Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| clones from an 840-kb fragment containing the 5' region of the dmd locus enriched by pulsed field gel electrophoresis. | detailed analysis of a large region of genomic dna is facilitated by generating overlapping clones covering the entire region. these clones are usually obtained by bidirectional "walking" using either bacteriophage lambda or cosmid cloning vectors. this is a slow procedure when starting from a single start site. multiple start sites are an advantage, and here we describe a method of generating clones from an extensive region of the duchenne muscular dystrophy locus by preparative pulsed field ge ... | 1988 | 3066744 |
| recent data on the structure of rabbit milk protein genes and on the mechanism of the hormonal control of their expression. | mammary explants or isolated mammary cells from rabbit have been cultured in the presence of insulin, prolactin and cortisol alone or in combination. the cellular content in alpha s1-casein, beta-casein and whey acidic protein (wap) mrna have been evaluated using the corresponding cdna as probes. in all cases alpha s1-casein mrna was the most abundant and wap mrna the least abundant mrna. the three genes showed essentially similar dependency towards hormones. prolactin stimulated mrna accumulati ... | 1988 | 3072627 |
| molecular cloning and genetic analysis of the determinant for gamma-lysin, a two-component toxin of staphylococcus aureus. | the gamma-lysin determinant of staphylococcus aureus strain smith 5r has been cloned in phage lambda and plasmid vectors in escherichia coli. genetic evidence is presented which demonstrates that gamma-lysin requires the co-operative action of two polypeptides expressed by the closely linked hlga and hlgb genes. recombinants expressed haemolytic activity in agarose medium but not in agar, a known property of gamma-lysin. haemolysis was inhibited by antiserum raised against the 32 kda component o ... | 1988 | 3075655 |
| molecular biological search for human genes encoding cholinesterases. | cholinesterases (ches) are highly polymorphic proteins, capable of rapidly hydrolyzing the neurotransmitter acetylcholine and involved in terminating neurotransmission in neuromuscular junctions and cholinergic synapses. in an attempt to delineate the structure and detailed properties of the human protein(s) and the gene(s) coding for the acetylcholine hydrolyzing enzymes, a human cdna coding for che was isolated by use of oligodeoxynucleotide screening of cdna libraries. for this purpose, a met ... | 1987 | 3077058 |
| protein fusions of beta-galactosidase to the ferrichrome-iron receptor of escherichia coli k-12. | the fusion-generating phage lambda plac mu1 was used to produce fusions of lacz to fhua, the gene encoding the ferrichrome-iron receptor (fhua protein) in the outer membrane of escherichia coli k-12. fusions to the fhua gene in a delta (lac) strain were selected by their resistance to bacteriophage phi 80 vir. ten independent (fhua'-'lacz) fusions were all lac+ and were resistant to the lethal agents which require the fhua protein as receptor, i.e., phi 80 vir, t5, t1, uc-1, and colicin m; none ... | 1986 | 3079747 |
| inefficient translation initiation causes premature transcription termination in the lacz gene. | expression plasmids containing the e. coli lacz coding region preceded by a set of different ribosome-binding sites and put under transcriptional control of the leftward promoter of phage lambda (pl) were used to study the synthesis of lacz mrna. in a normal host the steady state level of full-length lacz mrna varied 100-fold with the different synthesis levels of beta-galactosidase, whereas in a host expressing the antitermination protein n of phage lambda, all vectors synthesized the same amou ... | 1986 | 3081264 |
| cloning and expression of the 3' portion of the t4 denv gene as a lacz fusion gene. | polyclonal antibodies have been raised against endonuclease v from the bacteriophage t4. this rabbit serum, from which endemic e. coli antibodies have been removed, reacts with a single protein from t4-infected e. coli with a molecular weight of 16078 dalton. it was confirmed that these antibodies were directed against endonuclease v through the inhibition of the pyrimidine dimer specific nicking activity of endonuclease v in an in vitro nicking assay. a phage lambda gt11 t4 dc dna library was s ... | 1986 | 3081797 |
| cell-type-specific and regulated expression of a human gamma 1 heavy-chain immunoglobulin gene in transgenic mice. | a functionally rearranged human gamma 1 heavy-chain immunoglobulin gene was cloned from a human plasma cell leukemia cell line, arh-77, into the phage lambda charon 4a. the recombinant phage dna was introduced into fertilized mouse eggs (about 200 copies of the human gene per egg). a total of 30 mice were born and were screened for the presence of the human gamma 1 gene by dot hybridization. two of these 30 mice had integrated one or two copies of the gene. the gamma 1 mrnas were detected only i ... | 1986 | 3083415 |
| expression of mycoplasma pneumoniae antigens in escherichia coli. | a genomic library of mycoplasma pneumoniae was generated by using bacteriophage lambda embl3 as the vector. screening of the library for the expression of m. pneumoniae protein antigens with adsorbed anti-m. pneumoniae serum revealed strong reactivity from a third of those clones which contained mycoplasma dna inserts. three of the most highly reactive clones were analyzed in detail and found to synthesize discrete mycoplasma proteins. two carried overlapping fragments of mycoplasma dna which en ... | 1986 | 3087877 |
| hydroxyl radical "footprinting": high-resolution information about dna-protein contacts and application to lambda repressor and cro protein. | a method has been developed for making "footprints" of proteins bound to dna. the hydroxyl radical, generated by reduction of hydrogen peroxide by iron(ii), is the reagent used to cut the dna. hydroxyl radical breaks the backbone of dna with almost no sequence dependence, so all backbone positions may be monitored for contact with protein. in addition to defining the dna sequence in contact with the protein, hydroxyl radical footprints embody structural information about the dna-protein complex. ... | 1986 | 3090544 |
| cloning of a cdna coding for human factor v, a blood coagulation factor homologous to factor viii and ceruloplasmin. | coagulation factor v is a high molecular weight plasma glycoprotein that participates as a cofactor in the conversion of prothrombin to thrombin by factor xa. a phage lambda gt11 hep g2 cell cdna expression library was screened by using an affinity-purified antibody to human factor v, and 11 positive clones were isolated and plaque-purified. the clone containing the largest cdna insert contained 2970 nucleotides and coded for 938 amino acids, a stop codon, and 155 nucleotides of 3' noncoding seq ... | 1986 | 3092220 |
| mr 26,000 antigen of schistosoma japonicum recognized by resistant wehi 129/j mice is a parasite glutathione s-transferase. | mice of the inbred strain 129/j bred at this institute (wehi 129/j) are relatively resistant to chronic infection with the parasitic helminth schistosoma japonicum. in contrast to more permissive mouse strains such as balb/c, the wehi 129/j mice are high responders to a mr 26,000 adult worm antigen designated sj26. cloned cdnas corresponding to sj26 have been identified in a s. japonicum phage lambda gt11 amp3 expression library, and their nucleotide sequences have been deduced. the predicted am ... | 1986 | 3095841 |
| overproduction from a cellulase gene with a high guanosine-plus-cytosine content in escherichia coli. | a recombinant exoglucanase was expressed in escherichia coli to a level that exceeded 20% of total cellular protein. to obtain this level of overproduction, the exoglucanase gene coding sequence was fused to a synthetic ribosome-binding site, an initiating atg, and placed under the control of the leftward promoter of bacteriophage lambda contained on the runaway replication plasmid vector pcp3 (e. remaut, h. tsao, and w. fiers, gene 22:103-113, 1983). with the exception of an inserted asparagine ... | 1986 | 3096205 |
| isolation and nucleotide sequence analysis of a cloned cdna encoding the beta-subunit of bovine follicle-stimulating hormone. | two different cdnas containing sequences coding for the beta-subunit of bovine follicle stimulating hormone (fsh-beta) have been isolated from a phage lambda gt11 bovine pituitary cdna library. the complete nucleotide sequence of both clones was determined, and the combined sequence represents most of fsh-beta mrna. the combined sequence contains 46 nucleotides of 5'-untranslated sequence followed by 387 nucleotides of coding sequence. the coding sequence predicts a 19-amino-acid amino-terminal ... | 1986 | 3096676 |
| in vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins. | an in vitro system was developed that provides a quick microscopic assay for nuclear transport. the assay uses an extract of xenopus eggs, normal or synthetic nuclei, and a fluorescently labeled nuclear protein, nucleoplasmin. this in vitro system accurately mimics in vivo nuclear transport, both in exclusivity and in the amount of accumulation observed (up to 17-fold). selective accumulation of fluorescent nucleoplasmin is observed microscopically within 30 min with rat liver nuclei, xenopus em ... | 1986 | 3097026 |
| cloning and expression of the isopenicillin n synthetase gene from penicillium chrysogenum. | the isopenicillin n synthetase (ips) gene from penicillium chrysogenum was isolated from a recombinant bacteriophage lambda library using the cephalosporium acremonium ips (cips) gene as a heterologous hybridization probe. the protein coding region of the p. chrysogenum ips (pips) gene was about 74% homologous to the cips gene, and the predicted amino acid sequences of the encoded proteins were about 73% homologous. escherichia coli cells with the pips gene contained ips activity whereas untrans ... | 1986 | 3104145 |
| cloning and characterization of the c1 repressor of pseudomonas aeruginosa bacteriophage d3: a functional analog of phage lambda ci protein. | we cloned the gene (c1) which encodes the repressor of vegetative function of pseudomonas aeruginosa bacteriophage d3. the cloned gene was shown to inhibit plating of d3 and the induction of d3 lysogens by uv irradiation. the efficiency of plating and prophage induction of the heteroimmune p. aeruginosa phage f116l were not affected by the presence of the cloned c1 gene of d3. when the d3 dna fragment containing c1 was subcloned into pbr322 and introduced into escherichia coli, it was shown to s ... | 1987 | 3106321 |
| regulation of the aroh operon of escherichia coli by the tryptophan repressor. | regulation of expression of aroh, the structural gene for the tryptophan-sensitive 3-deoxy-d-arabinoheptulosonic acid-7-phosphate synthetase, by the tryptophan repressor and its corepressor, l-tryptophan, was studied in vivo by using aroh-lacz fusions. protein and operon fusions were constructed on multicopy plasmids and subsequently crossed in single copy to the bacterial chromosome via the specialized transducing bacteriophage lambda rz5. analysis of the resulting lysogens demonstrated that ar ... | 1987 | 3106331 |
| unusual immunoglobulin dna sequences from the nonexpressed chromosome of mouse normal b lymphocytes: implications for allelic exclusion and the dna rearrangement process. | allelic exclusion of immunoglobulin gene products results in the expression of only one of two possible alleles in normal b lineage cells. attempts to define the role of heavy chain gene rearrangements in this process have revealed the nonexpressed allele to be rarely in its germline context, but rather incompletely rearranged (d/j rearrangements) or completely rearranged (v/d/j rearrangements) and in a context that cannot be expressed as a protein. nearly all dna sequences of heavy chain genes ... | 1987 | 3114375 |
| localization of the human thyroxine-binding globulin gene to the long arm of the x chromosome (xq21-22). | thyroxine-binding globulin (tbg) is the major thyroid-hormone transport protein in the plasma of most vertebrate species. a recombinant phage (lambda ctbg8) containing a cdna insert of human tbg recently has been described. with the cdna insert from lambda ctbg8 used as a radiolabeled probe, dna from a series of somatic-cell hybrids containing deletions of the x chromosome was analyzed by means of blot hybridization. the results indicated that the tbg gene is located in the midportion of the lon ... | 1987 | 3115094 |
| the rep region of pr plasmid regulates the expression of sos system. | by using an artificial hybrid between phage lambda and the pr plasmid, we have shown that the rep region of the pr plasmid encodes a function which regulates the expression of the muc genes (plasmid genes that are under the negative control of lexa and responsible for an increased rate of spontaneous mutagenesis and resistance to uv and chemicals). expression of the muc genes were monitored by a fusion between the muc promoter and the lacz structural gene. when e. coli cells containing such a fu ... | 1987 | 3118142 |
| immunogenic (tum-) variants of mouse tumor p815: cloning of the gene of tum- antigen p91a and identification of the tum- mutation. | mutagen treatment of mouse p815 tumor cells produces tum- variants that are rejected by syngeneic mice because these variants express new surface antigens. these "tum- antigens" are recognized by cytolytic t lymphocytes but induce no detectable antibody response. transfection of p815 cell line p1.htr with dna of tum- variant p91 yielded transfectants expressing tum- antigen p91a. they were detected by their ability to stimulate proliferation of cytolytic t lymphocytes [wölfel, t., van pel, a., d ... | 1988 | 3127830 |
| hormonal regulation of rat androgen-binding protein (abp) messenger ribonucleic acid and homology of human testosterone-estradiol-binding globulin and abp complementary deoxyribonucleic acids. | complementary dna clones coding for rat androgen-binding protein (rabp) were isolated from a rat testis cdna library constructed in the bacteriophage lambda gt11. the library was screened immunochemically, using two different antibodies against rabp. the identity of the isolated clones was confirmed by epitope selection and dna sequence analysis. the mrna encoding rabp could be detected in the testes of 20- and 46-day-old-rats, but not in the 10-day-old rats by hybridization with 32p-labeled rab ... | 1988 | 3135485 |
| primary structure of saccharomyces cerevisiae nadph-cytochrome p450 reductase deduced from nucleotide sequence of its cloned gene. | we isolated cdna (pgcyr, about 2.1 kb) and genomic dna (pggyr, about 4 kb) clones coding for nadph-cytochrome p450 reductase by immunoscreening of yeast saccharomyces cerevisiae cdna and genomic dna libraries in phage lambda gt11. the clones were sequenced and found to encode a protein of 691 amino acid residues with a calculated molecular weight of 76,737 daltons. the amino-terminal sequence (excluding the initial methionine residue) deduced therefrom was in agreement with the protein sequence ... | 1988 | 3139648 |
| human follicle-stimulating hormone beta-subunit gene encodes multiple messenger ribonucleic acids. | fsh is a pituitary gonadotropin that is encoded by separate alpha- and beta-subunit genes. we isolated a 12 kilobase (kb) dna fragment containing the entire human fsh beta gene from a lambda phage genomic dna library. the nucleotide sequence of the fsh beta gene predicts a 19 amino acid signal sequence and a 111 amino acid apoprotein that differs from the reported protein sequence at three residues and lacks the carboxyterminal eight amino acids, thereby bringing the human fsh beta sequence into ... | 1988 | 3139991 |
| effect of antiserum against aflatoxin b1-bovine serum albumin complex on aflatoxin b1-induced lysogenesis. | escherichia coli k12 bacteria lysogenic for the lambda phage were used to study the effect of antiserum against aflatoxin b1-induced lysogenesis. the antiserum was obtained from rabbits immunized with water in oil emulsion of aflatoxin b1-bovine serum albumin complex (afb1-bsa). a marked reduction in the degree of lysogenesis was observed when the antiserum was added to the reaction medium prior to microsomal enzyme activation of aflatoxin b1. there was no detectable effect when the antiserum wa ... | 1988 | 3140005 |
| mutant trp repressors with new dna-binding specificities. | oligonucleotide-directed mutagenesis of the codons for glutamine-68 (gln68), lysine-72 (lys72), isoleucine-79 (ile79), alanine-80 (ala80), and threonine-81 (thr81) of the escherichia coli trpr (tryptophan aporepressor) gene was used to make mutant repressors with each of 36 different amino acid changes. mutant repressors were tested for binding to each member of a set of 28 different operators closely related to the consensus trp operator. of the 36 mutant repressors, 11 bind a subset of the 28 ... | 1988 | 3140377 |
| human carbonyl reductase. nucleotide sequence analysis of a cdna and amino acid sequence of the encoded protein. | carbonyl reductase (ec 1.1.1.184) is one of several monomeric, nadph-dependent oxidoreductases having wide specificity for carbonyl compounds that are generally referred to as the aldoketoreductases. the grouping of the enzyme into the family has been proposed on the basis of functional similarities and in the absence of structural data. here, we describe the isolation and characterization of a cdna clone complementary to human carbonyl reductase mrna from a human placenta cdna library construct ... | 1988 | 3141401 |
| cloning of an outer membrane protein of neisseria meningitidis in escherichia coli. | a gene bank of chromosomal dna of neisseria meningitidis group b was constructed in phage lambda embl3, and screened by rabbit polyclonal antibodies to major outer membrane (om) proteins of the meningococcus. several clones expressing a 28 kda protein were found. the gene coding for the 28 kda protein was subcloned into plasmid puc18 in escherichia coli. the protein was expressed in e. coli and located in the om. rabbit antibodies were raised to the 28 kda protein purified from e. coli and used ... | 1987 | 3143889 |
| salmonella typhimurium lt2 metf operator mutations. | using an escherichia coli lac deletion strain lysogenized with lambda phage carrying a metf-lacz gene fusion (lambda flac), in which beta-galactosidase levels are dependent on metf gene expression, cis-acting mutations were isolated that affect regulation of the salmonella typhimurium metf gene. the mutations were located in a region previously defined as the metf operator by its similarity to the e. coli metf operator sequence. regulation of the metf gene was examined by measuring beta-galactos ... | 1988 | 3147373 |
| salmonella typhimurium mete operator-constitutive mutations. | we used a mete-lacz fusion phage (lambda elac) to select for mutants with operator-constitutive mutations in the salmonella typhimurium mete control region. all of the mutations identified were found to lie within a region containing tandemly-repeating 8-bp palindromes with the consensus sequence 5'-agacgtct-3', previously proposed to be the binding region for the metj-encoded repressor. lysogens carrying mutant lambda elac phage exhibit high beta-galactosidase levels that are only partially rep ... | 1988 | 3149604 |
| cloning and sequencing of cdna for the rat plasminogen activator inhibitor-1. | a cdna encoding rat plasminogen activator-inhibitor (pai-1) has been isolated from an htc rat hepatoma cell cdna library constructed in phage lambda gt10. the cdna contains 118 bp of 5'-untranslated sequence, 1206 bp encoding a 402-amino acid (aa) protein and 1747 bp of 3'-untranslated sequence. the protein-coding sequence and the derived amino acid sequence share 82% and 81% identity, respectively, with human pai-1 cdna and protein. the rat cdna encodes a preprotein with a 23-aa leader peptide ... | 1988 | 3149611 |
| molecular cloning of multiple xylanase genes from pseudomonas fluorescens subsp. cellulosa. | pseudomonas fluorescens subsp. cellulosa was shown to express extracellular xylanases. genes encoding these enzymes were isolated from a gene library of p. fluorescens subsp. cellulosa dna, constructed in bacteriophage lambda 47.1. one of the phages (pxc) that expressed xylanase also conferred the ability to hydrolyse carboxymethylcellulose. an 11.8 kb hindiii dna restriction fragment and a 6.2 kb ecori dna fragment were subcloned from two distinct xylanase-expressing phages, into puc18, to yiel ... | 1988 | 3151992 |
| mutation in ldl receptor: alu-alu recombination deletes exons encoding transmembrane and cytoplasmic domains. | the molecular size of the plasma ldl (low density lipoprotein) receptor synthesized by cultured fibroblasts from a patient with the internalization-defective form of familial hypercholesterolemia (fh 274) was smaller by 10,000 daltons than the size of the normal ldl receptor. the segment of the gene encoding the truncated portion of the fh 274 receptor was cloned into bacteriophage lambda. comparison of the nucleotide sequences of the normal and fh 274 genes revealed a 5-kilobase deletion, which ... | 1985 | 3155573 |
| reconstitution of recbc dnase activity from purified escherichia coli recb and recc proteins. | the escherichia coli recb protein, normally synthesized in low amounts, has been amplified by linkage of the recb gene to the phage lambda leftward promoter in an expression plasmid. from strains harboring this plasmid, recb protein has been purified to homogeneity by a simple procedure which includes affinity chromatography on a column of recc protein bound to agarose. the purified recb protein has dna-dependent atpase activity but no exonuclease activity. recc protein alone has neither atpase ... | 1985 | 3155726 |
| maximizing gene expression from plasmid vectors containing the lambda pl promoter: strategies for overproducing transcription termination factor rho. | we have constructed two plasmids in which transcription of the rho gene from escherichia coli k-12 is under the control of the lambda phage pl promoter. in p31-356, the normal rho promoter is deleted, but the remainder of the rho leader region, including the ribosome binding site, is present. in p39-as, the rho leader is completely absent, and the lambda cii ribosome binding site replaces that of rho. under noninducing conditions, expression of rho protein from these plasmids is repressed by the ... | 1985 | 3155859 |
| a phi 80 function inhibitory for growth of lambdoid phage in him mutants of escherichia coli deficient in integration host factor. ii. physiological analysis of the abortive infection. | derivatives of phage lambda with the rightmost 3% of the genome (the qsr region) from the related phage phi 80 fail to grow at low temperatures (e.g., 32 degrees) in escherichia coli hosts deficient in either protein component of ihf (integration host factor), the products of the hima and hip/himd genes. the abortive infection of lambda (qsr)80 in mutants defective for ihf was studied in detail. this infection is characterized by a lack of cell lysis and an inhibition of phage dna replication af ... | 1985 | 3155886 |
| [integration of the related prophages lambda, phi 80 and their hybrid lambda att80 into the secondary chromosomal att sites of wild-type escherichia coli]. | the family of lambdoid phages displays a varying specificity of integration into the host chromosome. the lambda phage dna failed to get inserted at the secondary attachment site(s) of the gal operon (frequency less than 2.6 x 10(-8)) in the presence of the primary (normal) one. by contrast, phi 80 and the lambda att80 hybrid integrated into wild-type escherichia coli at least, at two secondary att sites of the btub locus, the latter phage being also capable of integration in the vicinity of pur ... | 1985 | 3156067 |
| efficiency of induction of prophage lambda mutants as a function of reca alleles. | mutants of the ci gene of prophage lambda have been defined phenotypically in a reca+ host as noninducible (ind-), inducible (ind+), or induction sensitive (inds). we showed that a phage lambda ci+ carrying operator mutations v2 and v3 displays an inds phenotype, as does lambda ci inds-1. we characterized a fourth induction phenotype called induction resistant (indr). using these four prophage types, we tested the influence of bacterial reca mutations on prophage induction. indr prophages were f ... | 1985 | 3156121 |
| a human opal suppressor trna gene and pseudogene. | a human dna library, cloned in bacteriophage lambda, was screened with an opal suppressor trna probe. two genes were isolated, subcloned into pbr322, and sequenced. one is a normal opal suppressor trna gene 87 nucleotides in length without intervening sequences. it has a tca anticodon demonstrating that the mature trna reads the termination codon uga. the 5' internal control region for transcription has two extra nucleotides compared to the consensus sequence for eucaryotic trna genes, while the ... | 1985 | 3156131 |
| purification of a reca protein analogue from bacillus subtilis. | we have identified in bacillus subtilis an analogue of the escherichia coli reca protein. its activities suggest that it has a corresponding role in general genetic recombination and in regulation of sos (dna repair) functions. the b. subtilis protein (b. subtilis rec) has a mr of 42,000 and cross-reacts with antisera raised against e. coli reca protein. its level is significantly reduced in the recombination-deficient rece4 mutant. b. subtilis rec is induced 10- to 20-fold in rec+ strains follo ... | 1985 | 3156134 |
| interaction of the lambda site-specific recombination protein xis with attachment site dna. | nuclease protection experiments show that xis protein of bacteriophage lambda specifically binds attachment (att) site dna. the region of xis binding, present in both the phage att site and the right prophage att site, extends from position -102 to position -62 in the p arm. the sequence of this region, the positions of purines protected by xis against methylation, and the binding of xis to a resected att site indicate the presence of two binding sites. the postulated recognition elements, conta ... | 1985 | 3156374 |
| altered cro repressors from engineered mutagenesis of a synthetic cro gene. | a portion of the gene coding for the cro repressor protein of bacteriophage lambda has been chemically synthesized, incorporating base pair changes that generate restriction endonuclease sites without altering the amino acid coding sequence. these restriction endonuclease sites were used to remove small segments of the synthetic cro gene and the segments were replaced with duplexes carrying desired mutations. altered cro proteins produced by mutants constructed in this manner were then assayed f ... | 1985 | 3156377 |
| thermosensitivity of a dna recognition site: activity of a truncated nutl antiterminator of coliphage lambda. | antitermination is an important transcriptional control. in bacteriophage lambda, the presence of the nut antiterminators between the promoters and terminators results in relatively unhindered transcription when the lambda n gene product and necessary host factors are supplied. this antitermination system has been rendered thermosensitivity by modification of the nut site. a fragment of lambda dna [74 base pairs (bp) in length]that contained the 17-bp nutl core sequence, but lacked the 8-bp boxa ... | 1985 | 3156406 |
| involvement of the htpr gene product of escherichia coli in phage lambda development. | growth of phage lambda at high temperature requires a functional htpr host gene. the stages of the phage growth cycle shown to be dependent on htpr gene function include prophage excision and particle morphogenesis. two types of morphogenetic abnormalities have been detected. one is a defect in phage tail assembly that results from a deficiency in tail fibers even though gpj is produced. the severity of this defect is phage-strain specific. the second morphogenetic defect is less clearly defined ... | 1985 | 3156447 |
| protection of nonmodified phage lambda against ecok restriction mediated by reca protein. | a study was conducted to establish whether the ecok-specific restriction, which is alleviated in e. coli cells after uv induction of the sos response (day 1977), is also alleviated under the influence of an increased level of reca protein without induction of other sos functions. the host cells used were e. coli k-12, strain ab2497, and its derivatives; the nonmodified phage lambda was a mutant b2b5(vir). an increase of the reca protein level was induced using the plasmid px02, which is a recomb ... | 1985 | 3156795 |
| mutations in bacteriophage lambda repressor that prevent reca-mediated cleavage. | in this paper, we report on the isolation and sequence analysis of mutations that confer an induction-deficient phenotype to lambda repressor. a total of 16 different mutations, which occur at 13 different sites in the repressor gene, have been characterized. for most of the mutant lysogens, frequencies of spontaneous induction in a reca+ strain were reduced dramatically in comparison with those for a wild-type phage, and these mutant lysogens showed little or no prophage induction after uv irra ... | 1985 | 3156848 |
| expression of a gene for glucan-binding protein from streptococcus mutans in escherichia coli. | the structural gene for a glucan-binding protein (gbp) of streptococcus mutans has been inserted into a bacteriophage lambda vector and expressed in escherichia coli k12. lysates of e. coli infected with the recombinant phage contain an antigenic protein of the same size as s. mutans gbp. the gbp synthesized in e. coli can be affinity-purified on immobilized glucan and antiserum raised against it has been shown to precipitate fructosyltransferase activity from s. mutans. | 1985 | 3156960 |
| extent of sequence homology required for bacteriophage lambda site-specific recombination. | bacteriophage lambda integration and excision occur by reciprocal recombination within a 15-base homologous core region present in the recombining attachment (att) sites. strand exchange within the core occurs at precise nucleotide positions, which define an overlap region in which the products of recombination contain dna strands derived from different parents. in order to define the role of sequence homology during recombination we have constructed point mutations within the core and assayed t ... | 1985 | 3157003 |
| in phage lambda, cos is a recombinator in the red pathway. | among lambda particles carrying chromosomes that have failed to replicate during a lytic cycle cross there is a high frequency of red-mediated recombination near the right-hand end. earlier work has shown that this recombination is dependent on cos (cohesive end site), the packaging origin of lambda. in contrast to the prediction of the break-copy model proposed earlier, we find a high recombination rate near cos even when only one of the two participating parents has a functional cos at that lo ... | 1985 | 3157004 |
| the or control system of bacteriophage lambda. a physical-chemical model for gene regulation. | a quantitative model has been developed for processes in the bacteriophage lambda that control the switchover from lysogenic to lytic modes of growth. these processes include the interactions of ci repressor and cro proteins at the three dna sites of the right operator, or, the binding of rna polymerase at promoters pr and prm, the synthesis of ci repressor and cro proteins, and the degradative action of reca during induction of lysis. the model is comprised of two major physical-chemical compon ... | 1985 | 3157005 |
| weigle reactivation of phage lambda in a reca mutant of escherichia coli: dependence on the excess amounts of photoreactivating enzyme in the dark. | the plating efficiency of ultraviolet-light-irradiated phage lambda in the dark is increased when an escherichia coli reca host, which is transformed with a multicopy plasmid, pky1, carrying the phr gene of e. coli, is irradiated with 254 nm uv prior to infection (weigle (w) reactivation). such w reactivation in lexa3 and umuc strains, with or without pky1, is almost undetectable. addition of umu mutations to reca56/pky1 cells blocks this process, but addition of the lexa3 mutation instead gives ... | 1985 | 3157057 |
| lambda phage cro repressor interaction with its operator dna: 2'-deoxy-5-fluorouracil or3 analogues. | the experiments here show that chemically synthesized dna containing fluorine at selected sites can be used to test specific predictions of a model for cro repressor--operator interaction. this is done by observation of the perturbation to the fluorine-19 nmr spectra of analogues of or3 synthesized with 2'-deoxy-5-fluorouracil at specific positions in the dna helix. although the three-dimensional structure of the cro repressor from phage lambda has been determined by matthews and co-workers [and ... | 1985 | 3157402 |
| nh2-terminal arm of phage lambda repressor contributes energy and specificity to repressor binding and determines the effects of operator mutations. | several lines of evidence indicate that the phage lambda repressor recognizes its operator by using, in part, an alpha helix (the "recognition helix"), which it inserts into the major groove of dna. in addition to its recognition helix, lambda repressor has an "arm," consisting of the first six amino acids, that wraps around the dna helix. we constructed plasmids that, in escherichia coli, direct the expression of derivatives of lambda repressor that lack the nh2-terminal one, three, six, or sev ... | 1985 | 3157988 |
| characterization of the spo0a locus and its deduced product. | the highly pleiotropic stage 0 sporulation locus of bacillus subtilis, spo0a, has been cloned in bacteriophage lambda, subcloned in plasmids, and sequenced. the locus was found to code for a protein of 29,691 da. analysis of the in vivo transcripts from this region by nuclease s1 protection experiments located the start and stop of transcription of the locus. the transcription start site was preceded by a promoter resembling sigma 37-dependent promoters. two mutations originally assigned to a se ... | 1985 | 3157992 |
| propagation of some human dna sequences in bacteriophage lambda vectors requires mutant escherichia coli hosts. | the growth of clones of human genomic dna fragments in a bacteriophage lambda vector has been examined in a number of different escherichia coli hosts. a large proportion (8.9%) of the phages carrying different fragments of the human genome fail to grow on standard rec+ hosts but will grow on hosts carrying mutations in the recb, recc, and sbcb genes. heteroduplex analysis in the electron microscope of dna from four of these phages revealed substantial secondary structure, including snap-back re ... | 1985 | 3157994 |
| rec-mediated recombinational activity of two adjacent chi elements in bacteriophage lambda. | 1985 | 3158572 | |
| involvement of dna polymerase iii in uv-induced mutagenesis of bacteriophage lambda. | it has been proposed that the mutation fixation processes stimulated by sos induction result from an induced infidelity of dna replication (radman 1974). the aim of this study was to determine if mutator mutations in the e. coli dna polymerase iii might affect uv-induced mutagenesis. using a phage lambda mutation assay which can discriminate between targeted and untargeted mutations, we show that the polc74 mutator mutation (sevastopoulos and glaser 1977) primarily affects untargeted mutagenesis ... | 1985 | 3158798 |
| [regions of human genome-containing analogs of oncogenes and retroviral genes. ii. a new class of retrovirus-like elements]. | earlier we have found that recombinant phage lambda gp5 contains the sequences homologous to v-mos oncogene and retroviruses. after the nucleotide sequence determination we have found the region with homology to u5 part of retroviral ltr. adjacent to this region are sequences complementary to 3'-end of trnamet. numerous transcripts reacting with subcloned u5 probe from gp5 are present in polyadenylated rna fraction from human cells. the humane genome contains several copies of these region, with ... | 1985 | 3158802 |
| bacteriophage lambda vector for transducing a cdna clone library into mammalian cells. | we have developed a bacteriophage lambda vector (lambda nmt) that permits efficient transduction of mammalian cells with a cdna clone library constructed with the pcd expression vector (h. okayama and p. berg, mol. cell. biol. 3:280-289, 1983). the phage vector contains a bacterial gene (neo) fused to the simian virus 40 early-region promoter and rna processing signals, providing a dominant-acting selectable marker for mammalian transformation. the phage dna can accommodate pcd-cdna recombinants ... | 1985 | 3158804 |
| the extent of dna sequence required for a functional bacterial attachment site of phage lambda. | we have investigated the extent of dna sequence required to form a bacterial attachment site (attb) that functions in bacteriophage lambda integration. a dna fragment carrying attb of escherichia coli was trimmed, recloned and tested for recombination proficiency. we found that the common core sequence plus the adjoining 4-bp sequences of both the b and b' arms are required for full activity, while plasmids with an even shorter attb sequence retain some capacity to function as attb in vivo. we a ... | 1985 | 3158879 |
| structure of the galactokinase gene of escherichia coli, the last (?) gene of the gal operon. | we present the nucleotide sequence of the galactokinase gene (galk) of escherichia coli including its 5' and 3' flanking regions. this dna sequence derives from the lambda gal8 transducing phage and is identical to the sequence present in the galk gene fusion vectors, pko and pkg, commonly used to study transcriptional regulatory elements. we define the precise 3' junction between the bacterial and phage sequences in lambda gal8 and demonstrate that this junction probably results from a homologo ... | 1985 | 3158881 |
| efficient modification of e. coli rna polymerase in vitro by the n gene transcription antitermination protein of bacteriophage lambda. | the n gene protein of bacteriophage lambda prevents termination of transcription by e. coli rna polymerase. we describe here the conditions of a cell-free reaction system in which pure n stimulates net transcription up to tenfold and therefore nearly stoichiometrically modifies transcribing rna polymerase molecules. the reaction contains micrococcal nuclease-treated s100 extract derived from e. coli and a plasmid template dna containing the lambda early promoter pl, the n utilization site nutl, ... | 1985 | 3158883 |
| overproduction and purification of protein p6 of bacillus subtilis phage phi 29: role in the initiation of dna replication. | a phi 29 dna fragment containing gene 6, required for dna replication, has been cloned in plasmid pplc28 under the control of the pl promoter of phage lambda. a polypeptide with an electrophoretic mobility close to that of p6 was labelled with 35s-methionine after heat induction. this protein, representing about 4% of the total e. coli protein after 1 h of induction, was obtained in a highly purified form. the protein was characterized as p6 by amino acid analysis and nh2-and cooh-terminal seque ... | 1985 | 3158884 |
| genetic rearrangement of dna induces knots with a unique topology: implications for the mechanism of synapsis and crossing-over. | we have determined the topological sign of the knots produced by a cycle of phage lambda integrative recombination. to insure that these knots reflect intrinsic features of the reaction mechanism, the substrate was constructed so that random interwrapping of segments of dna played a minimal role in the topological outcome. the knotted dna was coated with the bacteriophage t4 uvsx gene product and examined in the electron microscope to determine the nature of each crossing point or node. all of t ... | 1985 | 3159013 |
| a cii-dependent promoter is located within the q gene of bacteriophage lambda. | we have found a cii-dependent promoter, paq, within the q gene of bacteriophage lambda. transcription experiments and abortive initiation assays performed in vitro showed that the promoter strength and the cii affinity of paq were comparable to the other cii-dependent lambda promoters, pe and pi. the location and leftward direction of paq suggests a possible role in the delay of lambda late-gene expression by cii protein, a phenomenon that has been called cii-dependent inhibition. we have constr ... | 1985 | 3159014 |
| dual role for escherichia coli reca protein in sos mutagenesis. | induction of the escherichia coli sos system increases the ability of the cells to perform dna repair and mutagenesis. previous work has shown that this increased mutagenesis is the result of derepression of specific genes through a complex regulatory mechanism controlled by lexa and reca proteins. one role of reca protein in this process is to facilitate proteolytic cleavage of lexa protein (the repressor) in response to an inducing signal that reversibly activates reca protein to perform this ... | 1985 | 3159017 |
| functional and physical characterization of transcription initiation complexes in the bacteriophage lambda or region. | we have used transcriptional activity assays and dnase i footprinting techniques to examine in vitro the binding of escherichia coli rna polymerase and lambda repressor protein to the bacteriophage lambda rightward promoter-operator region. for the lambda pr promoter, the activity and physical binding results determined at several repressor concentrations correlated very well. good agreement was also found for repression of prm, which occurred at higher repressor concentrations; however, our res ... | 1985 | 3159734 |
| an escherichia coli mutant unable to support site-specific recombination of bacteriophage lambda. | we report the isolation of mutations in, and the characterization of, an escherichia coli gene, hip, that is required for site-specific recombination of phage lambda. hip mutants are recessive and are located near minute 20 on the linkage map. the gene product is not vital to bacterial growth, since deletion mutants are viable. the absence of hip product reduces lambda integration to barely detectable levels and also reduces prophage excision, but less drastically. certain mutations in the lambd ... | 1985 | 3159904 |
| sos induction by thermosensitive replication mutants of minif plasmid. | minif, a 9.3 kb fragment of the dispensable f plasmid, carries genes necessary for its replication and partition as well as for the expression of an sos signal. the arrest of replication of a thermo-sensitive minifts at 42 degrees c induced sos functions such as prophage lambda, sfia expression, w-reactivation of uv-irradiated phage lambda. two minif ts9 and ts17 mutations were located within the kpni fragment (43.6-46.9) in the minimal oris replicon. blocking minif replication by incbc+ incompa ... | 1985 | 3159950 |
| an in vitro test for photoinduced toxicity of benzothiadiazine diuretics using bacteriophage lambda. | 1985 | 3160053 | |
| the stereostructure of knots and catenanes produced by phage lambda integrative recombination: implications for mechanism and dna structure. | we studied the mechanism of recombination by determining the structure of the products of the phage lambda int system. electron microscopy of reca-coated products revealed only knots and catenanes containing a regular right-handed spiral structure. the structure and distribution of products establish that the recombination sites pair by essentially random collision, rather than by tracking. however, the distribution also indicates that the binding of the enzyme must introduce nonrandom component ... | 1985 | 3160481 |
| 1h nmr study of the interaction of bacteriophage lambda cro protein with the or3 operator. ii. assignment of the non-exchangeable proton resonances of the or3 operator. | the 17 base pair operator or3 oligonucleotide, which is the preferential binding site for the cro repressor of phage lambda, was studied by two-dimensional nmr spectroscopy. a sequential assignment procedure based on two-dimensional nuclear overhauser effect (noesy) and scalar coupling correlated (cosy) nmr spectroscopy, together with the knowledge of the oligodesoxynucleotide sequence, made it possible to assign the non-exchangeable base protons and the h1' and the h2'-h2" sugar protons of the ... | 1985 | 3160577 |
| inhibitory effect of high-level transcription of the bacteriophage lambda nutl region on transcription of rrna in escherichia coli. | transcription of the bacteriophage lambda nutl region from the pl promoter on a multicopy plasmid in escherichia coli causes a reduction in growth rate and in transcription of rrna relative both to total transcription and to transcription of trnas that are not encoded in rrna operons. these observations support the hypothesis, previously based on nut site dna sequence homology, that the phage lambda and rrna antitermination systems are related. | 1985 | 3160688 |
| direct selection of mutations reducing transcription or translation of the reca gene of escherichia coli with a reca-lacz protein fusion. | when a reca-lacz protein fusion was cloned into phage lambda, the resulting transducing phage grew normally on wild-type escherichia coli, but its growth was severely inhibited in lexa(def) mutant strains that express reca constitutively at high levels. mutants of the transducing phage that grew on the lexa(def) strains were isolated and were found to affect production of the reca-beta-galactosidase hybrid protein. most mutants, including a number of nonsense mutants, were phenotypically lacz-. ... | 1985 | 3160689 |
| interaction of rho factor with bacteriophage lambda cro gene transcripts. | rho protein is responsible for termination of transcription of the cro gene of bacteriophage lambda. since rho is known to interact with the rna whose synthesis is being terminated, we measured the specificity and strength of binding of rho to isolated cro transcripts, using a nitrocellulose filter retention assay. the association constant (k alpha) for the binding of rho to a 372-nucleotide cro transcript was determined to be 7 +/- 2 x 10(8) m-1 at 37 degrees c and about 20-fold less at 4 degre ... | 1985 | 3160698 |
| homology requirements for recombination in escherichia coli. | the dna sequence homology required for recombination in escherichia coli has been determined by measuring the recombination frequency between insulin dna in a miniplasmid pi vx and a homologous sequence in a bacteriophage lambda vector. a minimum of approximately equal to 20 base pairs in a completely homologous segment is required for significant recombination. there is an exponential increase in the frequency of recombination when the length of homologous dna is increased from 20 base pairs to ... | 1985 | 3161076 |
| defective antitermination of rrna transcription and derepression of rrna and trna synthesis in the nusb5 mutant of escherichia coli. | the nusb5 mutant of escherichia coli was originally selected for reduced ability to support the antitermination of transcription that is mediated by the gene n product of bacteriophage lambda. by analyzing pulse-labeled rna with an rna.dna filter hybridization technique, we have shown that, in the nusb5 mutant, the ratio of promoter-proximal rrna transcripts to promoter-distal transcripts is increased at least by a factor of 1.6; that is, in the absence of the functional nusb gene product, prema ... | 1985 | 3161080 |
| [lethal and mutagenic action of incorporated 5-3h-cytosine on extracellular phage lambda]. | a study was made of the lethal and mutagenic effects on extracellular phage gamma of 5-3h-cytosine incorporated into dna. the efficiencies of inactivation by incorporated 3h were equal for 5-3h-cytosine and [3h-methyl]-thymidine, but the yield of c-mutations for the former was 14 times higher. the lethal and mutagenic effects of incorporated 5-3h-cytosine did not depend on ung mutation of host cells which caused a deficiency in uracil-dna-glycosylase. the mutagenic effect was not enhanced when s ... | 1985 | 3161117 |
| reinitiation during lambda dna replication resulting from either cis-pt treatment or infection of a p2 lysogenic strain. | nested areas of replication are observed in phage lambda replicative intermediates and arise from reinitiation from the lambda origin. reinitiation occurs when the first round of lambda replication takes place in the presence of the drug cis-pt or when lambda infects a host which has been preincubated with the drug. in the latter case it is shown that the infection proceeds during the expression of sos functions induced in the host as a result of the drug treatment. when lambda infects a host ly ... | 1985 | 3161238 |
| isolation of lambda phage dna by hydroxylapatite chromatography. | a simple and rapid (1 day) method for preparation of lambda phage dna was proposed. the method included two main steps: (a) growth and lysis of bacteria containing lambda phage and (b) purification of lambda phage dna by hydroxylapatite chromatography. the phage dna prepared by this method was intact and free of rna, proteins, and bacterial dna. | 1985 | 3161411 |
| large macromolecules can be introduced into cultured mammalian cells using erythrocyte membrane vesicles. | plasmid 6.4 kbp dna, 14 kbp dna, lambda phage particles, all of which contained herpes simplex virus type 1 (hsv-1) thymidine kinase (tk) gene, or igm molecules, were mixed with erythrocyte membranes and treated with neutral detergent. the transparent mixture was diluted with phosphate-buffered saline (pbs), followed by centrifugation to collect membrane vesicles containing the large macromolecules. 10-15% of 6.4 kbp, 3% of 14 kbp, 4-7% of the lambda phage particles and 14.5% of igm were trapped ... | 1985 | 3161750 |
| cloning of the newt pleurodeles waltlii chromosomal dna. | pleurodeles waltlii genomic dna has been cloned using several phage lambda vectors. we have isolated approx. 600 000 clones, which correspond to about 20% of the total dna sequences of this organism. this constitutes the first large gene library of a urodele. the low yield of cloning was attributable to the abundance of highly repetitive sequences, since recombinations in the bacterial host could lead to the loss of clones. indeed, the existence of highly repetitive sequences was directly demons ... | 1985 | 3161783 |
| isolation and characterization of the glucose-6-phosphate dehydrogenase gene of drosophila melanogaster. | to investigate the molecular basis of dosage compensation in drosophila, a recombinant lambda phage containing the drosophila melanogaster glucose-6-phosphatase dehydrogenase (g6pd) gene was isolated by differential screening of a drosophila genomic lambda library with poly(a)+rna obtained from polyribosomes enriched for or depleted of g6pd mrna sequences. of 44 000 plaques screened, a single phage, lambda dmg21, showed hybridization with the enriched poly(a)+rna but not the depleted one. confir ... | 1985 | 3161784 |
| control of phenylalanyl-trna synthetase genetic expression. site-directed mutagenesis of the phes, t operon regulatory region in vitro. | previous studies of phenylalanyl-trna synthetase expression in escherichia coli strongly suggested that the phes, t operon was regulated by a phenylalanine-mediated attenuation mechanism. to investigate the functions of the different segments composing the phes, t attenuator site, a series of insertion, deletion and point mutations in the phes, t leader region have been constructed in vitro on a recombinant m13 phage. the effects of these alterations on the regulation of the operon were measured ... | 1985 | 3162032 |
| gene fusions to the ptsm/pel locus of escherichia coli. | we have constructed gene fusions between ptsm/pel and lacz. these fusions affect both phenotypes assigned to the ptsm/pel locus (at 40 min), namely, no growth on mannose or glucosamine and inhibition of the penetration of bacteriophage lambda dna, as well as that of other lambdoid phages such as hy-2. since the lacz gene fusions are insertion mutations that abolish target gene function by disrupting the linear contiguity of the gene, it would appear that ptsm and pel are either the same gene or ... | 1985 | 3162078 |
| induction of 3-methyladenine dna glycosylase ii is reca+-independent. | the reca1 mutation was transduced into the tag-2 mutant of e. coli, thus making a strain deficient in the induction of sos repair as well as in the constitutive repair of 3-alkylated adenines in dna. the double mutant reca tag is more sensitive to methyl methanesulfonate exposure than either single mutant, indicating that reca and tag mutations block different pathways in repair of alkylation damage. the double mutant is more deficient in host cell reactivation of alkylated phages than the tag s ... | 1985 | 3162098 |
| sequence and deletion analysis of the recombination enhancement gene (ref) of bacteriophage p1: evidence for promoter-operator and attenuator-antiterminator control. | the ref gene of bacteriophage p1 stimulates recombination between two defective lacz genes in the escherichia coli chromosome (lac x lac recombination) and certain other reca-dependent recombination processes. we determined the dna sequence of the 5' portion of the ref gene and tested various regions for functionality by inserting dna fragments lacking increasing amounts of 5' sequence into plasmid and lambda phage vectors and measuring the ability of the constructs to stimulate lac x lac recomb ... | 1988 | 3170487 |
| structure and expression of an actin gene of physarum polycephalum. | physarum polycephalum (strain m3cviii) contains four unlinked actin gene loci, each with two alleles (arda1, arda2, ardb1, ardb2, ardc1, ardc2, ardd1 and ardd2). the 4800 base hindiii fragment of the ardc2 allele was previously isolated as a recombinant phage lambda. we now report the structure of the actin gene sequences (c-actin gene). the gene, which contains four intervening sequences, codes for the principal actin isotype of plasmodia and it is expressed in both the haploid myxamoebal and d ... | 1988 | 3172209 |
| transcriptional regulation of early functions of bacteriophage phi 80. | to study the expression of early functions of phi 80 phage, various segments from the early region were transcribed with rna polymerase. two major transcripts (from promoters pl and pr) whose synthesis was inhibited by the ci protein were identified. synthesis of the third major transcript (from promoter prm) was induced by the ci protein. these studies define two operator-promoter regions, olpl and orprprm. this mode of transcription from the early region is similar to that of phage lambda. how ... | 1988 | 3172226 |
| characterization of the cdna encoding a protein binding to the major histocompatibility complex class ii y box. | the expression of hla class ii genes is regulated by a series of cis-acting elements and trans-acting factors. several cis-acting elements have been identified and have been termed the z box, x box, y box, octamer, and "tata" box. the y box contains an inverted ccaat box. by probing a phage lambda gt11 library with double-stranded oligonucleotides, we have directly isolated a cdna encoding a y box-binding protein designated yb-1. yb-1 binding has an absolute requirement for the ccaat box and rel ... | 1988 | 3174636 |
| characterization of cdnas, mrnas, and proteins related to human liver microsomal cytochrome p-450 (s)-mephenytoin 4'-hydroxylase. | a cytochrome p-450 (p-450) multigene family codes for several related human liver enzymes, including the p-450 responsible for (s)-mephenytoin 4'-hydroxylation. this enzyme activity has previously been shown to be associated with a genetic polymorphism. genomic (southern) blot analysis using non-overlapping 5' and 3' portions of a cdna clone suggests that approximately seven related sequences are present in this gene family. in this study four cdna clones, all nearly full-length, were isolated f ... | 1988 | 3196692 |
| molecular cloning of a salmonella typhi lt-like enterotoxin gene. | diarrhoea is a common event during typhoid fever; nevertheless, the possible participation of a diarrhoea-inducing enterotoxin has not been described (roy et al., 1985). recombinant bacteriophage lambda fdc1 was isolated from a genomic library of salmonella typhi, the causal agent of typhoid fever, by screening with a probe for the b subunit gene of the heat-labile, cholera-like, escherichia coli enterotoxin (lt). lambda fdc1 codes for an enterotoxin that causes secretion in rat ileal loops, tha ... | 1988 | 3210968 |
| mulcos: a vector for amplification and simultaneous expression of two foreign genes in mammalian cells. | a method was developed for amplification and expression of foreign genes in mammalian cells. this procedure exploits the fact that an sfii cleavage site, ggccgcct/cggcc (the recognition sequences are underlined), is present at the sv40 replication origin and the cleaved ends, cct-3' and agg-3', are not rotationally equivalent. thus dna fragments flanked by the sfii sites can be ligated in head-to-tail tandem arrays and cloned in cosmids; the resulting construct is called a mulcos. the cosmid vec ... | 1988 | 3215524 |
| cloning and comparative sequence analysis of the gene coding for isopenicillin n synthase in streptomyces. | the genes coding for isopenicillin n synthase (ipns) in streptomyces jumonjinensis and s. lipmanii were isolated from recombinant phage lambda libraries using the s. clavuligerus ipns gene as a heterologous probe. the s. jumonjinensis ipns gene has an open reading frame coding for 329 amino acids, identical in size to that of the previously cloned s. clavuligerus ipns gene. a partial nucleotide sequence was also determined for the s. lipmanii ipns gene. comparison of the predicted amino acid seq ... | 1988 | 3216857 |
| the rat androgen receptor: primary structure, autoregulation of its messenger ribonucleic acid, and immunocytochemical localization of the receptor protein. | a composite androgen receptor dna sequence 4,181 base pairs in length was determined from three cdna clones isolated from a rat epididymal bacteriophage lambda gt11 library. an open reading frame of 902 amino acids encodes a protein of 98,227 mol wt. structural domains characteristic of the steroid receptor family include an amino-terminal region with five repeated amino acid motifs, a central dna-binding domain homologous with other steroid receptors, and a carboxyl-terminal steroid-binding reg ... | 1988 | 3216867 |
| 'conservative' and 'variable' clusters of alu-family dna repeats in human chromosomes. | the distribution of alu-family dna repeats (afrs) in chromosomes of phytohaemagglutinin-stimulated peripheral blood lymphocytes of four normal donors and non-stimulated bone marrow cells of four patients with acute leukemia (all and anll) was studied by in situ hybridization using dna of recombinant phage lambda containing multiple inserts of afr as a probe. over some chromosome bands (14cen, 16p13, 16cen) from normal donors and from leukemic patients clusters of silver grains were detected. ove ... | 1988 | 3221844 |
| species-specific biotinylated dna probe for the detection of mycoplasma gallisepticum. | a 5.5 kilobase dna fragment from an eco ri digest of the mycoplasma gallisepticum genome was specific for the detection of m. gallisepticum. this 5.5 kb fragment was initially cloned into bacteriophage lambda gt11 followed by subcloning into the plasmid vector pgem-3z. the incorporation of a biotin label was accomplished by utilizing biotin-11-dutp in a nick translation reaction. this probe, designated pmg6, reacts specifically with m. gallisepticum when tested against various mycoplasma dnas in ... | 1988 | 3221885 |
| [differences in the localization of alu-repeats in various chromosomes of peripheral blood cells from normal donors and bone marrow cells from patients with acute leukemia]. | the dispersion of the alu-family dna repeats in phytohemagglutinin-stimulated lymphocytes from peripheral blood of normal donors as well as in nonstimulated bone marrow cells of four patients suffering from acute leukemia was studied by hybridization on metaphase chromosomes in situ. dna of bacteriophage lambda car42 clone containing the insertion of at least 8 copies of alu-family dna-repeats and labelled with tritium was used as a probe in hybridization. all patients with acute leukemia had th ... | 1988 | 3237236 |
| the transcriptionally active human ribosomal protein s17 gene. | a human ribosomal protein s17 cdna [chen et al., proc. natl. acad. sci. usa 83 (1986) 6907-6911] was used to isolate four s17 dna clones from human genomic libraries constructed in bacteriophage lambda and cosmid vectors. based on its transcriptional activity in a transient expression assay and on sequence similarity with s17 cdna, cosmid clone hgs17-6 was identified as carrying the functional rps17 gene. rps17 is composed of five exons and four introns that span 4 kb of dna. two lambda clones o ... | 1988 | 3240863 |