Publications

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
the nucleotide sequence of the akv murine leukemia virus genome.	the nucleotide sequence of an infectious molecular clone of the akv murine leukemia virus has been determined by the dideoxy chain termination method after subcloning in bacteriophage m13 vectors. the sequence predicts an rna genome of 8371 nucleotides containing three large open reading frames corresponding to the gag, pol, and env genes. signal sequences for transcription, splicing, and translation have been identified. the positions of 95 major rnase t1 resistant oligonucleotides of the akv r ...	1984	6200992
cloning and structure determination of cdna for cutinase, an enzyme involved in fungal penetration of plants.	the primary structure of cutinase, an extracellular fungal enzyme involved in the penetration of plants by pathogenic fungi, has been determined from the nucleotide sequence of cloned cdna. clones containing cdna made from poly(a)(+) rna isolated from fungal cultures induced to synthesize cutinase were screened for their ability to hybridize with the [(32)p]cdna for mrna unique to the induced culture. the 75 cdna clones thus identified were screened for the cutinase genetic code by hybrid-select ...	1984	16593482
subgenomic rnas with nucleotide sequences derived from rnas 1 and 2 of cucumber mosaic virus can act as messenger rnas in vitro.	encapsidated rnas of cucumber mosaic virus (cmv) were analyzed by hybridization to specific probes after gel electrophoresis. [32p]-complementary dna (cdna) probes were prepared by transcription of genomic rna 1 and rna 2 nucleotide sequences that had been cloned in a bacteriophage m13 vector. probes that correspond to unique sequences near the 3' ends of rna 1 and rna 2 revealed over 20 smaller rnas. the subgenomic rnas derived from each genomic rna were analyzed more definitively by hybrid sel ...	1985	18639845
on the origin of the helper component of tobacco vein mottling virus: translational initiation near the 5' terminus of the viral rna and termination by uag codons.	the nature of the polypeptide products encoded by the 5'-terminal region of the rna of the potyvirus, tobacco vein mottling virus (tvmv), was investigated. single-stranded dna probes complementary to either nucleotides 1100-2100 or 2100-2820 from the 5' terminus of the rna were prepared by subcloning recombinant plasmids in bacteriophage m13. these were hybridized to tvmv rna, the dna:rna hybrids translated in a reticulocyte lysate cell-free translation system (hybrid-arrested translation), and ...	1985	18639848
replication of tobacco mosaic virus. viii. characterization of a third subgenomic tmv rna.	in an earlier study we concluded that tobacco mosaic virus (tmv) infections engender a third subgenomic rna in infected tissue (p. palukaitis, f. garcia-arenal, m. a. sulzinski, and m. zaitlin (1983), virology 131, 533-545). this rna of approximate mw of 1.1 x 10(6), termed i1-rna, was shown to be polyribosome-associated and thus was presumed to serve as a messenger rna in vivo. upon in vitro translation of i1-rna in a rabbit reticulocyte lysate system, a major product of mw approximately 50k wa ...	1985	18640547
lysogenisation of spiroplasma citri by a type 3 spiroplasmavirus.	a short-tailed polyhedral spiroplasmavirus, ai, with a plaque morphology typical of temperate phages and a linear double-stranded dna genome which can circularise due to the presence of cohesive ends, was able to lysogenise spiroplasma citri. lysogenic sporoplasmas spontaneously released ai virus at a low level, and were immune to superinfection by the released virus, but not to infection by two serologically related viruses. these properties were retained following repeated cultivation in virus ...	1985	18640554
[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
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
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
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
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
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
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
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
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
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
[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
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
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
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
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
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
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
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
initiation of dna replication on single-stranded dna templates catalyzed by purified replication proteins of bacteriophage lambda and escherichia coli.	initiation of bacteriophage lambda dna replication at the chromosomal origin depends on the lambda o and p replication proteins. these two viral initiators, together with an escherichia coli protein fraction, promote the replication in vitro of single-stranded circular dna chromosomes such as that of bacteriophage m13. this nonspecific strand initiation reaction, which we have termed the "lambda single-strand replication reaction," has now been established with eight purified proteins, each of w ...	1985	2408273
specificity of the binding of bacteriophage m13 encoded gene-5 protein to dna and rna studied by means of fluorescence titrations.	the fluorescence quenching of the bacteriophage m13 encoded gene-5 protein was used to study its binding characteristics to different polynucleotides. experiments were performed at different salt concentrations and in some instances at different temperatures. the affinity of the protein depends on the base and sugar composition of the polynucleotides involved and may differ appreciably, i.e. by orders of magnitude. the salt dependence of binding is within experimental accuracy equal for all sing ...	1985	2482044
dna primase-dna polymerase alpha from simian cells: sequence specificity of initiation sites on simian virus 40 dna.	unique single-stranded regions of simian virus 40 dna, phage m13 virion dna, and several homopolymers were used as templates for the synthesis of (p)pprna-dna chains by cv-1 cell dna primase-dna polymerase alpha. intact rna primers, specifically labeled with an rna capping enzyme, were typically 6 to 8 ribonucleotides long, although their lengths ranged from 1 to 9 bases. the fraction of intact rna primers 1 to 4 ribonucleotides long was 14 to 73%, depending on the template used. rna primer leng ...	1985	2582240
non-radioactive hybridization probes prepared by the chemical labelling of dna and rna with a novel reagent, photobiotin.	a photo-activatable analogue of biotin, n-(4-azido-2-nitrophenyl)-n'-(n-d-biotinyl-3-aminopropyl)-n'-methyl-1,3- propanediamine (photobiotin), has been synthesized and used for the rapid and reliable preparation of large amounts of stable, non-radioactive, biotin-labelled dna and rna hybridization probes. upon brief irradiation with visible light, photobiotin formed stable linkages with single- and double-stranded nucleic acids yielding probes which were purified from excess reagent by 2-butanol ...	1985	2582358
improved m13 phage cloning vectors and host strains: nucleotide sequences of the m13mp18 and puc19 vectors.	three kinds of improvements have been introduced into the m13-based cloning systems. (1) new escherichia coli host strains have been constructed for the e. coli bacteriophage m13 and the high-copy-number puc-plasmid cloning vectors. mutations introduced into these strains improve cloning of unmodified dna and of repetitive sequences. a new suppressorless strain facilitates the cloning of selected recombinants. (2) the complete nucleotide sequences of the m13mp and puc vectors have been compiled ...	1985	2985470
cloning and dna sequence of a plasmid-determined citrate utilization system in escherichia coli.	the citrate utilization determinant from a large 200-kilobase (kb) naturally occurring plasmid was previously cloned into the psti site of plasmid vector pbr325 creating the cit+ tetracycline resistance plasmid pwr61 (15 kb). tn5 insertion mutagenesis analysis of plasmid pwr61 limited the segment responsible for citrate utilization to a 4.8-kb region bordered by ecori and psti restriction nuclease sites. the 4.8-kb fragment was cloned into phage m13, and the dna sequence was determined by the di ...	1985	2999088
screening for thermostable mutant of kanamycin nucleotidyltransferase by the use of a transformation system for a thermophile, bacillus stearothermophilus.	a structural gene of kanamycin nucleotidyltransferase cloned into a single-stranded bacteriophage m13 was subjected to mutagenesis with hydroxylamine. having recloned the mutagenized gene of the enzyme in a vector plasmid ptb922, the recombinant plasmid was used to transform bacillus stearothermophilus with a purpose of screening for the more thermostable enzyme than the wild type. out of greater than 8 x 10(3) transformants, 12 clones that were suspected to harbor the mutant gene encoding the m ...	1985	2999110
a complete library of point substitution mutations in the glucocorticoid response element of mouse mammary tumor virus.	the glucocorticoid response element (gre) of mouse mammary tumor virus (mmtv) was chemically synthesized as two complementary dna strands bearing cohesive termini. during automated synthesis, random mutations were introduced into the dna by "doping" each of the four nucleoside phosphoramidites (a, g, c, and t) with a low level of the other three. these preparations were annealed and cloned into an m13 phage vector to produce a library of gre mutants. mutations within the synthesized region were ...	1986	3003746
illegitimate recombination at the replication origin of bacteriophage m13.	hybrids composed of phage m13 and plasmid phv33 were used to study the formation of deletions in escherichia coli. eighty to ninety percent of the deletion endpoints were at the position of the nick introduced into the m13 replication origin by the phage gene ii protein. this suggests the existence of a novel mechanism of illegitimate recombination.	1986	3010295
dna alterations photosensitized by tetracycline and some of its derivatives.	bacteriophage m13 mp10 dna were irradiated with near-uv light in the presence of tetracycline derivatives and primed with synthetic oligonucleotide to be used for dna synthesis using escherichia coli dna polymerase. chain terminations were observed by denaturing polyacrylamide gel electrophoresis and mapped precisely. all the synthesis stops occurred before or at the level of guanine residues, showing that the photoreaction mediated by tetracycline derivatives led to a preferential alteration of ...	1986	3011916
hydrolysis by restriction endonucleases at their dna recognition sequences substituted with mismatched base pairs.	restriction endonucleases were tested for their ability to catalyze the cleavage of mismatch-containing recognition sites in dna. these mismatched base pairs were t.g, u.g, or a.c in covalently closed, circular heteroduplexes prepared by in vitro extension of chemically synthesized oligonucleotide primers annealed to a bacteriophage m13-derived viral dna. none of the restriction enzymes was able to completely cleave the mismatch-containing recognition sites under standard conditions. however, th ...	1986	3012472
nucleotide sequence and deduced amino acid sequence of escherichia coli pyruvate oxidase, a lipid-activated flavoprotein.	the entire nucleotide sequence of the poxb (pyruvate oxidase) gene of escherichia coli k-12 has been determined by the dideoxynucleotide (sanger) sequencing of fragments of the gene cloned into a phage m13 vector. the gene is 1716 nucleotides in length and has an open reading frame which encodes a protein of mr 62,018. this open reading frame was shown to encode pyruvate oxidase by alignment of the amino acid sequences deduced for the amino and carboxy termini and several internal segments of th ...	1986	3016647
site-directed mutagenesis of the binding site for ribosomal protein s8 within 16s ribosomal rna from escherichia coli.	twelve specific alterations have been introduced into the binding site for ribosomal protein s8 in escherichia coli 16s rrna. appropriate rdna segments were first cloned into bacteriophage m13 vectors and subjected to bisulfite and oligonucleotide-directed mutagenesis in vitro. subsequently, the mutagenized sequences were placed within the rrnb operon of plasmid pno1301 and the mutant plasmids were used to transform e. coli recipients. the growth rates of cells containing the mutant plasmids wer ...	1986	3016664
tarantula hemocyanin mrna. in vitro translation, cdna cloning and nucleotide sequence corresponding to subunit e.	following induction of hemopoiesis, poly(a)-rich rna was prepared from the heart of the tarantula, eurypelma californicum, and translated in rabbit reticulocyte lysates. in vitro translation products were immunoprecipitated with antiserum against whole dissociated eurypelma hemocyanin. analysis of the immunoprecipitate by sodium dodecyl sulfate/polyacrylamide gel electrophoresis revealed a set of polypeptides comigrating with authentic eurypelma hemocyanin. the mrna was transcribed into cdna, cl ...	1986	3017715
nucleotide sequence of cdna and derived amino acid sequence of rabbit complement component c3 alpha-chain.	the nucleotide sequence coding for 726 amino acid residues of the alpha-chain of rabbit c3 was determined from a cdna clone. subfragments of the cdna produced by restriction endonucleases were inserted into the bacteriophage m13 and sequenced using the dideoxynucleotide technique. the derived amino acid sequence was compared with those of human and mouse c3, which have been previously reported [by de brujn, m.h.l. and fey, g.h. (1985) proc. natl. acad. sci. usa 82, 708, and westel, r.a. et al. ( ...	1986	3019881
nucleotide sequence of the structural genes for an anion pump. the plasmid-encoded arsenical resistance operon.	the structural genes for the arsenical pump of the conjugative r-factor r773 contained on a hindiii fragment of 4.3 kilobase pairs were cloned into bacteriophage m13. a series of ordered deletions was created using bal31 digestion, and the nucleotide sequence of the operon determined. three open reading frames for genes arsa, arsb, and arsc were found. the arsa gene encodes a hydrophilic protein of 63,169 da with two potential adenylate-binding sites. the arsb gene encodes a potentially membrane ...	1986	3021763
[insertion of the cos-site into dna of phage m13 and its packing in proteins of phage lambda].	the cos-site of lambda phage from phc79 cosmide is transferred to dna from m13 mp18 phage. the recombinant dna thus obtained (mc18) is efficiently packaged into lambda proteins in vitro. the bamhi-hindiii fragment of pgp588 (a pbr322 derivatives containing fragment of human dna) is subcloned into mc18. although this pgp588 fragment contains numerous alu repeats, no essential rearrangements of the insert were revealed. the efficiency infection by recombinant dna packaged with lambda proteins is a ...	1986	3022756
molecular characteristics of prion rods purified from scrapie-infected hamster brains.	purification of scrapie prions from hamster brains has demonstrated that the infectious particles contain one major protein, prp 27-30. this protein, which is required for and inseparable from scrapie infectivity, polymerizes into heterogeneous rod-shaped particles measuring 10-20 nm in diameter and 100-200 nm in length. we attempted to identify the minimal infectious unit by disrupting aggregates of the rods. prolonged sonication resulted in progressive fragmentation of the rods into spherical ...	1986	2872252
replication of uv-irradiated single-stranded dna by dna polymerase iii holoenzyme of escherichia coli: evidence for bypass of pyrimidine photodimers.	replication of uv-irradiated circular single-stranded phage m13 dna by escherichia coli rna polymerase (ec 2.7.7.6) and dna polymerase iii holoenzyme (ec 2.7.7.7) in the presence of single-stranded dna binding protein yielded full-length as well as partially replicated products. a similar result was obtained with phage g4 dna primed with e. coli dna primase, and phage phi x174 dna primed with a synthetic oligonucleotide. the fraction of full-length dna was several orders of magnitude higher than ...	1986	2941756
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
multiple crosslinks of proteins s7 and s9 to domains 3 and 4 of 16s ribosomal rna in the escherichia coli 30s particle.	rna-protein cross-links were introduced into escherichia coli 30s subunits by treatment with 1-ethyl-3(3-dimethylaminopropyl)carbodiimide. 16s rrna, cross-linked to 30s ribosomal proteins, was isolated and hybridized with seven single-stranded bacteriophage m13-dna probes. these probes, each carrying an inserted rdna fragment, were used to select contiguous rna sections covering domains 3 and 4 (starting at nucleotide 868 and ending at the 3'oh terminus) of the 16s rrna. the proteins covalently ...	1986	2429836
biosynthesis of reovirus-specified polypeptides. molecular cdna cloning and nucleotide sequence of the reovirus serotype 1 lang strain bicistronic s1 mrna which encodes the minor capsid polypeptide sigma 1a and the nonstructural polypeptide sigma 1bns.	human reovirus serotype 1 lang strain s1 mrna, which encodes the minor capsid cell attachment protein sigma 1a and the nonstructural protein sigma 1bns, was cloned as a cdna:mrna heteroduplex in escherichia coli using phage m13. the lang strain s1 mrna is 1462 nucleotides in length and possesses two open reading frames. the first begins at nt 14 and has a coding capacity of 418 amino acids, sufficient to account for sigma 1a; the second begins at nt 75 and has a coding capacity of 119 amino acid ...	1986	2430568
a simple and rapid nucleotide sequencing strategy and its application in analyzing a rice histone 3 gene.	an improved rapid method for sequencing a target dna is described. a new plasmid, paa-pz1, which contains the origin of replication from phage m13 and a portion of the tn9 transposon was constructed. a long fragment of target dna cloned into this vector is progressively shortened in vivo from one end by transposon-mediated deletions. the plasmids carrying different lengths of target dna are then made into single-stranded dna in the same host upon infection with an m13 phage and their sequence is ...	1986	3026911
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
[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
an analysis of tobacco mosaic virus replicative structures synthesized in vitro.	the rna structures synthesized in vitro by a crude enzyme complex from tobacco mosaic virus (tmv)-infected leaves have been analyzed; the major viral-specific products were similar to tmv-replicative form (rf) and-replicative intermediate (ri) in electrophoretic behavior and ribonuclease sensitivity. synthesis of these rf-like and ri-like structures neither required nor responded to added viral rna, but did require all four ribonucleotide triphosphates. enriched radiolabeled rf-like and ri-like ...	1986	24307422
determination of size and orientation of dna fragments cloned in phage m13 by s1 nuclease mapping.		1987	3029677
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
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
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
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
[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
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
genome interactions which influence dna palindrome mediated instability and inviability in escherichia coli.	the interaction of three factors determine the detrimental effect of a palindromic dna sequence in escherichia coli cells. the first is the nature of the palindrome (its length, extent of central asymmetry and perhaps its base sequence), the second is the genotype of the host cell and the third is the replicon within which it is located. in this paper we extend the genetic and physical characterization of lambda bacteriophages carrying a palindrome of approximately 560 base pairs. we also show t ...	1987	2972734
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
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
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
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
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
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
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
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
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
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
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
[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
[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
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
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
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
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
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
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
expression of a trna gene in the context of the lacz mrna.	fusions of the gene for tyrosine suppressor trna, tyrt(sup3), and the lacz gene of escherichia coli were constructed such that the trna gene could be expressed from either its own promoter or that of the lac operon. these chimeras, carried on phage m13 vectors, were tested for the expression of the trna in e. coli. the trna gene was expressed on the order of 10-fold more weakly from the lac promoter than from its own promoter. to examine whether pausing or premature termination of transcription ...	1987	3027034
analysis of the role of the cysteine 171 residue in the activity of herpes simplex virus type 1 thymidine kinase by oligonucleotide-directed mutagenesis.	the thymidine kinase (tk) gene from herpes simplex virus type 1 strain sc16 was cloned into bacteriophage m13 mp8 so that functional hsv-1 tk was expressed in bacteria infected with the recombinant bacteriophage, m13/tk. oligonucleotide site-directed mutagenesis was then employed to introduce single nucleotide changes into the tk gene in m13/tk in order to alter the codon for cysteine 171 in the wild-type enzyme to a codon specifying either serine or glycine. analysis of the mutant enzymes in ba ...	1987	3027247
a single base change in the shine-dalgarno region of 16s rrna of escherichia coli affects translation of many proteins.	a single base mutation was constructed at position 1538 of escherichia coli 16s rrna, changing a cytidine to a uridine. this position is in the shine-dalgarno region, thought to be involved in base-pairing to mrna during initiation of protein synthesis. the mutation was constructed by using a synthetic oligodeoxynucleotide that differs in sequence by one base from the wild-type sequence of 16s rrna. this oligonucleotide was used as a primer on single-stranded dna of phage m13, into which was clo ...	1987	2440027
a system for rapid dna sequencing with fluorescent chain-terminating dideoxynucleotides.	a dna sequencing system based on the use of a novel set of four chain-terminating dideoxynucleotides, each carrying a different chemically tuned succinylfluorescein dye distinguished by its fluorescent emission is described. avian myeloblastosis virus reverse transcriptase is used in a modified dideoxy dna sequencing protocol to produce a complete set of fluorescence-tagged fragments in one reaction mixture. these dna fragments are resolved by polyacrylamide gel electrophoresis in one sequencing ...	1987	2443975
effect of bacteriophage m13 infection on phosphorylation of dnak protein and other escherichia coli proteins.	1. the effects of infection with the filamentous phage m13 on the phosphorylation of escherichia coli proteins were studied. phosphorylated proteins were labeled with [32p]orthophosphate and analyzed by the o'farrell two-dimensional gel technique and autoradiography. 2. phage infection was shown to induce significant changes in the pattern of protein phosphorylation. at least eight different proteins were found to be phosphorylated to a larger extent while seven others were, by contrast, much le ...	1987	2822422