Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| purification of the phou protein, a negative regulator of the pho regulon of escherichia coli k-12. | thermally induced transcription of the phou gene under control of the major leftward promoter, pl, of phage lambda resulted in production of the phou protein to compose approximately 5% of the total cell protein. the phou protein was present in the cytoplasm in the form of an aggregate. the amino acid composition and n-terminal amino acid sequence of the purified protein confirmed the reading frame established earlier for the phou gene. | 1986 | 3536855 |
| bacterial alkaline phosphatase clonal variation in some escherichia coli k-12 phor mutant strains. | several phor alleles (phor19, phor20, phor68, phor69, phor70, and phor78) led to either a bacterial alkaline phosphatase (bap)-constitutive phenotype or a variable behavior, depending upon the strain tested. whereas escherichia coli k10, mc1000, and xph4 phor mutants were constitutive, ab1157, bd792, mc4100, and w3110 phor mutants displayed the metastable character. for the latter strains, constitutive mutants regularly segregated bap-negative clones which yielded constitutive variants again at ... | 1986 | 3536875 |
| [a new vector for cloning of genes and replicons in yeasts]. | a novel escherichia coli-saccharomyces cerevisiae shuttle vector lambda man78 has been constructed. the vector contains phage lambda 47.1 dna, sacch. cerevisiae chromosomal segment with trp1 gene and the yeast ars1 replicator. this vector may be propagated as a phage and, similar to parental lambda 47.1, allows direct selection of large dna inserts (15-24 kbp) in e. coli. lambda man78 can efficiently transform licl-treated yeast cells (3-5 x 10(3) transformants per 1 microgram dna). replication ... | 1986 | 3542710 |
| characterization of a rat liver cytochrome p-450ut-h cdna clone and comparison of mrna levels with catalytic activity. | a rat liver cdna library was prepared using the expression vector bacteriophage lambda gt11 and plaques were screened using polyclonal antibodies raised to purified rat liver cytochrome p-450ut-h, the major enzyme involved in debrisoquine 4-hydroxylation, bufuralol 1'-hydroxylation, and sparteine delta 5-oxidation. a clone was selected which contained a 1.3-kb insert. the escherichia coli beta-galactosidase fusion protein had a molecular weight greater than that of native beta-galactosidase (and ... | 1987 | 3543648 |
| cloning and expression in escherichia coli of a synthetic dna for hirudin, the blood coagulation inhibitor in the leech. | a 235-bp dna coding for the leech blood coagulation inhibitor, hirudin, was chemically synthesized. the synthesis involved preparation of seven long oligodeoxyribonucleotide pairs which were assembled and cloned using a rapid and simple procedure. more than half of the transformed escherichia coli cells expressed a biosynthetic polypeptide having biological properties which were very similar to authentic hirudin from the leech hirudo medicinalis. to achieve efficient expression, we fused the hir ... | 1986 | 3545721 |
| cloning and expression in escherichia coli of the ibc protein genes of group b streptococci: binding of human immunoglobulin a to the beta antigen. | total-cell dna isolated from a highly virulent serotype ic strain of a group b streptococcus was used to construct a gene bank with bacteriophage lambda l47.1 in escherichia coli k-12. recombinant phage plaques in the bank were immunoblotted by using anti-alpha- and anti-beta-specific antibodies directed towards the ibc proteins purified from the streptococcal cell surface, and hybrid phages expressing the alpha protein (lambda alpha +) and the beta protein (lambda beta +) were identified. dna i ... | 1987 | 3552990 |
| nucleotide sequence and transcription of a human trna gene cluster with four genes. | a bacteriophage lambda clone containing a 20-kb human dna segment was isolated and found to harbor a cluster of four trna genes. an 8.2-kb hindiii subfragment encompassing the genes was cloned into pbr322 for restriction mapping and dna sequence analysis. the genes were found to be arranged as two tandem pairs, separated by 3 kb. a proline trnaagg gene is separated from a leucine trnaaag gene by a 724-bp intergenic region in the first pair, and a second proline trnaagg gene is 316 bp from a thre ... | 1986 | 3557125 |
| the cdna-deduced primary structure of human sex hormone-binding globulin and location of its steroid-binding domain. | we have sequenced a cdna for sex hormone-binding globulin (shbg) isolated from a phage lambda gt11 human liver cdna library. the library was screened with a radiolabeled rat androgen-binding protein (abp) cdna, and the abundance of shbg cdnas was 1 in 750,000 plaques examined. the largest human shbg cdna (1194 base-pairs) contained a reading frame for 381 amino acids. this comprised 8 amino acids of a signal peptide followed by 373 residues starting with the known nh2-terminal sequence of human ... | 1987 | 3569533 |
| 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 |
| dna sequence analysis of the dutch beta zero-thalassemia deletion. | the dutch beta zero-thalassemia has relatively few clinical symptoms in the homozygote. heterozygotes have levels of fetal hemoglobin (4-11%) comparable to delta beta-thalassemia, and much higher than in typical beta zero-thalassemia. to analyze this deletion, we have cloned the abnormal 10 kb bgl ii fragment that contains the delta-globin gene into phage lambda vector embl4. a hind iii-xba i fragment that includes the endpoints of the deletion was subcloned into plasmid puc 19 and partially seq ... | 1987 | 3593292 |
| interspersion of histone and 5s rna genes in artemia. | four recombinant lambda phage containing histone genes were selected from a library of artemia genomic dna fragments. the histone gene organization of artemia resembles that of other invertebrates in that all five genes are clustered and repeated in tandem with approximate repeat lengths of 8.5 kb and 9.3 kb. each recombinant lambda phage isolate hybridizes with five histone mrnas and unexpectedly also with 5s ribosomal rna. hybridization kinetics have shown the number of histone genes to be abo ... | 1987 | 3596239 |
| strand scissions of dna by patulin in the presence of reducing agents and cupric ions. | in the presence of nadph and cucl2, patulin induced the cleavage of cole1 dna and lambda phage dna in vitro. the dna-cleaving activity of patulin was concentration dependent. at the lowest concentration of patulin, cole1 supercoiled dna was relaxed and the highest concentration induced linearization of the dna. this activity was inhibited by superoxide dismutase, catalase and radical scavengers, showing involvement of free radicals in the dna-cleavage. lambda phage dna was also degraded by patul ... | 1987 | 3610824 |
| selection of dna binding sites by regulatory proteins. statistical-mechanical theory and application to operators and promoters. | we present a statistical-mechanical selection theory for the sequence analysis of a set of specific dna regulatory sites that makes it possible to predict the relationship between individual base-pair choices in the site and specific activity (affinity). the theory is based on the assumption that specific dna sequences have been selected to conform to some requirement for protein binding (or activity), and that all sequences that can fulfil this requirement are equally likely to occur. in most c ... | 1987 | 3612791 |
| bacteriophage 82 gene q and q protein. sequence, overproduction, and activity as a transcription antiterminator in vitro. | phage 82 gene q encodes a phage-specific positive regulator of late gene expression, thought, by analogy to the corresponding gene of phage lambda, to be a transcription antiterminator. we have cloned and sequenced the phage 82 gene q and have overproduced and purified the 82 q protein. we also have identified and sequenced dna containing the phage 82 late gene promoter and terminator. we show that purified 82 q protein is active and specific for dna containing the 82 late gene promoter in a wel ... | 1987 | 3624233 |
| structure of a gene encoding the 1.7 s storage protein, napin, from brassica napus. | a rapeseed chromosomal region containing a gene (napa), which encodes the 1.7 s seed storage protein (napin), was isolated in several overlapping recombinant clones from a phage lambda genomic library. following restriction enzyme mapping of the genomic region, a subclone containing the napa coding region as well as some 1.1 and 1.4 kilobases of dna from the 5' and 3' regions, respectively, was mapped and sequenced. the gene turned out to lack introns. southern blotting analyses utilizing a napi ... | 1987 | 3624251 |
| cloning and sequencing of the medium-chain s-acyl fatty acid synthetase thioester hydrolase cdna from rat mammary gland. | cdna clones coding for the medium-chain s-acyl fatty acid synthetase thioester hydrolase (thioesterase ii) from rat mammary gland were identified in a bacteriophage lambda gt11 library and their nucleotide sequences were determined. the predicted coding region spans 263 amino acid residues and includes a sequence identical with that of a peptide derived from the enzyme active site. the rat thioesterase ii cdna sequence exhibits homology with that of a thioesterase found in duck uropygial glands. | 1987 | 3632637 |
| two human tyrosine trna genes contain introns. | a human lambda-phage recombinant which contains at least four trna genes, has been isolated. two dna fragments were subcloned to give the recombinant plasmids pm6 and pm6128. nucleotide sequence analysis showed that each plasmid contained a different tyrosine acceptor trna (trnatyr) gene. both trnatyr genes are interrupted by 21-bp introns. these recombinant plasmids have been shown to direct the in vitro synthesis of trna-sized products in a hela cell transcription system. | 1986 | 3636257 |
| effects of 3'-(3-cyano-4-morpholinyl)-3'-deaminoadriamycin and structural analogues on dna in ht-29 human colon carcinoma cells. | the potent adriamycin (adr) analogue, 3'-(3-cyano-4-morpholinyl)-3'-deaminoadriamycin (cma), produces dna-dna cross-links in human and murine tumor cells. the cellular pharmacology of cma, its derivative, 5-imino-3'-(3-cyano-4-morpholinyl)-3'-deaminoadriamycin (icma), 3'-(4-morpholinyl)-3'-deaminoadriamycin (ma), and adr was evaluated in ht-29 human colon carcinoma cells to determine the structural requirements for the cross-linking activity of cma, and the role of this activity in the antitumor ... | 1987 | 3664494 |
| non-random distribution of alu-family repeats in human chromosomes. | a phage lambda recombinant clone containing at least 8 alu-family repeats (afrs) has been isolated from a human genomic library, and dna from the phage was used as a probe for in situ hybridization on g-banded human metaphase chromosomes of healthy donors and leukemic patients. some chromosome bands show prominent clusters of silver grains in all individuals examined: 1p34, 1q23, 2q21-22, 10p14, 11p14, 10q21 and 11q14. the data suggest non-random distribution of afrs in the human genome. | 1987 | 3670286 |
| assembly in vitro of nuclei active in nuclear protein transport: atp is required for nucleoplasmin accumulation. | dna (from bacteriophage lambda or xenopus) is assembled into nucleus-like structures when mixed with an extract from xenopus eggs. electron microscopy shows that these in vitro-reconstituted nuclei possess complete double membranes; some, but not all, nuclei have pore complexes. extracts depleted of their endogenous atp (by addition of atpases) cannot assemble nuclear envelopes visible by phase-contrast microscopy. once synthetic nuclei are assembled, however, they are stable when atp is subsequ ... | 1986 | 3709518 |
| isolation and sequence of cdna clones coding for the precursor to the gamma subunit of mouse muscle nicotinic acetylcholine receptor. | cdna libraries have been constructed in plasmid (pbr322) and bacteriophage lambda gammagt10) vectors with poly (a+) rna isolated from the nonfusing mouse muscle cell line bc3h-1. the libraries were screened with a restriction fragment derived from a genomic clone coding for a human acetylcholine receptor gamma subunit. several clones were obtained whose cdna inserts possessed nucleotide and deduced amino acid sequence homology with acetylcholine receptor gamma subunits from torpedo californica, ... | 1986 | 3755765 |
| molecular characterization of the purity of seven human chromosome-specific dna libraries. | we have characterized at the molecular level seven chromosome-specific libraries constructed in phage lambda charon 21a from flow-sorted human chromosomes. the purity of libraries prepared from chromosomes sorted from hamster x human cells was estimated by species-specific hybridization and ranged from 48% to 83% of clones containing human inserts. among libraries of chromosomes from human cells, mass screenings were made for repetitive sequences and 20 clones from the #18 and #20 libraries were ... | 1986 | 3780320 |
| novel dna-protein complex and a large dna in sle cryoprecipitates. | agarose gel electrophoresis of cryoprecipitates from systemic lupus erythematosus (sle) patients revealed the presence of a slowly migrating dnase i-sensitive dna species at the top of the gel. upon deproteinization, electrophoretic migration was modified favouring the migration of a 17.5 kb dna fragment. mixing experiments adding human serum or plasma to a lambda phage dna digest revealed a dna-protein interaction shown by an accumulation of high mol. wt polynucleotide at the top of the gels, a ... | 1986 | 3802574 |
| isolation of a human tissue-type plasminogen-activator genomic dna clone and its expression in mouse l cells. | we have isolated a cdna clone corresponding to a substantial portion of the human tissue-type plasminogen activator (t-pa) protein. it encodes almost all of the protein b chain and part of the 3' untranslated region. we have used this clone to screen bacteriophage lambda and cosmid libraries of human genomic dna. several related genomic clones were isolated. one of these, a cosmid clone, carried approx. 40 kb of human dna. mapping experiments indicate that the region containing the protein-codin ... | 1985 | 3839198 |
| nucleotide sequence of a gene from chromosome 1d of wheat encoding a hmw-glutenin subunit. | a high molecular weight glutenin gene in hexaploid wheat has been isolated by cloning in bacteriophage lambda and characterized. the gene corresponds to polypeptide 12 encoded by chromosome 1d in the variety "chinese spring". the coding sequence predicted contains seven cysteine residues six of which flank a central repetitive region comprising more than 70% of the polypeptide. these findings are related to the role of high molecular weight subunits in the viscoelastic theory of gluten structure ... | 1985 | 3840588 |
| molecular cloning of the complementary dna for human tumor necrosis factor. | tumor necrosis factor (tnf) is a soluble protein that causes damage to tumor cells but has no effect on normal cells. human tnf was purified to apparent homogeneity as a 17.3-kilodalton protein from hl-60 leukemia cells and showed cytotoxic and cytostatic activities against various human tumor cell lines. the amino acid sequence was determined for the amino terminal end of the purified protein, and oligodeoxyribonucleotide probes were synthesized on the basis of this sequence. complementary dna ... | 1985 | 3856324 |
| a dimer of arac protein contacts three adjacent major groove regions of the arai dna site. | contact sites of arac protein to the regulatory site arai of the escherichia coli arabad operon have been determined by the chemical-interference technique. dna fragments were chemically modified an average of once per molecule, and fragments that no longer bound arac were separated by gel electrophoresis from the dna fragments still able to bind the protein. the contact sites were then determined by comparing the positions of modifications in the two dna samples. strong contacts were found with ... | 1985 | 3858809 |
| structural organization of the dr subregion of the human major histocompatibility complex. | two clusters of overlapping cosmid and lambda phage clones comprising 205 kilobases (kb) have been isolated from the dr subregion of the human major histocompatibility complex from a dr4 haplotype. a single dr alpha and three dr beta genes were identified. in one cluster (135 kb), the dr alpha gene is 90 kb distant from the dr beta gene encoding a molecule that carries the mt3 serological specificity. in the second cluster (70 kb), the dr beta gene determining the dr4 specificity is located 22 k ... | 1985 | 3860851 |
| 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 |
| the propeptide of rat bone gamma-carboxyglutamic acid protein shares homology with other vitamin k-dependent protein precursors. | the molecular cloning of bone gamma-carboxyglutamic acid (gla) protein (bgp; osteocalcin) was accomplished by constructing a phage lambda gt11 cdna library from the rat osteosarcoma cell line ros 17/2 and screening this library with antibodies raised against bgp from rat bone. by sequencing several cloned cdnas, we have established a 489-base-pair sequence that predicts a mature bgp of 50 amino acid residues with an nh2-terminal extension of 49 residues. the leader peptide consists of a hydropho ... | 1985 | 3875856 |
| repressor for the sn-glycerol-3-phosphate regulon of escherichia coli k-12: cloning of the glpr gene and identification of its product. | the glpr gene encoding the repressor for the glp regulon of escherichia coli was cloned from a library of hindiii dna fragments established in bacteriophage lambda. phages harboring glpr were isolated by selection for sn-glycerol-3-phosphate dehydrogenase function encoded by glpd, which is adjacent to glpr on the e. coli linkage map. restriction endonuclease analysis and recloning of dna fragments localized glpr to a 3-kilobase-pair ecori-sali segment of dna. strains exhibiting constitutive expr ... | 1985 | 3881401 |
| repair of nonreplicating uv-irradiated dna: cooperative dark repair by escherichia coli uvr and phr functions. | the system previously used to study recombination of nonreplicating uv-irradiated phage lambda dna was adapted to study uv repair. irradiated phages infected undamaged homoimmune lysogens. pyrimidine dimer content (by treatment with micrococcus luteus uv endonuclease and alkaline sucrose sedimentation) and a biological activity endpoint (infectivity in transfection of uvrb reca recb spheroplasts) were followed. unless room light was excluded during dna extraction procedures, photoreactivation (p ... | 1985 | 3881404 |
| molecular cloning and expression of chlamydia trachomatis major outer membrane protein antigens in escherichia coli. | dna obtained from chlamydia trachomatis (serovar l2) was partially digested with dnase i and inserted into the beta-galactosidase gene of bacteriophage lambda gt11. seven recombinants were selected that produced immunoreactive fusion proteins which were detected with anti-c. trachomatis rabbit serum. one recombinant, designated lambda gt11/l2/33, reacted with various monoclonal antibodies that recognize species-, subspecies-, and type-specific determinants on the chlamydial major outer membrane ... | 1985 | 3882565 |
| replication-competent moloney murine leukemia virus carrying a bacterial suppressor trna gene: selective cloning of proviral and flanking host sequences. | a bacterial suppressor trna gene was introduced into the long terminal repeat of the moloney murine leukemia virus (mo-mulv) proviral genome to construct a retrovirus that allows easy cloning of the provirus with flanking host sequences. a replication competent virus, mo-mulv sup containing a trna amber suppressor gene, was derived that replicates to high titers in tissue culture cells and stably transduces the bacterial gene. the recombinant virus can efficiently replicate in vivo when microinj ... | 1985 | 3883352 |
| detection and characterization of a mouse alpha-spectrin cdna clone by its expression in escherichia coli. | a cloned segment of mouse alpha-spectrin mrna has been identified by immunological techniques. double-stranded cdna derived from spleens of anemic mice was introduced into a bacterial expression vector, puc, and transformed escherichia coli colonies were screened by using an antiserum to erythrocyte membrane ghost proteins. of 17 positive colonies, 2 bound antibody to mouse spectrin, and these 2 colonies contained 750-base-pair inserts that cross-hybridized. transfer of the 750-base-pair insert ... | 1985 | 3883358 |
| cloning and genetic analysis of serotype 5 m protein determinant of group a streptococci: evidence for multiple copies of the m5 determinant in the streptococcus pyogenes genome. | a gene bank of group a streptococcus strain manfredo (m protein serotype 5) was constructed with a bacteriophage lambda vector-escherichia coli k-12 host system and screened by immunoblotting hybrid phage plaques with antisera raised to purified pep m5 (serotype 5 m protein fragment released from the streptococcal cell surface by pepsin). hybrid phage expressing m5 antigen (lambda m5) were detected in the gene bank at an unexpectedly high frequency. the cloned streptococcal dna sequences from on ... | 1985 | 3884510 |
| cloning and nucleotide sequencing of the genes for ribosomal proteins s9 (rpsi) and l13 (rplm) of escherichia coli. | the genes for the ribosomal proteins s9 (rpsi) and l13 (rplm) of escherichia coli have been cloned into a lambda phage vector termed l47.1. the two genes were identified by infecting uv-light irradiated cells with the resultant phages and analyzing the protein products by two-dimensional gel electrophoresis. suitable dna fragments of the isolate were cloned subsequently into m13 phage vectors and their nucleotide sequence was determined by the dideoxy method. it is evident that the two genes for ... | 1985 | 3884974 |
| purification and properties of the dnaj replication protein of escherichia coli. | the escherichia coli dnaj gene was originally discovered because mutations in it blocked bacteriophage lambda dna replication. some of these mutations were subsequently shown to interfere with bacterial growth at high temperature, suggesting that dnaj is an essential protein for the host as well. the first step in purifying the dnaj protein was to overproduce it at least 50-fold by subcloning its gene into the pmob45 runaway plasmid. the second step was the development of an in vitro system to a ... | 1985 | 3889001 |
| a protein structure from nuclear magnetic resonance data. lac repressor headpiece. | a procedure is described to determine the three-dimensional structure of biomolecules from nuclear magnetic resonance data. this procedure combines model building with a restrained molecular dynamics algorithm, in which distance information from nuclear overhauser effects is incorporated in the form of pseudo potentials. the method has been applied to the n-terminal dna-binding domain or headpiece (amino acid residues 1 to 51) of the lac repressor from escherichia coli, for which no crystal stru ... | 1985 | 3889346 |
| optimizing the expression in e. coli of a synthetic gene encoding somatomedin-c (igf-i). | double-stranded dna encoding the human hormone somatomedin-c (smc) has been synthesized. this synthetic gene has been inserted into a plasmid bearing the strong leftward promoter (pl) of bacteriophage lambda and expressed in e. coli. codons for the n-terminal region of smc which maximized the hormone's synthesis were chosen in an smc-lac z fusion assay. the amounts of smc accumulated in e. coli were influenced by mutations at two chromosomal loci, lon and htpr. | 1985 | 3889844 |
| isolation of recombinant cdnas encoding chicken erythroid delta-aminolevulinate synthase. | we report the isolation of cdna clones encoding delta-aminolevulinate synthase (ala synthase; ec 2.3.1.37), the first enzyme in the heme biosynthetic pathway in animal cells. the gene was isolated from a chicken erythroid cdna library prepared in the bacteriophage lambda fusion/expression vector gt11, using rabbit antibody raised against the relatively abundant chicken liver enzyme. the chicken liver and red cell ala synthase isozymes share substantial crossreactivity to the antibody, thereby al ... | 1985 | 3889912 |
| envelope protein changes, autoagglutination, sensitivity to hydrophobic agents and a conditional division lesion in escherichia coli strains carrying colv virulence plasmids. | the presence of the virulence plasmids colv,i-k94 or colv-k30 in escherichia coli produces a number of cell membrane and envelope changes. the most striking of these are (1) the presence of the 33k vmpa outer membrane protein and (2) the colv-associated occurrence of autoagglutination. the vmpa protein is a plasmid-encoded outer membrane protein which is synthesized from a larger precursor. it is distinct from the chromosomally-encoded ompa protein but resembles it in a few respects. the vmpa pr ... | 1985 | 3890691 |
| overproduction of staphylokinase in escherichia coli and its characterization. | a recombinant plasmid which directs the overproduction in escherichia coli of staphylokinase from staphylococcus aureus has been constructed by placing the staphylokinase gene, sak, under the control of bacteriophage lambda pr promoter in the plasmid. when an e. coli strain having the plasmid was induced, the staphylokinase activity in the periplasmic fraction increased about 60-fold and the 15.5-kda protein corresponding to the mature form reached about 25% of the periplasmic proteins. at the s ... | 1985 | 3891339 |
| molecular cloning and a stable amplification of the dna molecules heavily methylated at cpg sequences using a new e. coli cell system (gc3). | fish lymphocystis disease virus (fldv) dna is heavily methylated in cpg sequences and therefore the amplification of recombinant dna of fldv inserted into a bacterial plasmid vector and/or a bacteriophage system poses severe problems. the problem was solved using a newly constructed and developed bacterial system (e. coli gc-3 system), which allowed the amplification of foreign dna material heavily methylated at cpg sequences. e. coli gc-3 (gc-1, rifd) is derived from e. coli gc-1 (k 12, hsd-, m ... | 1985 | 3891463 |
| expression of t4 early genes 62, 44, 45 and 46 in the lambda-t4 recombinant phage lambda 806-17. | a lambda-t4 recombinant phage, lambda 806-17, which carries the t4 early genes 62, 44, 45 and 46, was studied inside a homoimmune lysogen. under such conditions, gene expression from the lambda promoters is represented. results showed extensive expression of gene 46, and significant expression of genes 62 and 45. the expression of these early t4 genes is presumed to depend on t4 promoters included in the cloned fragment. a new promoter proximal to gene 46 is implicated. the results also indicate ... | 1985 | 3891913 |
| demonstration of a bacteriophage receptor site on the escherichia coli k12 outer-membrane protein ompc by the use of a protease. | the escherichia coli k12 outer-membrane proteins ompa, ompc, ompf, phoe, and lamb (all of transmembrane nature) can serve as phage receptors. we have shown previously that one ompa-specific phage, ox2, can give rise to the host range mutants ox2h10 and ox2h12, with the latter being derived from the former [morona, r. & henning, u. (1984) j. bacteriol. 159, 579-582]. unlike ox2, both host range phages can use the ompa and ompc proteins as receptors and ox2h12 is better adapted to the ompc protein ... | 1985 | 3894021 |
| [generation of deletions in the region of the crp gene on the escherichia coli chromosome]. | deletions in the argd, crp, cysg genes (73-74 min of the escherichia coli genetic map) were obtained by heat induction of the phage lambda c1857 b221 rex::tn5 integrated previously into the cysg gene by homologous recombination in the cysg::tn5 mutant. properties of the deletions obtained suggest the gene order: argd-crp-cysg. | 1985 | 3896929 |
| fipb and fipc: two bacterial loci required for morphogenesis of the filamentous bacteriophage f1. | we describe the identification of two mutations in bacterial genes, designated as fipb and fipc, which resulted in temperature-sensitive morphogenesis of bacteriophage f1. these mutations mapped at separate loci but had to be present simultaneously to block f1 production at 41.5 degrees c. one mutation defined the locus fipb at 85.3 min on the escherichia coli linkage map; the other defined the locus fipc, which mapped very close to rpsl at 73 min. since these mutations did not appear to affect ... | 1985 | 3897199 |
| translational efficiency of the escherichia coli adenylate cyclase gene: mutating the uug initiation codon to gug or aug results in increased gene expression. | roy et al. [roy, a., haziza, c. & danchin, a. (1983) embo j. 2, 791-797] established that translation of escherichia coli adenylate cyclase initiates at a uug codon, and they suggested this might decrease the efficiency of translation. we investigated the effect of varying the initiation codon on the expression of the adenylate cyclase (cya) gene. using oligonucleotide-directed mutagenesis, we changed the uug initiation codon to gug and the more common initiator aug and assayed for cya gene expr ... | 1985 | 3898067 |
| cloning, expression in escherichia coli, and reconstitution of human myoglobin. | a full-length cdna clone for human myoglobin has been isolated from a human skeletal muscle cdna library. the clone as isolated has a cdna insert approximately one kilobase long and has 5' and 3' untranslated regions of approximately 80 and 530 base pairs, respectively. the sequence of the translated region corresponds exactly to that predicted for human myoglobin. the cdna was expressed in high yield in escherichia coli as a fusion protein consisting of the first 31 amino acids of the phage lam ... | 1985 | 3898068 |
| molecular cloning of staphylococcal enterotoxin b gene in escherichia coli and staphylococcus aureus. | we have cloned the staphylococcus aureus entb gene in escherichia coli, using pbr322 as the vector plasmid; however, no detectable staphylococcal enterotoxin b (seb) was produced by the e. coli clones. when the entb gene was placed downstream from the strong lambda phage promoter, pr, seb was synthesized at readily detectable levels in e. coli. interestingly, mature seb was almost exclusively present in the cytoplasmic fraction. the seb precursor was found associated with the cell membrane. the ... | 1985 | 3898073 |
| evidence for transcription antitermination control of tryptophanase operon expression in escherichia coli k-12. | tryptophanase, encoded by the gene tnaa, is a catabolic enzyme distinct from the enzymes of tryptophan biosynthesis. tryptophanase synthesis is induced by tryptophan and is subject to catabolite repression. we studied the mechanism of tna operon induction. mutants with altered rho factor were partially constitutive for tna expression, implicating rho-dependent transcription termination in the control of tna expression. measurements of mrna synthesis from the transcribed leader region preceeding ... | 1985 | 3902796 |
| genetic mapping and dna sequence analysis of mutations in the pola gene of escherichia coli. | dna polymerase i of escherichia coli provides an excellent model for the study of template-directed enzymatic synthesis of dna because it is a single subunit enzyme, it can be obtained in large quantities and the three-dimensional structure of the polymerizing domain (the klenow fragment) has recently been determined (ollis et al., 1985). one approach to assigning functions to particular portions of the structure is to correlate the altered enzymatic behavior of mutant forms of dna polymerase i ... | 1985 | 3910840 |
| structure-function relationship in allosteric aspartate carbamoyltransferase from escherichia coli. i. primary structure of a pyri gene encoding a modified regulatory subunit. | in a previous article, we have identified a lambda bacteriophage directing the synthesis of a modified aspartate carbamoyltransferase lacking substrate-co-operative interactions and insensitive to the feedback inhibitor ctp. these abnormal properties were ascribed to a mutation in the gene pyri encoding the regulatory polypeptide chain of the enzyme. we now report the sequence of the mutated pyri and show that, during the generation of this pyrbi-bearing phage, six codons from lambda dna have be ... | 1985 | 3912513 |
| effect of amino acid substitutions on conformational stability of a protein. | this paper reviews studies on thermostable proteins from thermophilic bacteria and on mutant proteins of human hemoglobin, tryptophan synthase alpha-subunit of e. coli, t4 phage lysozyme, and phage lambda repressor with respect to the role of the constituting amino acid residues in stabilization of conformation. the stability of a protein is easily affected by single amino acid substitutions, by which the protein undergoes change(s) of one or more of the following: a hydrogen bond, a salt bridge ... | 1985 | 3914832 |
| molecular cloning of the 3-phosphoglycerate kinase (pgk) gene from aspergillus nidulans. | the aspergillus nidulans 3-phosphoglycerate kinase gene (pgk) has been isolated from a phage lambda genomic library, using the equivalent yeast gene as a hybridization probe. the location of the pgk gene within the cloned dna has been physically mapped. the dna sequence of a small region of the putative pgk has been determined and found to code for amino acids corresponding to the n-terminal end of the pgk protein. in contrast to the yeast pgk gene the aspergillus gene contains a 57 base pair in ... | 1985 | 3916724 |
| cloning and characterization of the three enzyme structural genes qutb, qutc and qute from the quinic acid utilization gene cluster in aspergillus nidulans. | heterologous dna probes from the quinic acid gene cluster (qa) in neurospora crassa (schweizer 1981) have been used to isolate the corresponding gene cluster (qut) from aspergillus nidulans cloned in a phage lambda vector. n. crassa probes for each of the three enzyme structural genes in the cluster have been used to identify the corresponding genes within the a. nidulans cloned dna. the three genes are in the same relative sequence [dehydrogenase (1), qa-3 = qutb; dehydratase (3), qa-4 = qutc; ... | 1985 | 3916726 |
| gene encoding human growth hormone-releasing factor precursor: structure, sequence, and chromosomal assignment. | we have isolated and characterized overlapping clones from phage lambda and cosmid human genomic libraries that predict the entire structure of the gene encoding the precursor to human growth hormone-releasing factor. the gene includes five exons spanning 10 kilobase pairs of human genomic dna. there appears to be a segregation of distinct functional regions of the grf precursor and its mrna into the five exons of the gene. the dna sequences of all exons, intron/exon boundaries, and 5' and 3' fl ... | 1985 | 3918305 |
| genetic characterization and partial sequence determination of a treponema pallidum operon expressing two immunogenic membrane proteins in escherichia coli. | a detailed physical and genetic map of a previously cloned 5.5-kilobase segment of treponema pallidum dna is described. this segment expressed two proteins that are cell membrane associated in escherichia coli. the structural genes of these treponemal membrane proteins, tmpa and tmpb, are coordinately expressed, and transcription in e. coli can start from at least two different treponemal promoters. the tmpa and tmpb proteins are the products of in vivo proteolytic cleavage from precursor protei ... | 1985 | 3922944 |
| isolation, characterization, and mapping to chromosome 19 of the human apolipoprotein e gene. | the human apo-e gene has been isolated from a lambda phage library using as a probe the previously reported apo-e cdna clone pe-301. lambda apo-e was mapped and subcloned, and the apo-e gene was completely sequenced. the dna sequence was compared with that of a near full length cdna clone pe-368 and revealed three introns. the first intron was in the region that corresponds to the 5' untranslated region of apo-e mrna. the second intron interrupted the codon specifying amino acid -4 of the apo-e ... | 1985 | 3922972 |
| cloning and expression of the chromosomal immune interferon gene of the rat. | the chromosomal immune interferon gene of the rat (ifn-gamma) was identified by screening a recombinant rat lambda phage library with a human ifn-gamma cdna probe. in contrast to the genes of other rat ifns, this rat ifn-gamma chromosomal gene contains introns and its structural organization closely resembles that of the human and murine ifn-gamma genes. the rat ifn-gamma gene encodes a signal sequence of 19 amino acids followed by the mature ifn-gamma protein of 137 amino acids. the gene was ex ... | 1985 | 3924594 |
| molecular cloning of alpha-amylase genes from drosophila melanogaster. i. clone isolation by use of a mouse probe. | a cloned alpha-amylase cdna sequence from the mouse is homologous to a small set of dna sequences from drosophila melanogaster under appropriate conditions of hybridization. a number of recombinant lambda phage that carry homologous drosophila genomic dna sequences were isolated using the mouse clone as a hybridization probe. putative amylase clones hybridized in situ to one or the other of two distinct sites in polytene chromosome 2r and were assigned to one of two classes, a and b. clone lambd ... | 1985 | 3924727 |
| synthesis and biological activity of 6-substituted mitosene analogues of the mitomycins. | a series of 1-acetoxymitosene analogues, in which the substituent at c-6 was varied, was prepared by total synthesis and screened for activity against p388 leukemia in mice and induction of lambda phage in escherichia coli. among the 6-substituents prepared, none was as effective as the methyl group in conferring biological activity. however, certain n-methylcarbamates were more active than the unsubstituted carbamates. | 1985 | 3925147 |
| synthesis in escherichia coli of the reovirus nonstructural protein sigma ns. | the coding region of reovirus type 3 genomic segment s3, encoding the nonstructural protein sigma ns, was placed under the control of the bacteriophage lambda pl promoter in the escherichia coli expression plasmid prc23 (j.c. lacal, e. santos, v. notario, m. barbacid, s. yamazaki, h.-f. kung, c. seamans, s. mcandrew, and r. crowl, proc. natl. acad. sci. usa 81:5305-5309). derepression of the pl promoter led to the synthesis of a protein of the same molecular weight as sigma ns produced in reovir ... | 1985 | 3932675 |
| characterization of the polypeptide composition of human factor viii:c and the nucleotide sequence and expression of the human kidney cdna. | human coagulation factor viii:c has been purified approximately 5000-fold from commercial preparations with an average activity yield of 35%. proteins of 92 kd and 77-80 kd enriched during purification are precipitated by a human serum polyclonal antibody which inhibits factor viii:c activity. evidence suggests that these polypeptides are linked by a calcium ion bridge. partial amino acid sequence information from these proteins has been obtained from the intact polypeptides and from products of ... | 1985 | 3935400 |
| [control of lysogeny and a genetic map of the temperate phage sm of pseudomonas aeruginosa]. | 76 mutants with impaired ability to lysogenize host cells were isolated in sm phage after mutagenesis using several chemical mutagens. by means of complementation test, these mutants were distributed into two groups, ci and cii. the mutants of the ci group were similar phenotypically to the ci mutants of phage lambda defective in synthesis of repressor. the mutants of the cii group establish and support the lysogenic state in infected cells with very low frequency. temperature-sensitive mutants ... | 1985 | 3935514 |
| n-band proteins of nucleolar organizers: chromosomal mapping, subnucleolar localization and rdna binding. | the ribosomal dna(rdna)-containing chromatin in eukaryotes forms a unique architecture called the "secondary constriction" or "nucleolus organiser region (nor)" on mitotic chromosomes. to gain more insight into non-histone chromosomal proteins (nhcp), termed "n-band proteins", that are specifically associated with the nor in a wide variety of eukaryotes, we attempted to: identify the nhcp responsible for n-band staining; determine their stoichiometry; map them on metaphase chromosomes; determine ... | 1986 | 3948600 |
| molecular cloning of unintegrated closed circular dna of porcine retrovirus. | viral dna of unintegrated closed circular form was isolated from a swine kidney cell line (skl) which was infected with a porcine retrovirus tsukuba-1 (pretv) produced from a swine malignant lymphoma-derived cell line. shimozuma-1 and cloned using a lambda phage vector, charon 21a. one of ten independent clones contained the 8.3 kb dna fragment as an insert, which was thought to be a full length of viral dna molecule carrying a long terminal repeat (ltr) sequence. we have analyzed this insert by ... | 1986 | 3956742 |
| overproduction and purification of the products of bacteriophage t3 genes 18 and 19, two genes involved in dna packaging. | the products of gene 18 (gp18) and gene 19 (gp19) of bacteriophage t3 are noncapsid proteins involved in dna packaging. a restriction fragment containing gene 18 or 19 was cloned into the plasmid vector pnt45 under the control of the inducible leftward promoter (pl) of phage lambda. induction of transcription of gene 18 or 19 by derepression of the pl promoter led to the synthesis of a high level of gp18 or gp19. by using complementation of t3 dna packaging in vitro as an assay, gp18 and gp19 we ... | 1986 | 3962187 |
| polypeptide ligation occurs during post-translational modification of concanavalin a. | lectins are proteins with multivalent carbohydrate-binding sites, which confer the ability to agglutinate. the seeds of legumes are particularly rich in lectins, for example, concanavalin a (con a) comprises up to 15% of the protein in the cotyledons of jack bean (canavalia ensiformis) seeds. the amino acid sequences of con a and several other legume lectins have been partially or fully determined, and comparison of these sequences from different species reveals a circular homology (fig. 1a); re ... | 1985 | 3965973 |
| the morphology of l-fucose, d-mannose specific lectin (sfl 100-2) produced by streptomyces no. 100-2. | the morphology of an l-fucose specific lectin, sel 100-2, from a streptomyces sp. was studied. electron microscopic observation showed that purified sfl 100-2 preparation consisted of particles homogeneous in size. the diameter was 25 nm. the digitized images of these particles had 2-fold rotation symmetry. the sedimentation coefficient (s020,w) was determined to be 20.6s. the particle weight and the stokes radius were calculated to be 8.0 x 10(5) daltons and 94 a, respectively, by three indepen ... | 1985 | 4008461 |
| organization of the ribosomal rna genes in schistosoma mansoni. | the organization of the rrna genes of schistosoma mansoni has been determined by southern blot analysis of genomic dna digested with restriction enzymes, by isolation of the entire repeat on a single fragment of about 11 kilobase pairs from a genomic dna library constructed in bacteriophage lambda and by characterization of three cloned ecori fragments which span the entire repeat. the segments encoding both the large and small rrna subunits have been identified using specific cloned yeast rdna ... | 1985 | 4010707 |
| comparison of controlled pore glass and kieselguhr-polydimethylacrylamide composite as supports for solid-phase synthesis of 23-residue oligodeoxyribonucleotides in milligram amounts. | two 23-residue oligodeoxyribonucleotides, corresponding to both strands of a dna duplex at the or3 site of bacteriophage lambda, have been synthesized in good yields and in milligram quantities by a solid-phase phosphotriester method using two different supports, kieselguhr-polydimethylacrylamide composite and controlled pore glass. rapid purification was possible using high-performance liquid chromatography on radial compression ion-exchange columns. the results and utility of the two supports ... | 1985 | 4025825 |
| agarose gel electrophoretic evidence for domains of nuclear dna linked with bonds cleavable with sulfhydryl molecules. | complexes of intact nuclear dna with proteins undissociable by 2.0 m nacl and nonionic detergents were analyzed by agarose gel electrophoresis following physical or enzymatic fragmentation. sulfhydryl molecules converted these dnas (but not the bacteriophage lambda dna) into smaller-mr forms. following limited restriction endonuclease digestion of complexes with psti most of the nuclear dna formed a high-molecular-mass band in the 60-110 kbp range. these 60-110 kbp fragments, releasable from the ... | 1985 | 4054300 |
| in vivo transcription of ribosomal rna in relation to the mitotic cycle in physarum polycephalum. | we have investigated the transcription of ribosomal rna in plasmodia of physarum polycephalum by a combination of pulse-labelling with [3h]uridine and rna:dna hybridization. the dna used for the hybridization was a hindiii restriction fragment (cloned in bacteriophage lambda) of physarum ribosomal dna that carries a substantial fraction of the rrna genes, enabling the ribosomal transcripts in the newly synthesized rna to be measured. we found that ribosomal rna constituted about 40% of the pulse ... | 1985 | 4066783 |
| the ribosomal rna genes of locusta migratoria: copy number and evidence for underreplication in a polyploid tissue. | from libraries of locusta migratoria genomic dna in bacteriophage lambda, clones have been isolated that hybridize with one or both of the 18s and 28s components of locust rrna. six clones studied show common patterns of restriction sites in the coding sequences and some include an intron in the 28s region. subcloned sequences from within the 18s and 28s coding regions were used as probes in dot hybridization assays of the rrna gene copy number in dna prepared from several locust tissues and sta ... | 1985 | 4075222 |
| increasing and decreasing protein stability: effects of revertant substitutions on the thermal denaturation of phage lambda repressor. | the thermal denaturations of five revertant lambda repressors containing single amino acid substitutions in their n-terminal domains have been studied by differential scanning calorimetry. two substitutions slightly decrease stability, and the remaining three render the protein more stable than wild type. the gly48----asn and gly48----ser proteins are 4 degrees c more stable than wild type. these two substitutions replace an alpha helical residue, and in each case a poor helix forming residue, g ... | 1985 | 4077930 |
| [genetic recombination and "lex" function in uv reactivation and mutagenesis of phage lambda]. | 1970 | 4098020 | |
| the morphogenesis of bacteriophage lambda. i. purification and characterization of lambda heads and lambda tails. | 1970 | 4099070 | |
| self-assembly of bacteriophage lambda tails. | 1971 | 4106183 | |
| lysis defective mutants of bacteriophage lambda: on the role of the s function in lysis. | 1971 | 4107551 | |
| breakdown of bacteriophage lambda dna in "escherichia coli" k12 "rec." bacteria. | 1971 | 4107886 | |
| isolation and protein composition of normal and petit capsids of bacteriophage lambda. | 1971 | 4108615 | |
| the in situ morphological transition of the bacteriophage lambda tubular head-forms. | 1971 | 4109518 | |
| morphogenesis of bacteriophage phi 80: identification of the cistron 13 product. | an in vitro complementation reaction leading to the assembly of bacteriophage phi80 tails from component proteins is described. tail assembly occurs when a lysate of any mutant in cistron 13 is mixed with a second lysate of a mutant in any of the other cistrons involved in tail formation. lysates of mutants that are blocked in tail formation contain phage heads that can unite with free tails to form infective particles. the rate of the complementation reaction shows little dependence upon temper ... | 1972 | 4110106 |
| the morphogenesis of phage lambda. v. form-determining function of the genes required for the assembly of the head. | 1972 | 4113245 | |
| the capsid structure of bacteriophage lambda. | 1973 | 4125251 | |
| [a bacterial mutation augmenting the frequency of lysogenation by lambda phage]. | 1973 | 4128299 | |
| [study of the characteristics of lambda phage dna transfection in e. coli cells treated with cacl2. the dependence of transfection on the phase of culture growth. the effect of actinomycin on transfection]. | 1973 | 4132141 | |
| an escherichia coli mutant which inhibits the injection of phage lambda dna. | 1974 | 4132241 | |
| the alleviation of host-controlled restriction of unmodified phages by functions of bacteriophage lambda. | 1973 | 4133568 | |
| letter: capsid structure of bacteriophage lambda. | 1974 | 4141737 | |
| [the replication of "precocious" defective mutants of lambda phage]. | 1967 | 4167022 | |
| [action of ultraviolet rays on bacteriophage lambda]. | 1968 | 4174324 | |
| [a mutant of bacteriophage lambda, resistant to acridine orange]. | 1966 | 4178198 | |
| the lysozyme of bacteriophage lambda. ii. amino acid and end group analysis. | 1969 | 4181160 | |
| isolation of the bacteriophage lambda receptor from escherichia coli. | a factor which inactivates the phage lambda can be extracted from escherichia coli. this factor is a protein and is located in the outer membrane of the bacterial envelope. it is found in extracts of strains which are sensitive to phage lambda, but not in extracts of strains specifically resistant to this phage. we conclude that this factor is the lambda receptor, responsible for the specific adsorption of the phage lambda to e. coli cells. a partial purification of the lambda receptor is descri ... | 1973 | 4201774 |
| a spheroplast assay for recombination in bacteriophage lambda dna. | 1974 | 4206975 | |
| mechanism of inhibition of bacillus subtilis dna polymerase 3 by the arylhydrazinopyrimidine antimicrobial agents. | arylhydrazinopyrimidines inhibit dna synthesis in bacillus subtilis by promoting formation of a specific, long-lived ternary complex with dna polymerase iii and the template-primer dna. dna polymerase iii contains an associated, single-strand-selective exonuclease which generates 5'-mononucleotides. drug inhibition of the nuclease similarly proceeds through formation of the ternary complex. the ternary complex was isolated by agarose chromatography. like inhibition of the nuclease, optimum forma ... | 1974 | 4213240 |
| viable molecular hybrids of bacteriophage lambda and eukaryotic dna. | a bacteriophage lambda strain has been constructed and a method developed by which dna from potentially any source can be covalently inserted through ecori cohesive ends into the middle of the lambda dna. these hybrid dnas can infect nonrestricting escherichia coli cells and can then propagate as plaque-forming phage. a unique feature of this lambda strain is that extra dna in the middle of its genome is required for plaque formation. a large number of such phages have been produced with e. coli ... | 1974 | 4216019 |