpha 2) has been cloned into these plasmids, so that its transcription was a part of polycistro ... | 1987 | 3322290 |
| time-resolved tryptophan fluorescence anisotropy investigation of bacteriophage m13 coat protein in micelles and mixed bilayers. | coat protein of bacteriophage m13 is examined in micelles and vesicles by time-resolved tryptophan fluorescence and anisotropy decay measurements and circular dichroism experiments. circular dichroism indicates that the coat protein has alpha-helix (60%) and beta-structure (28%) in 700 mm sodium dodecyl sulfate micelles and predominantly beta-structure (94%) in mixed dimyristoylphosphatidylcholine/dimyristoylphosphatidic acid (80/20 w/w) small unilamellar vesicles. the fluorescence decay at 344 ... | 1987 | 3318926 |
| bacteriophage m13 procoat protein inserts into the plasma membrane as a loop structure. | the major coat protein of bacteriophage m13 is synthesized as a precursor, the procoat, with a typical leader (signal) sequence of 23 residues at its nh2-terminus. a fusion protein that contains the nh2-terminal 141 residues of cytoplasmic ribulokinase and all but the first ten residues of m13 procoat was made. the fusion protein inserts into the plasma membrane of escherichia coli and is processed by leader peptidase to give rise to a leader peptide of 155 residues and the mature coat protein o ... | 1987 | 3317833 |
| solution hybridization of crosslinkable dna oligonucleotides to bacteriophage m13 dna. effect of secondary structure on hybridization kinetics and equilibria. | several dna oligonucleotides have been photochemically modified with the furocoumarin 4'-hydroxymethyl-4,5',8-trimethylpsoralen (hmt) such that each contained a single hmt furan side monoadduct to thymidine at a unique 5' tpa 3' sequence. when these oligonucleotides were hybridized to their respective complements, the hmt adduct could be driven to form an interstrand crosslink by irradiation of the hybrid with 360 nm light. the ability to crosslink probe-target complexes has allowed us to determ ... | 1987 | 3316669 |
| deoxyhexanucleotide containing a vinyl chloride induced dna lesion, 1,n6-ethenoadenine: synthesis, physical characterization, and incorporation into a duplex bacteriophage m13 genome as part of an amber codon. | organic synthesis and recombinant dna techniques have been used to situate a single 1,n6-ethenoadenine (epsilon ade) dna adduct at an amber codon in the genome of an m13mp19 phage derivative. the deoxyhexanucleotide d[gct(epsilon a)gc] was chemically synthesized by the phosphotriester method. mild nonaqueous conditions were employed for deprotection because of the unstable nature of the epsilon ade adduct in aqueous basic milieu. physical studies involving fluorescence, circular dichroism, and 1 ... | 1987 | 3314993 |
| backbone dynamics of a model membrane protein: assignment of the carbonyl carbon 13c nmr resonances in detergent-solubilized m13 coat protein. | the major coat protein of the filamentous bacteriophage m13 is a 50-residue amphiphilic polypeptide which is inserted, as an integral membrane-spanning protein, in the inner membrane of the escherichia coli host during infection. 13c was incorporated biosynthetically into a total of 23 of the peptide carbonyls using labeled amino acids (alanine, glycine, lysine, phenylalanine, and proline). the structure and dynamics of carbonyl-labeled m13 coat protein were monitored by 13c nuclear magnetic res ... | 1987 | 3307912 |
| homologous recombination intermediates between two duplex dna catalysed by human cell extracts. | using as substrates, 1: the replicative form (rf) of phage m13 mp8 in which the reading frame of the lac z' gene was disrupted by insertion of an octonucleotide, and 2: a restriction fragment one kb long, containing the functional lac z' gene (isolated from wild type m13 mp8), we show that nuclear extracts from human cells (3 lines tested) promote the targeted replacement of the altered sequence by the functional one. following incubation with the extracts, the dna's were introduced in jm 109 ba ... | 1987 | 3302944 |
| inhibition of purified escherichia coli leader peptidase by the leader (signal) peptide of bacteriophage m13 procoat. | the leader peptide of bacteriophage m13 procoat inhibited the cleavage of m13 procoat or pre-maltose-binding protein by purified escherichia coli leader peptidase. this finding confirms inferences that the leader is the primary site of enzyme recognition and suggests a rationale for the rapid hydrolysis of leader peptides in vivo. | 1987 | 3301818 |
| high frequencies of short frameshifts in poly-ca/tg tandem repeats borne by bacteriophage m13 in escherichia coli k-12. | slipped-strand mispairing (ssm) may play an major role in repetitive dna sequence evolution by generating large numbers of short frameshift mutations within simple tandem repeats. here we examine the frequency and size spectrum of frameshifts generated within poly-ca/tg sequences inserted into bacteriophage m13 in escherichia coli hosts. the frequency of detectable frameshifts within a 40 bp tract of poly-ca/tg is greater than one percent and increases more than linearly with length, being lower ... | 1987 | 3299269 |
| the dna sequence specificity of stimulation of dna polymerases by factor d. | the mechanism of enhancement of dna polymerase activity by the murine dna-binding protein factor d was investigated. extension by escherichia coli dna polymerase i and calf thymus dna polymerase-alpha of 5'-32p-labeled oligodeoxynucleotide primers that are complementary to poly(dt) or to bacteriophage m13 dna was measured in the absence or presence of factor d. with 5'-[32p](da)9.poly(dt), factor d enables e. coli polymerase i to fill approximately 15-nucleotide gaps between adjacent primers; wh ... | 1987 | 3298245 |
| sequence comparison of single-stranded dna binding proteins and its structural implications. | the primary sequences were compared among several proteins: gene product 5 protein (gp5) from phage m13; pike from phage ike; gene product 32 protein (gp32) from phage t4; reca, ssb and ssf from escherichia coli. these proteins bind strongly and cooperatively to single-stranded dna with no sequence specificity. gp5 is the smallest in this group and its three-dimensional structure is well-characterized. using the entire sequence of gp5 as a template we searched for the regions in other single-str ... | 1987 | 3295261 |
| effects of the escherichia coli ssb protein on the binding of escherichia coli reca protein to single-stranded dna. demonstration of competitive binding and the lack of a specific protein-protein interaction. | the effect of the escherichia coli single-stranded dna binding (ssb) protein on the stability of complexes of e. coli reca protein with single-stranded dna has been investigated through direct dna binding experiments. the effect of each protein on the binding of the other to single-stranded dna, and the effect of ssb protein on the transfer rate of reca protein from one single-stranded dna molecule to another, were studied. the binding of ssb protein and reca protein to single-stranded phage m13 ... | 1987 | 3295259 |
| mutant 16s ribosomal rna: a codon-specific translational suppressor. | we have isolated an unusual codon-specific translational suppressor in escherichia coli. the suppressor resulted from a spontaneous mutation in a chromosomal gene during a selection for suppressors of the auxotrophic nonsense mutation trpa(uga211). the suppressor allows readthrough of uga mutations at two positions in trpa and at two sites in bacteriophage t4. it does not, however, suppress amber (uag) or ochre (uaa) mutations that were tested in both genomes, some of which were at the same posi ... | 1988 | 3288986 |
| detection of toxigenic escherichia coli using biotin-labelled dna probes following enzymatic amplification of the heat labile toxin gene. | several types of dna probes labelled with biotin were compared for their sensitivity to detect the heat labile toxin (lt) gene in toxigenic escherichia coli. in addition, a procedure was developed for enzymatically amplifying lt gene sequences in toxigenic e. coli. probes were labelled with biotinylated nucleotides by either nick translation; 3' tailing; primer extension of probe dna cloned into bacteriophage m13; sandwich hybridization; or oligolabelling of isolated dna fragments. a single stra ... | 1988 | 3288864 |
| [genomic fingerprints of organisms from different taxonomic groups: the use of phage m13 dna as a hybridization probe]. | hypervariable polymorphic patterns were detected using wild-type m13 dna as a probe in genomic dnas of very different organisms ranging from procaryotes and lower eucaryotes to upper plants and animals, including human beings. due to somatic stability of highly polymorphic patterns and their discrete inheritance, individual-specific restriction pattern analysis ("dna fingerprinting") with this test probe was found to be useful in applied human genetics, in particular, for identifying paternity a ... | 1988 | 3282990 |
| sequence-specific chemical modification of a hybrid bacteriophage m13 single-stranded dna by alkylating oligonucleotide derivatives. | alkylating oligonucleotide derivatives react with the complementary sequences in hybrid m13mp7 bacteriophage single-stranded dna and destroy the infecting ability of the dna. the reagents do not damage m13mp9 single-stranded dna lacking the target nucleotide sequence. | 1988 | 3282926 |
| dna-binding properties of gene-5 protein encoded by bacteriophage m13. 2. further characterization of the different binding modes for poly- and oligodeoxynucleic acids. | the binding of gene-5 protein, encoded by bacteriophage m13, to oligodeoxynucleic acids was studied by means of fluorescence binding experiments, fluorescence depolarization measurements and irreversible dissociation kinetics of the protein.nucleotide complexes with salt. the binding properties thus obtained are compared with those of the binding to polynucleotides, especially at very low salt concentration. it appears that the binding to oligonucleotides is always characterized by a stoichiomet ... | 1988 | 3262511 |
| sequence analysis of a processed gene coding for mouse ribosomal protein l32. | the nucleotide sequence of a mouse ribosomal protein gene, identified by hybridization with the gene encoding the drosophila ribosomal (r-) protein 49, was determined by cloning in the phage m13 and dideoxy sequencing. the mouse gene, l32', is a member of the multigene family encoding mammalian r-protein l32. l32' is a processed gene that could encode a 135 amino acid protein similar to that of mouse l32 and drosophila r-protein 49. | 1988 | 3246356 |
| cloning and determination of the nucleotide sequence of the mn-containing superoxide dismutase gene from halobacterium halobium. | a group of synthetic 17-mer oligodeoxynucleotides (oligos) was constructed to correspond to a sequence of amino acids situated near the n terminus of the manganese-containing superoxide dismutase (mn-sod) purified from the halophilic bacterium, halobacterium halobium. a cosmid library of a sau3ai partial digest of halobium dna, cloned into the bamhi site of phc79, was probed with the radiolabeled oligos. cosmid dna was purified from the clone that showed hybridization at the highest stringency. ... | 1988 | 3240866 |
| [effectiveness of distal gene translation in polycistrons depends upon the arrangement of regulatory signals on a template]. | the role of the translational terminator and initiator signals arrangement for two adjacent genes in polycistronic mrna has been studied. semisynthetic beta-galactosidase gene (lacz) of e. coli and fragment of phage m13 dna (with promoter pviii, gene ix, and part of gene viii) were used for constructing of the ix-viii-lacz artificial polycistronic operon. cloning of the constructs into pbr322 vector resulted in a number of plz381n plasmids differing by the mutual arrangement of gene viii transla ... | 1988 | 3233097 |
| [complementary-addressed elimination of e1a sequence of simian adenovirus oncogene sa7 from circular single-stranded dna of recombinant phage m13]. | the g fragment of the simian adenovirus sa7 oncogene corresponding to e1a region was cloned into m13mp8 and m13mp9 phages. single-stranded dnas of the recombinant phages thus obtained (mp8g and mp9g) partially digested with dnase ii were used to synthesize polyalkylating derivatives capable of specific hybridisation and subsequent alkylation of complementary g sequences of corresponding phage dnas. after incubation of complementary alkylated dna in the presence of lysine, the preselected region ... | 1988 | 3219135 |
| characterization of a sequence of human t cell leukemia virus type i from a patient with chronic progressive myelopathy. | dna from peripheral blood mononuclear cells of individuals with chronic progressive myelopathy (cpm) were extensively analyzed for the presence of human t cell leukemia virus (htlv) type i-like sequences by using the polymerase chain reaction. the dna samples were amplified with oligonucleotides from three separate regions of htlv viral genomes. a portion of the amplified viral genome from a representative patient was sequenced after molecular cloning into bacteriophage m13. sequence data indica ... | 1988 | 3198935 |
| [complementary addressed modification of single- and double-stranded dna by alkylating derivatives of oligonucleotides isolated by partial dna fragmentation]. | reagents for complementary addressed modification of nucleic acids are proposed to be synthesized on the base of oligonucleotides obtained by partial chemical fragmentation of dna. the alkylating 4-(n-2 chlorethyl-n-methylamino) benzyl-5'-phosphamide derivatives of 5'-[32p]-labelled oligonucleotides obtained from single and double-stranded dna cloned in bacteriophage m13 mp9 have been synthesized. the alkylated derivatives of oligonucleotides selectively modify the complementary tracts of single ... | 1988 | 3193969 |
| deuterium nuclear magnetic resonance investigation of bacteriophage m13 coat protein in dimyristoylphosphatidylcholine liposomes using palmitic acid as a probe. | the effect of incorporation of various amounts of m13 bacteriophage coat protein on the bilayer order and acyl chain motion in dimyristoylphosphatidylcholine (dmpc) liposomes has been investigated using deuterium nmr of specifically deuterated palmitic acid as a bilayer probe, phosphorus nmr and additional spin-label electron spin resonance (esr). the secondary structure of the m13 coat protein in these bilayers was determined from circular dichroism spectra. phosphorus nmr spectra of the mixed ... | 1988 | 3179303 |
| chemical and biological studies of the major dna adduct of cis-diamminedichloroplatinum(ii), cis-[pt(nh3)2(d(gpg]], built into a specific site in a viral genome. | a duplex escherichia coli bacteriophage m13 genome was constructed containing a single cis-[pt(nh3)2(d(gpg]] intrastrand cross-link, the major dna adduct of the anticancer drug cis-diamminedichloroplatinum(ii). the duplex dodecamer d(agaaggcctaga).d(tctaggccttct) was ligated into the hincii site of m13mp18 to produce an insertion mutant containing a unique stui restriction enzyme cleavage site. a genome with a 12-base gap in the minus strand was created by hybridizing hincii-linearized m13mp18 d ... | 1988 | 3166983 |
| repetitive dna sequences as probes for mycobacterium tuberculosis. | three cloned segments of mycobacterium tuberculosis dna which are promising as clinical probes were identified. an mboi digest of dna from a clinical isolate of m. tuberculosis was cloned into bacteriophage m13. to identify recombinants specific for the m. tuberculosis complex, plaque lifts were hybridized with m. bovis and m. kansasii dna. recombinants which selectively hybridized with m. bovis dna were characterized by probing slot blots and restriction digests of dna from various mycobacteria ... | 1988 | 3148630 |
| effects of sos and mucab functions on reactivation and mutagenesis of m13 replicative form dna bearing bulky lesions. | we have previously determined the specificity of -1 frameshifts induced by aflatoxin-b1-2,3-dichloride (afb1c12) in phage m13 double-strand replicative form (rf) dna. the system consists of: (i) in vitro adduction of rf dna of bk8, a lacz + 1 frameshift derivative of phage m13mp8; (ii) transfection into unirradiated or uv-irradiated bacterial host cells; (iii) scoring and sequencing of revertants (i.e., -1 frameshifts). the previous data had shown that induction of sos functions enhanced mutagen ... | 1988 | 3141805 |
| preferential uptake of restriction fragments from a gonococcal cryptic plasmid by competent neisseria gonorrhoeae. | factors involved in the specificity of dna uptake by competent neisseria gonorrhoeae were examined. host-controlled modification did not affect uptake. certain restriction fragments of the 4.2 kb gonococcal cryptic plasmid pfa1 and of the replicative form of the bacteriophage m13 were taken up in preference to others, independent of differences in fragment size. a 600 bp fragment from the 4.2 kb plasmid was cloned into ples2, a gonococcal-escherichia coli shuttle vector; the 600 bp fragment was ... | 1988 | 3141569 |
| cloning and sequence analysis of cytadhesin p1 gene from mycoplasma pneumoniae. | mycoplasma pneumoniae cytadhesin p1 was purified by monoclonal antibody affinity chromatography followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. the n-terminal 18-amino-acid sequence of p1 was determined and used to design two synthetic oligonucleotides, a 14-mer corresponding to amino acids 1 to 5 and an 18-mer corresponding to amino acids 7 to 12. these oligonucleotides served as hybridization probes for the identification of the p1 gene by southern blot analy ... | 1987 | 3119495 |
| purified scrapie prions resist inactivation by procedures that hydrolyze, modify, or shear nucleic acids. | prions were purified from scrapie-infected hamster brains and incubated for 24 hr at 65 degrees with 2 mm zn2+ or 5 mm mg2+; no loss of infectivity was observed. bacteriophage m13, tobacco mosaic virus (tmv), potato virus x, and potato spindle tuber viroid were all inactivated by divalent metal ions under these conditions. prions also resisted inactivation by prolonged digestions with dnase i, rnases a and t1, and micrococcal nuclease. prions were resistant to psoralen photoadduct formation usin ... | 1987 | 3114950 |
| purified scrapie prions resist inactivation by uv irradiation. | the development of effective purification protocols has permitted evaluation of the resistance of isolated scrapie prions to inactivation by uv irradiation at 254 nm. prions were irradiated on ice with doses of uv light ranging up to 120,000 j/m2. uv dosimetry experiments, performed with saccharomyces cerevisiae plasmid dna or eucaryotic cells, indicated that under these experimental conditions an incident uv dose of 10 j/m2 formed 2 thymine dimers per 5.1 x 10(6) daltons of eucaryotic cell dna. ... | 1987 | 3097336 |
| specificity of ionizing radiation-induced mutagenesis in the lac region of single-stranded phage m13 mp10 dna. | m13 mp10 single-stranded phage dna was irradiated with 60 co gamma-rays, and transfected into escherichia coli. one hundred and sixteen mutant clones having lesions in the lac insert were selected, and mutational sites were examined by dna sequence analysis. fourteen out of the 15 nucleotide changes thus detected were base substitutions, and the rest was a base addition. transitions and transversions were almost equal in number. mutational events were observed at cytosine residues more frequentl ... | 1986 | 3088546 |
| secretion and export of igf-1 in escherichia coli strain jm101. | the processing of lamb-igf-1 fusion protein and the export of processed igf-1 (insulin-like growth-factor-1) into the growth medium was examined in the escherichia coli host strain, jm101. several strain or plasmid modifications were tried to increase export of periplasmic (processed) igf-1 into the growth medium of jm101. these included: (1) use of a lon null mutant strain to increase accumulation levels of unprocessed lamb-igf-1 fusion protein; (2) use of an alternative drug resistance marker ... | 1988 | 3071740 |
| [expression in escherichia coli of dna coding for human tumor necrosis factor]. | the variants of expression in escherichia coli of artificial dna coding for human tumor necrosis factor, an important immune modulator with selective cytotoxic action on a number of transformed cell lines have been described. the dna was placed under control of either phage m13 promoter of gene for main coat protein or tandem of pair of e. coli tryptophane promoters. it has been shown that e. coli cells harbouring plasmids described with artificial tnf gene provide good level of protein biosynth ... | 1988 | 3071369 |
| [a simple system of cloning in phage m13 and dna sequencing with terminators]. | a compilation of techniques for dna cloning in filamentous phage m13 based vectors for a novice in cloning is presented. it does not require either specialized microbiological facilities, or any specific knowledge in escherichia coli genetics. the cloning strategy uses only blunt-end ligation into a vector that has been prepared once for several hundred experiments. the first part describes the isolation, preparation and checking of a blunt-ended m13 vector (with m13 mp series vectors as an exam ... | 1988 | 3065614 |
| a novel replicon occurring naturally in escherichia coli is a phage-plasmid hybrid. | a novel dna replicon in escherichia coli was identified. it is the smallest natural isolate (1282 bp) found so far. in the presence of phage m13 it grows as a filamentous single-stranded dna phage. contrary to previously identified mini-phages this replicon displays sequence homology only to parts of the m13 viral and complementary strand origin. in the absence of m13 this dna replicates autonomously. the only gene (arp) of the replicon encodes a 32-kd protein, which is essential for autonomous ... | 1988 | 3061812 |
| [genomic fingerprinting of microorganisms: its use as a hybridization probe of phage m13 dna]. | hypervariable nucleotide sequences detected by hybridization with the phage m13 dna probe were found in the chromosomal dnas of certain pathogenic microbial species. dna fingerprinting, based on hybridization of m13-probe with hypervariable chromosomal dna sequences, opens new approaches to epidemiological analysis, epidemiological prognosis, taxonomy, and other theoretical and applied fields of bacteriology. | 1988 | 3053332 |
| nucleotide sequence analysis and overexpression of the gene encoding a type iii chloramphenicol acetyltransferase. | the gene catiii, encoding a type iii enterobacterial chloramphenicol acetyltransferase, was cloned from the transmissible plasmid r387 into pbr322 and bacteriophage m13 mp8. nucleotide sequence analysis of 1160 bp of dna identified an open reading frame encoding a protein of 213 amino acid residues and a calculated molecular mass of 24965 da. the predicted n-terminal sequence is identical with that determined by edman degradation of chloramphenicol acetyltransferase purified from escherichia col ... | 1988 | 3048245 |
| structure and dynamics of a detergent-solubilized membrane protein: measurement of amide hydrogen exchange rates in m13 coat protein by 1h nmr spectroscopy. | the coat protein of bacteriophage m13 is inserted into the inner membrane of escherichia coli where it exists as an integral membrane protein during the reproductive cycle of the phage. the protein sequence consists of a highly hydrophobic 19-residue central segment flanked by an acidic 20-residue n-terminus and a basic 11-residue c-terminus. we have measured backbone amide hydrogen exchange of the protein solubilized in perdeuteriated sodium dodecyl sulfate using 1h nuclear magnetic resonance ( ... | 1988 | 3042022 |
| effects of temperature-sensitive variants of the bacillus subtilis dnab gene on the replication of a low-copy-number plasmid. | the dnab gene of bacillus subtilis is involved in the initiation of dna replication and also in the binding of the chromosomal origin to the bacterial membrane. we studied the effect of temperature-sensitive dnab mutants (dnab1 and dnab19) on the replication and on the dna-membrane binding of the plasmid pkw1, which was derived from the low-copy-number plasmid pbs2. in the dnab19 mutant, pkw1 was not able to replicate at the restrictive temperature. in the dnab1 mutant, however, the dimeric form ... | 1987 | 3040678 |
| dna mismatch-repair in escherichia coli counteracting the hydrolytic deamination of 5-methyl-cytosine residues. | derivatives of phage m13 were constructed and used for the in vitro preparation of heteroduplex dna molecules containing base/base mismatches that mimick dna lesions caused by hydrolytic deamination of 5-mec residues in escherichia coli dna (i.e. they carry a t/g mismatch in the special sequence context provided by the recognition site -cca/tgg-of the dcm-methyltransferase). upon introduction of these heteroduplex dnas into cacl2-treated e. coli cells, the mismatches are efficiently repaired wit ... | 1987 | 3038536 |
| recombinant forms of m13 procoat with an ompa leader sequence or a large carboxy-terminal extension retain their independence of secy function. | the assembly of phage m13 procoat protein into the plasma membrane of escherichia coli is independent of the secy protein. to test whether this is caused by the unusually small size of procoat, we fused dna encoding 103 amino acids to the carboxy-terminal end of the procoat gene. the resulting fusion protein, which attains the same membrane-spanning conformation as mature coat protein, still does not require the secy function for membrane assembly. to determine whether the leader sequence govern ... | 1987 | 3034592 |
| differential gene expression during the amoebal-plasmodial transition in physarum. | we have prepared cdna libraries for amoebae and plasmodia of the acellular slime mould, physarum polycephalum. differential screening was used to isolate cell-type-specific cdna clones (in bacteriophage m13) and both libraries yielded approximately 5% of such sequences. the amoebal- and plasmodial-specific clones were used to assay changes in transcription during the amoebal-plasmodial transition. the results obtained substantiate the view that the switch from amoebal to plasmodial characteristi ... | 1987 | 3029710 |
| determination of size and orientation of dna fragments cloned in phage m13 by s1 nuclease mapping. | 1987 | 3029677 | |
| [variants of phage m13 dna containing a fragment of the beta-galactosidase gene--a convenient mutation system for the study of oligonucleotide-directed mutagenesis]. | a model system is developed to test oligonucleotide-directed mutations: t----c transition, t and c deletions (delta t and delta c), c insertion, double mutations (a----g, delta t), (t----c, a----g), and large oligonucleotide deletions (36 or 44 nucleotides). the system includes 9 variants of the phage m13 dna carrying fragment of beta-galactosidase gene, and oligodeoxyribonucleotides partially noncomplementary to dna sequence of this gene. six variants are obtained by the site-localized mutagene ... | 1986 | 3028430 |
| coliphage m13 cloning system and ddxtp chain-termination method for dna sequencing. | two dna fragments of cytochrome b gene in yeast mitochondrial dna were sequenced by messing's m13 cloning system and sanger's ddxtp chain-termination method. m13mp8 and m13mp9 serve as vectors for insert fragment. the recipient strain is e. coli jm103 for transfection. when the ratio of insert/vector was 3:1, high frequencies of recombination and positive recombination were obtained. the two fragments, which have 575 bp and 709 bp, were sequenced. to read more bases, the ratio of ddxtp/dxtp must ... | 1986 | 3027889 |