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a cloned tryptophan-synthesis gene from the ascomycete cochliobolus heterostrophus functions in escherichia coli, yeast and aspergillus nidulans.a gene (trp1) in the tryptophan biosynthetic pathway of the fungal plant pathogen cochliobolus heterostrophus was isolated by complementation of an escherichia coli trpf mutant which lacked phosphoribosylanthranilate isomerase (prai) activity. the cloned gene also complemented an e. coli trpc mutant lacking indoleglycerolphosphate synthase (igps) activity, a yeast trp1 mutant missing prai activity and an aspergillus nidulans trpc mutant. it functioned in e. coli and a. nidulans without apparent ...19862941339
the effect of papulacandin b on (1----3)-beta-d-glucan synthetases. a possible relationship between inhibition and enzyme conformation.the antibiotic, papulacandin b, inhibited growth or (1----3)-beta-d-glucan synthetase (or both) in the fungi saccharomyces cerevisiae, hansenula anomala, neurospora crassa, cryptococcus laurentii, schizophyllum commune and wangiella dermatitidis. no effect was observed on achlya ambisexualis. there was no apparent correlation between the inhibition of growth and that of the synthetase. with most of the fungal extracts, the inhibition of glucan synthetase by papulacandin b became less pronounced ...19862942248
biochemical comparison of the neurospora crassa wild type and the temperature-sensitive and leucine-auxotroph mutant leu-5. purification of the cytoplasmic and mitochondrial leucyl-trna synthetases and comparison of the enzymatic activities and the degradation patterns.the cytoplasmic leucyl-trna synthetases of neurospora crassa wild type (grown at 37 degrees c) and mutant (grown at 28 degrees c) were purified approximately 1770-fold and 1440-fold respectively. additional enzyme preparations were carried out with mutant cells grown for 24 h at 28 degrees c and transferred then to 37 degrees c for 10-70 h of growth. the mitochondrial leucyl-trna synthetase of the wild type was purified approximately 722-fold. the mitochondrial mutant enzyme was found only in tr ...19862942398
regulation of fungal cell wall growth: a guanine nucleotide-binding, proteinaceous component required for activity of (1----3)-beta-d-glucan synthase.by treatment with detergent and nacl, particulate (1----3)-beta-d-glucan synthase (ec 2.4.1.34) from hansenula anomala or neurospora crassa was dissociated into a "soluble fraction" and a "membrane fraction." each fraction alone was almost inactive, but enzymatic activity could be reconstituted by mixing the two fractions and adding gtp or one of its analogs. based on their lability to heat and to incubation with trypsin, the activity in both fractions is proteinaceous. the active component in t ...19862942941
compartmental and regulatory mechanisms in the arginine pathways of neurospora crassa and saccharomyces cerevisiae. 19862945985
isolation and partial nucleotide sequence of the laccase gene from neurospora crassa: amino acid sequence homology of the protein to human ceruloplasmin.the laccase (benzenediol:oxygen oxidoreductase, ec 1.10.3.2) gene from neurospora crassa was cloned and part of its nucleotide sequence corresponding to the carboxyl-terminal region of the protein has been determined. the gene was cloned by cdna synthesis with a laccase-specific synthetic deoxyundecanucleotide as primer and poly(a) rna isolated from cycloheximide-treated n. crassa cultures as template. based on the nucleotide sequence of the cdna obtained, a unique 21-mer was synthesized and use ...19862947240
involvement of chla, e, m, and n loci in escherichia coli molybdopterin biosynthesis.all molybdenum enzymes except nitrogenase contain a common molybdenum cofactor, whose organic moiety is a novel pterin called molybdopterin (mpt). to assist in elucidating the biosynthetic pathway of mpt, two mpt-deficient mutants of escherichia coli k-12 were isolated. they lacked activities of the molybdenum enzymes nitrate reductase and formate dehydrogenase, did not reconstitute apo nitrate reductase from a neurospora crassa nit-1 strain, and did not yield form a, a derivative of mpt. by p1 ...19872947896
localization of alkaline phosphatase activity at microbody membranes of neurospora crassa and aspergillus nidulans.hyphal cells of neurospora crassa and aspergillus nidulans, grown in sabouraud glucose broth or in a defined medium with xanthine or its catabolites as the nitrogen source, contained single membrane-bound organelles cytochemically identified as microbodies. modified gomori procedures at the ultrastructural level revealed putative alkaline phosphatase activity sites in thin sections of cells of both species of fungi. microbody membranes displayed electron opaque deposits (lead phosphate) which we ...19862948778
mitochondrial gene urfn of neurospora crassa codes for a long polypeptide with highly repetitive structure.the mitochondrial dna of neurospora crassa contains a long potential gene, designated urfn, which is located immediately downstream from the co1 gene. these two genes are encoded in different reading frames and overlap by 13 codons. urfn is 633 triplets long and terminates at a uag stop codon. its codon usage is atypical for n. crassa mitochondrial exons and introns, and resembles that of the long open reading frame (orf) of the mitochondrial plasmid present in n. crassa strain mauriceville. mul ...19862949084
sequence analysis and transformation by the catabolic 3-dehydroquinase (qute) gene from aspergillus nidulans.the induction of catabolic 3-dehydroquinase by quinic acid in aspergillus nidulans has been shown to involve transcriptional control and yields a single major 0.8 kb mrna. the nucleotide sequence of the catabolic 3-dehydroquinase qute gene has been determined and contains a single uninterrupted open reading frame of 462 bases encoding a 16,505 da protein of 153 residues. comparison with the corresponding qa2 gene of neurospora crassa reveals the absence of 75 nucleotides encoding 25 amino acids ...19862949740
purification and characterization of 3-dehydroquinase from escherichia coli.a procedure has been developed for the purification of 3-dehydroquinase from escherichia coli. homogeneous enzyme with specific activity 163 units/mg of protein was obtained in 19% overall yield. the subunit mr estimated from polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate was 29,000. the native mr, estimated by gel permeation chromatography on sephacryl s-200 (superfine) and on tsk g3000sw, was in the range 52,000-58,000, indicating that the enzyme is dimeric. the ...19862950851
fungal small nuclear ribonucleoproteins share properties with plant and vertebrate u-snrnps.snrnas with properties closely related to those of the major vertebrate u-snrnas are present in the fungi aspergillus nidulans, neurospora crassa and schizosaccharomyces pombe. these rnas possess a tri-methyl guanosine cap structure and a subset cross-hybridizes with human u1 and u2 clones. in the form of snrnps, snrnas from these fungi as well as from saccharomyces cerevisiae and pea plants are immunoprecipitated by human and anti-sm or anti-(u1)rnp autoimmune antibodies. on micro-injection int ...19872953599
purification and characterization of arginase from neurospora crassa.we have purified an enzymatically active form of arginase from a wild-type strain of neurospora crassa to homogeneity. the enzyme has a subunit molecular weight of 38,300 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. the native protein migrated as a hexamer during gel-filtration chromatography with an apparent molecular weight of 266,000. the enzyme exhibited hyperbolic kinetics at ph 9.5 with an apparent km for arginine of 131 mm. antiserum was prepared against the ...19872953715
the neurospora crassa metallothionein gene. regulation of expression and chromosomal location.the promoter region of the neurospora crassa metallothionein gene contains no sequences which are similar to the mammalian or the yeast metal responsive elements (münger, k., germann, u. a., and lerch, k. (1985) embo j. 4, 2665-2668). we therefore studied the regulation of expression of the n. crassa metallothionein gene in response to different metal ions (cu2+, cd2+, zn2+, co2+, and ni2+) by northern analysis. only copper led to the induction of metallothionein mrna. in n. crassa cultures inoc ...19872953720
ornithine decarboxylase from neurospora crassa. purification, characterization, and regulation by inactivation.ornithine decarboxylase, a highly regulated enzyme of the polyamine pathway, was purified 670-fold from mycelia of neurospora crassa that were highly augmented for enzyme activity. the enzyme is significantly different from those reported from three other lower eucaryotic organisms: saccharomyces cerevisiae, physarum polycephalum, and tetrahymena pyriformis. instead, the enzyme closely resembles the enzymes from mammals. the mr = 110,000 enzyme is a dimer of 53,000 da subunits, with a specific a ...19872953728
complementation of area- regulatory gene mutations of aspergillus nidulans by the heterologous regulatory gene nit-2 of neurospora crassa.loss-of-function mutations in the regulatory gene area of aspergillus nidulans prevent the utilization of a wide variety of nitrogen sources. the phenotypes of nit-2 mutants of neurospora crassa suggest that this gene may be analogous to the area gene. transformation has been used to introduce a plasmid containing the nit-2 gene into a. nidulans. the nit-2 gene of neurospora complemented mutations in the area gene, restoring the ability to use a variety of nitrogen sources. this indicated that t ...19872954160
nucleotide sequence and characterization of the pyrf operon of escherichia coli k12.the pyrf gene of escherichia coli k12, which encodes the pyrimidine biosynthetic enzyme orotidine-5'-monophosphate (omp) decarboxylase, is part of an operon that includes a downstream gene designated orff. the orff gene product is a small polypeptide of unknown function. the nucleotide sequence of a 1549-base pair chromosomal fragment containing this operon was determined. an open reading frame capable of encoding the 27-kda omp decarboxylase subunit was identified and shown to be the pyrf struc ...19872956254
a portable signal causing faithful dna methylation de novo in neurospora crassa.methylation of cytosine residues in eukaryotic dna is common, but poorly understood. typically several percent of the cytosines are methylated; however, it is unclear what governs which sequences eventually become modified. neurospora crassa dna containing the "zeta-eta" (zeta-eta) region, which is a region of unusually heavy methylation, was tested for its ability to direct dna methylation de novo. dna stripped of its methylation by propagation in escherichia coli was reintroduced into neurospo ...19872958937
luminescence emission from the cu(i)-thiolate complex in metallothioneins.the luminescence emission properties of cu-metallothioneins (from neurospora crassa, agaricus bisporus and livers of bedlington terriers affected by copper toxicosis) as well as of (cu,zn)-metallothionein from bovine fetal liver are reported. upon excitation in the u.v., these proteins emit a largely red-shifted luminescence with a maximum at 565 nm attributable to the cu(i)-thiolate chromophores of the proteins. differences in the shapes of the spectra and the emission intensity are observed wi ...19872959510
orientation of enzymic domains in tryptophan synthase of neurospora crassa: an immunoblot analysis of trp3 mutant products.extracts of 52 trp3 mutants of neurospora crassa were tested for the presence of serologically cross-reacting material by the method of electrophoretic blot analysis. the test antigen was obtained by excision of lightly stained bands of denatured pure tryptophan synthase after sds-polyacrylamide gel electrophoresis. rabbit antisera raised against this antigen neutralized and precipitated native tryptophan synthase. of the 52 strains, 19 exhibited banding patterns similar to wild type on electrop ...19872959839
molecular genetic analysis of the pyr-4 gene of neurospora crassa.by means of s1 nuclease mapping, one transcription origin and three termini are identified for the pyr-4 gene of neurospora crassa, the same origin being used also by escherichia coli on the cloned gene. translation of the clone in mini-cells gives a 50,000 dalton gene product, the same size as that determined for the neurospora native enzyme. putative caat and tata boxes, and upstream and downstream potential secondary structures, are identified.19872959843
mitochondrial porin of neurospora crassa: cdna cloning, in vitro expression and import into mitochondria.cdna encoding porin of neurospora crassa, the major protein component of the outer mitochondrial membrane, was isolated and the nucleotide sequence was determined. the deduced protein sequence consists of 283 amino acids (29,979 daltons) and shows sequence homology of around 43% to yeast porin; however, no significant homology to bacterial porins was apparent. according to secondary structure predictions, mitochondrial porin consists mainly of membrane-spanning sided beta-sheets. porin was effic ...19872960519
role of siderophores in iron storage in spores of neurospora crassa and aspergillus ochraceus.spores of neurospora crassa 74a are lacking in ferritinlike iron pools, as demonstrated by mössbauer spectroscopic analysis. the cyclic hexapeptide siderophore ferricrocin constituted 47% of the total iron content in spores. after germination and growth, the ferricrocin iron pool disappeared, indicating that the metal was utilized. in spores of aspergillus ochraceus, 74% of the total iron content was bound by ferrichrome-type siderophores. siderophores may function as iron storage forms in funga ...19872960664
secretory protein translocation in a neurospora crassa in vitro system. hydrolysis of a nucleoside triphosphate is required for posttranslational translocation.an in vitro translocation system has been reconstituted with subcellular fractions from the cell wall-less mutant of neurospora crassa (fz;sg;os-1). prepro alpha factor and invertase, secretory proteins from yeast, were faithfully translocated and glycosylated by neurospora microsomes when presence cotranslationally in the neurospora translation system. when presence cotranslationally in the neurospora translation system, microsomes from canine pancreas(crm) could also translocate and glycosylat ...19872960680
isolation and characterization of a neurospora crassa ribosomal protein gene homologous to cyh2 of yeast.we have isolated and characterized a neurospora crassa gene homologous to the yeast cyh2 gene encoding l29, a cycloheximide sensitivity-conferring protein of the cytoplasmic ribosome. the cloned neurospora gene was isolated by cross-hybridization to cyh2. it was sequenced from both cdna and genomic clones. the coding region is interrupted by seven intervening sequences. its deduced amino acid sequence shows 70% homology to that of yeast ribosomal protein l29 and 60% homology to that of mammalian ...19872960953
effects of mammalian insulin on metabolism, growth, and morphology of a wall-less strain of neurospora crassa.addition of mammalian insulin to a nutritionally rich, chemically defined culture medium affects neurospora crassa "slime" (wall-less) cells, as indicated by enhancement of growth, extension of viability at the stationary phase of growth, alteration of morphology, and stimulation of glucose oxidation. bovine, porcine, and recombinant human insulin had similar effects on growth and morphology, while proinsulin, reduced insulin, and several other proteins were inactive. insulin added in the presen ...19882962851
stimulation by mammalian insulin of glycogen metabolism in a wall-less strain of neurospora crassa.addition of bovine insulin to cells of the wall-less variant fgsc4761 of neurospora crassa ("slime") produced several significant effects on glycogen metabolism. 1) intracellular levels of the glycogen precursor udp-glucose decreased 17-18% (p less than 0.01) within 30 min of insulin addition. 2) cells grown with insulin possessed 40% more glycogen than did control cells. 3) the incorporation of 14c-labeled glucose into glycogen increased 41% after 30-min treatment with 100 nm bovine insulin (p ...19882962852
neurospora tryptophan synthase. characterization of the pyridoxal phosphate binding site.tryptophan synthase, which catalyzes the final step of tryptophan biosynthesis, is a multifunctional protein that requires pyridoxal phosphate for two of its three distinct enzyme activities. tryptophan synthase from neurospora crassa, a homodimer of two 75-kda subunits, was shown to bind 1 mol of pyridoxal phosphate/mol of subunit with a calculated dissociation constant for pyridoxal phosphate of 1.1 microm. the spectral properties of the holoenzyme, apoenzyme, and reconstituted holoenzyme were ...19882966157
damage-resistant dna synthesis in eukaryotes.the molecular basis of sensitivity of ionizing radiation and other damaging agents is not clearly defined in eukaryotes. while a large number of mutants have been described only a few have been demonstrated to have a defect in the repair of damage to dna. an interesting characteristic of a sub-group of these mutants, in different species extending throughout the phylogenetic scale, is the presence of damage-resistant dna synthesis. this phenomenon is observed in cells from individuals with the g ...19882966294
effect of the homokaryotic state of the uvs-2 allele in neurospora crassa on formaldehyde-induced killing and ad-3 mutation.formaldehyde was tested for its killing and mutagenic activities in the ad-3 forward-mutation test in neurospora crassa. the test was conducted in 3 two-component heterokaryons (dikaryons) of n. crassa in order to determine the effect of the uvs-2 allele, which causes a defect in nucleotide excision repair, on formaldehyde-induced killing and the induction of ad-3 mutants. these dikaryons were homokaryotic for uvs-2+ (h-12), homokaryotic for usv-2 (h-59), and heterokaryotic for uvs-2 (h-71). for ...19882966296
expression of heat shock genes of neurospora crassa: effect of hyperthermia and other stresses on mrna levels.neurospora crassa mycelium was heat shocked for intervals varying from 15-180 min. heat shock mrna was monitored by hybridization of northern blots with the drosophila hsp-70 gene probe and an inducible member of the yeast hsp-70 gene family, yg100. a 2.7 kilobase (kb) transcript, with homology to these two probes, was detected in cultures shocked for 15 min; its levels increased up to 60-90 min and declined thereafter. sodium arsenite, too, induced the synthesis of this transcript. an additiona ...19882967081
regulation of synthesis and secretion of acid and alkaline phosphatases in neurospora crassa.we show that n. crassa represses the production of acid phosphatase at ph higher than 8.0, irrespective of the carbon source used, whereas production was stimulated by sucrose at slightly acidic ph. the same profile of acid phosphatase production was observed in the pho-2a, pho-3a, nuc-1a, nuc-2a and pregc mutant strains. we also show that acid phosphatase synthesized by the pregc mutant strain grown on high phosphate medium has pronounced differences when compared to the enzyme synthesized by t ...19872967123
plasmid recovery from transformants and the isolation of chromosomal dna segments improving plasmid replication in neurospora crassa.the efficient recovery of plasmid dna from neurospora crassa transformants is described. lithium acetate-treated spores were transformed with plasmid dna and grown in mass in liquid culture. the resulting mycelial growth was harvested and plasmid dna was extracted and used to transform e. coli to ampicillin resistance. although at low frequency, routine recovery of plasmid psd3 which carries the neurospora qa-2+ gene and pbr322 sequences has been demonstrated. about 10% of the recovered plasmids ...19852967124
the cross-pathway control gene of neurospora crassa, cpc-1, encodes a protein similar to gcn4 of yeast and the dna-binding domain of the oncogene v-jun-encoded protein.expression of the gene cpc-1 is required for cross-pathway-mediated regulation of amino acid-biosynthetic genes in neurospora crassa. we have cloned cpc-1 and present an analysis of its structure and regulation. the cpc-1-encoded transcript contains three open reading frames, two of which are located in the 720-nucleotide leader segment preceding the cpc-1 coding region. the two leader open reading frames, if translated, would produce peptides 20 and 41 residues in length. the deduced amino acid ...19882967496
cloning of methylated transforming dna from neurospora crassa in escherichia coli.an arg-2 mutant of neurospora crassa was transformed to prototrophy with a pbr322-n. crassa genomic dna library. repeated attempts to recover the integrated transforming dna or segments thereof by digestion, ligation, and transformation of escherichia coli, with selection for the plasmid marker ampicillin resistance, were unsuccessful. analyses of a n. crassa transformant demonstrated that the introduced dna was heavily methylated at cytosine residues. this methylation was shown to be responsibl ...19882968501
induction of multiple germ tubes in neurospora crassa by antitubulin agents.the antitubulin fungicide benomyl suppressed the linear growth of neurospora crassa wild type strain st. lawrence 74 at micromolar concentrations. the rate of germination of macroconidia was not affected. macroconidia exposed to 1.7 microm benomyl for 5 h formed multiple germ tubes. when germlings incubated for 4 h were exposed to 1.7 microm benomyl for 3 h, their germ tube stopped growing, swelled and emitted several branches. normal linear growth was restored after removal of the fungicide. li ...19882969337
the overexpression, purification and complete amino acid sequence of chorismate synthase from escherichia coli k12 and its comparison with the enzyme from neurospora crassa.the enzyme chorismate synthase was purified in milligram quantities from an overproducing strain of escherichia coli. the amino acid sequence was deduced from the nucleotide sequence of the aroc gene and confirmed by determining the n-terminal amino acid sequence of the purified enzyme. the complete polypeptide chain consists of 357 amino acid residues and has a calculated subunit mr of 38,183. cross-linking and gel-filtration experiments show that the enzyme is tetrameric. an improved purificat ...19882969724
a neurospora crassa heat-shocked cell lysate translates homologous and heterologous messenger rna efficiently, without preference for heat shock messages.cell-free protein synthesis systems were prepared from normally-grown (n-lysate) and heat-shocked (hs-lysate) neurospora crassa mycelium. although both lysates translated homologous mrna, the hs-lysate was more active, yielding a higher incorporation of [35s]-methionine into hot tca-insoluble material and a vastly superior protein synthesis profile. the optimal temperature for translation by both lysates was 21 degrees c; the hs-lysate did not translate heat-shock mrna preferentially at any temp ...19882969781
a specific insulin receptor and tyrosine kinase activity in the membranes of neurospora crassa.cells of the wall-less ("slime") strain of neurospora crassa possess specific high affinity insulin binding sites on their cell surface. 125i-labeled bound insulin was not displaced from these cells by insulin-like growth factor ii (igf-ii), and was only weakly displaced by igf-i and proinsulin. cross-linking of 125i-labeled insulin with n. crassa cells using disuccinimidyl suberate resulted in the labeling of a single band of ca. 67 kda m.w. on a polyacrylamide gel. two proteins of ca. 66 and 5 ...19882970849
x-ray-induced specific locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa. i. modification of the heterozygous effects of multilocus deletions covering the ad-3a or ad-3b loci.the basis for the reduced growth rates of heterokaryons between strains carrying nonallelic combinations of gene/point mutations (ad-3r) and multilocus deletion mutations (ad-3ir) has been investigated by a simple genetic test. the growth rates of forced 2-component heterokaryons (dikaryons) between multilocus deletion mutations were compared with forced 3-component heterokaryons (trikaryons) containing an ad-3ar ad-3br double mutant as their third component. since the third component has no gen ...19882971138
cyclosporin a-binding protein (cyclophilin) of neurospora crassa. one gene codes for both the cytosolic and mitochondrial forms.cyclophilin (cyclosporin a-binding protein) has a dual localization in the mitochondria and in the cytosol of neurospora crassa. the two forms are encoded by a single gene which is transcribed into mrnas having different lengths and 5' termini (approximately 1 and 0.8 kilobases). the shorter mrna specifies the cytosolic protein consisting of 179 amino acids. the longer mrna is translated into a precursor polypeptide with an amino-terminal extension of 44 amino acids which is cleaved in two steps ...19882971658
structure of d-prephenyllactate. a carboxycyclohexadienyl metabolite from neurospora crassa.a novel natural product structurally related to prephenate and arogenate was isolated from a mutant of neurospora crassa. this d-beta-(1-carboxy-4-hydroxy-2,5-cyclohexadiene-1-yl)-lactic acid is herein given the trivial name of d-prephenyllactate. the new metabolite is even more acid labile than is prephenate and is quantitatively converted to phenyllactate at mildly acidic ph. the structure characterization of prephenyllactate was performed using spectroscopic techniques (ultraviolet, 1h nmr, 1 ...19882972718
comparison of the orotidine 5'-monophosphate decarboxylase sequences of eight species.predicted amino acid sequences of the enzyme orotidine 5'-phosphate decarboxylase (ec 4.1.1.23) from eight different organisms are compared. the comparisons are made on the basis of primary structural differences, primary amino acid sequence, hydropathy profiles, and secondary structure predictions. the organisms compared are mus musculus, aspergillus nidulans, neurospora crassa, kluyveromyces lactis, saccharomyces cerevisiae, schizosaccharomyces pombe, escherichia coli, and salmonella typhimuri ...19882974823
sequence analysis of mitochondrial dna from podospora anserina. pervasiveness of a class i intron in three separate genes.a 48 kb region of the 95 kb mitochondrial genome of podospora anserina has been mapped and sequenced (1 kb = 10(3) base-pairs). the dna sequence of the genes for nd2, 3, 4, atpase 6 and urfc are presented here. as in neurospora crassa, the nd2 and 3 genes consist of a unit separated by one taa stop codon. nd3, 4 and atpase 6 are interrupted by class i introns. all three introns are remarkably similar in the c-domain of their secondary structure, sufficient enough to designate them as new subgrou ...19882975708
molecular organisation of the quinic acid utilization (qut) gene cluster in aspergillus nidulans.the functional integrity of the qutb gene (encoding quinate dehydrogenase) has been confirmed by transformation of a qutb mutant strain. the dna sequence of the contiguous genes qutd (quinate permease), qutb and qutg (function unknown) has been determined and analysed, together with that of qute (catabolic 3-dehydroquinase). the qutb sequence shows significant homology with the shikimate dehydrogenase function of the complex arom locus of aspergillus nidulans, and with the qa-3 quinate dehydroge ...19882976880
the amino acid sequence of ribonuclease n1, a guanine-specific ribonuclease from the fungus neurospora crassa.the complete amino acid sequence of ribonuclease n1 (rnase n1), a guanine-specific ribonuclease from a fungus, neurospora crassa, was determined by conventional protein sequencing, using peptide fragments obtained by tryptic digestion of cyanogen bromide-treated rnase n1 and by staphylococcus aureus v8 protease digestion of heat-denatured rnase n1. the results showed that the protein is composed of a single polypeptide chain of 104 amino acid residues cross-linked by two disulfide bonds and has ...19882977130
chemical synthesis and expression of copper metallothionein gene of neurospora crassa.the gene coding for the neurospora crassa copper metallothionein (mt) was synthesized and inserted in the lacz' gene of puc18 plasmid to give the same translational reading frame as the latter gene. the mt-beta-galactosidase fused gene was expressed in escherichia coli to produce a fused protein in which the amino and carboxy termini of mt are linked to the beta-galactosidase through methionine residues. an mt derivative containing an extra homoserine residue at the carboxy terminus was prepared ...19882977386
high-performance liquid chromatography of siderophores from fungi.a reversed-phase hplc separation of iron(iii) chelates of 16 representative fungal siderophores including ferrichromes, coprogens and triacetylfusarinine c was established in order to investigate siderophore production of fungi. for comparison purposes, the widely used bacterial siderophore ferrioxamine b was included. culture filtrates of the fungi penicillium resticulosum, fusarium dimerum, aspergillus fumigatus and neurospora crassa were quantitatively analyzed for the presence of known and u ...19882978959
influence of neurospora endonuclease on trenimon-induced structural chromosomal aberrations and sister-chromatid exchanges in human peripheral lymphocytes and chinese hamster ovary cells.human peripheral lymphocytes and chinese hamster ovary cells were treated in the g1 phase of the cell cycle with the trifunctional alkylating agent trenimon (trn) and post-treated with a single-strand specific endonuclease from neurospora crassa (ne). trn induces chromosomal aberrations of the chromatid type (ca) and sister-chromatid exchanges (sce). ne post-treatment leads to an elevation of the frequencies of ca but not of sces. this indicates that trn induced ca are the result of dna double-s ...19852985980
cloning and characterization of the rdna repeat unit of podospora anserina.dna coding for ribosomal rna in podospora anserina has been cloned and was found as a tandemly repeated 8.3 kb sequence. the cloned rdna was characterized by restriction endonuclease mapping. the location of 5.8s, 18s and 28s rrna coding regions was established by dna-rna hybridization and s1 nuclease mapping. the organization of p. anserina rrna genes is similar to that of neurospora crassa and aspergillus nidulans. the rdna unit does not contain the sequence coding for 5s rna.19852987647
an equilibrium between distorted and undistorted dna in the adult chicken beta a-globin gene.we have used single strand specific nucleases to map dna distortion in the adult chicken beta a-globin gene. we have detected two structures of that kind and have mapped nuclease-cutting sites at one base resolution. one prominent site is centered at -190 relative to the rna capping site and is positioned at the center of a stretch of contiguous c residues. the second site is near the first intron/exon junction (+620) and appears as a series of discrete 1-base-long enzyme-cutting sites. based up ...19852989280
nucleotide sequence of yeast gdh1 encoding nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase.the yeast gdh1 gene encodes nadp-dependent glutamate dehydrogenase. this gene was isolated by complementation of an escherichia coli glutamate auxotroph. nadp-dependent glutamate dehydrogenase was overproduced 6-10-fold in saccharomyces cerevisiae bearing gdh1 on a multicopy plasmid. the nucleotide sequence of the 1362-base pair coding region and 5' and 3' flanking sequences were determined. transcription start sites were located by s1 nuclease mapping. regulation of gdh1 was not maintained when ...19852989290
structure of a neurospora rna polymerase i promoter defined by transcription in vitro with homologous extracts.a neurospora in vitro transcription system has been developed which specifically and efficiently initiates transcription of a cloned neurospora crassa ribosomal rna gene by rna polymerase i. the initiation site of transcription (both in vitro and in vivo) appears to be located about 850 bp from the 5' end of mature 17s rrna. however, the primary rrna transcripts are normally cleaved very rapidly at a site 120-125 nt from the 5' end in vitro and in vivo. the nucleotide sequence surrounding the in ...19852989792
cloning and expression of the fbc operon encoding the fes protein, cytochrome b and cytochrome c1 from the rhodopseudomonas sphaeroides b/c1 complex.the gene for the fes protein of the rhodopseudomonas sphaeroides b/c1 complex was identified by means of cross-hybridization with a segment of the gene encoding the corresponding fes protein of neurospora crassa. plasmids (prsf1-14) containing the cross-hybridizing region, covering in total 13.5 kb of chromosomal dna, were expressed in vitro in a homologous system. one rsf plasmid directed the synthesis of all three main polypeptides of the r. sphaeroides b/c1 complex: the fes protein, cytochrom ...19852990901
comparison of the mitochondrial endonucleases from neurospora crassa and saccharomyces cerevisiae.the endonucleases from neurospora crassa and saccharomyces cerevisiae are not closely related antigenically. they also differ with respect to their activity at ph 8, their degree of hydrophobicity, and their sensitivity to elevated temperatures. however, the two nucleases have similar specific activities, are inhibited by edta, and have nearly identical substrate specificities. since the enzymes also have the same mode of action and intracellular location, these similarities may indicate that th ...19852992732
cloning of the aro cluster gene of neurospora crassa and its expression in escherichia coli.we have constructed a phage, lambda ncl, which comprises a 4.0 kb hindiii insert of neurospora dna into the immunity region of the vector lambda 598. lambda ncl complements the arod6 mutation of e. coli, permitting the formation of galaxy plaques on medium lacking aromatic supplements, and transforms an aro-9 qa-2 neurospora mutant to prototrophy at a low frequency. low levels of 5-dehydroquinate hydrolyase (e.c.4.2.1.10.), with properties unlike those of the catabolic isoenzyme that is coded by ...19852993794
isolation and sequence analysis of a cdna encoding the atp/adp translocator of zea mays l.a cdna complementary to the mrna for the atp/adp translocator of maize (zea mays l.) has been identified by virtue of hybridisation with the homologous gene from yeast. the cloned cdna has been shown by dna sequence analysis to contain an open reading frame of 954bp., which encodes a polypeptide of molecular weight 40,519. this polypeptide exhibits a high degree of homology to the translocator polypeptides of beef heart and neurospora crassa mitochondria.19852994015
excision-amplification of mitochondrial dna during senescence in podospora anserina. dna sequence analysis of three unique "plasmids".during senescence in the filamentous fungus podospora anserina, specific regions of the mitochondrial genome, termed sendna are excised, ligated and amplified. we have cloned in their entirety three such autonomously replicating plasmids, alpha, beta and epsilon sendna. none of these plasmids displayed cross-hybridization nor did we detect any significant dna homology by computer analysis. the complete dna sequence of the 2.5 kb alpha, the 5.5 kb epsilon and about 3.4 kb of the 9.8 kb beta sendn ...19852997455
calmodulin and ca2+-dependent cyclic amp phosphodiesterase activity in trypanosoma cruzi.calmodulin has been purified from trypanosoma cruzi epimastigote forms by ion-exchange chromatography, gel filtration and affinity chromatography on 2-chloro-10-(3-aminopropyl)phenotiazine-sepharose. upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the factor showed a polypeptide band with an apparent molecular weight of 16 000. in addition, cyclic amp phosphodiesterase activity from t. cruzi epimastigote forms was purified by ion-exchange chromatography and affinity chromatograph ...19852999589
correct removal by splicing of a neurospora intron in yeast.processing of intron-containing nuclear messenger rnas in yeast require an internal conserved sequence (ics) element, uacuaac. similar elements (ugcuagac) have been identified in sequences interrupting nuclear genes of the related ascomycete neurospora crassa. to examine the structural splicing requirements in yeast, we constructed hybrid genes containing the intron of the neurospora histone h3 gene and cloned them into high copy number yeast vectors. subsequently we analyzed the rnas transcribe ...19852999703
development of a high-frequency transforming vector for aspergillus nidulans.the pyr4 gene of neurospora crassa, which codes for orotidine-5'-phosphate decarboxylase, is capable of transforming an aspergillus nidulans pyrg mutant by chromosomal integration, despite low homology between the transforming dna and the recipient genome. integration of pfb6, a plasmid carrying pyr4 and capable of replication in escherichia coli, was not observed at the pyrg locus. the efficiency of transformation was considerably enhanced (50-100 fold) by inclusion in the transforming vector o ...19853000883
nuclear genes for cytochrome c oxidase subunits of neurospora crassa. isolation and characterization of cdna clones for subunits iv, v, vi, and possibly vii.we obtained cdna clones for cytochrome oxidase subunits iv, v, vi, and possibly vii by constructing a lambda gt11 library of neurospora crassa cdna and probing it with antiserum directed against neurospora cytochrome oxidase holoenzyme. positive clones were further characterized with antisera directed against individual cytochrome oxidase subunits and subsequently by dna sequencing. the clones for subunits iv and v encode proteins with regions matching the known n-terminal amino acid sequences o ...19863001085
primary structure of tyrosinase from streptomyces glaucescens.the complete amino acid sequence of streptomyces glaucescens tyrosinase is reported. the molecule consists of 273 amino acids and has a mr of 30 900 including two copper atoms. the primary structure was determined by a combination of amino acid and dna sequence analysis. peptide sequence information was derived from the cyanogen bromide, tryptic, and thermolytic fragments of apotyrosinase by automated edman degradation and aminopeptidase m and carboxypeptidase c digestions. the nucleotide sequen ...19853002431
the mitochondrial urf1 gene in neurospora crassa has an intron that contains a novel type of urf.in neurospora crassa, a 2670 base-pair segment of the mitochondrial dna was sequenced including a gene homologous to the mammalian urf1 that was recently shown to encode a subunit of the respiratory chain nadh dehydrogenase complex. urf1 of n. crassa is interrupted by an intron of 1118 base-pairs that divides the protein-coding sequence into two exons of 636 and 480 base-pairs length, respectively. the deduced urf1 polypeptide of 371 residues was aligned with that of other eukaryotes, revealing ...19853003362
characterization of neurospora crassa cyclic amp phosphodiesterase activated by calmodulin.activation of cyclic amp phosphodiesterase i by brain or neurospora calmodulin was studied. the stimulation required micromolar concentrations of ca2+, and it was observed at cyclic amp concentrations between 0.1 and 500 microm. activation was blocked by edta and some neuroleptic drugs such as chlorpromazine and fluphenazine. these drugs inhibit the elongation of n. crassa wild-type aerial hyphae. these results reinforce the evidence towards the recognition of ca2+-calmodulin as one of the syste ...19853004404
cloning and characterization of the multifunctional his-3 gene of neurospora crassa.we have cloned the his-3 gene of neurospora crassa and determined its nucleotide sequence. the gene specifies a protein of 863 amino acids (aa) and contains a 59-bp intron which interrupts aa 800, a proline residue. the 5' end of the his-3 transcript is heterogeneous with major starts 122 and 124 bp upstream from the start codon. there are three possible polyadenylation sites, 119, 120 and 121 bp after the uaa stop codon. the protein shows two regions of homology to the yeast his4 gene which cor ...19853005109
gene cloning in aspergillus nidulans: isolation of the isocitrate lyase gene (acud).an aspergillus nidulans gene library was constructed in a high-frequency transformation vector, pdjb3, based on the neurospora crassa pyr4 gene. this gene library was used to isolate the structural gene for isocitrate lyase (acud) by complementation of a deficiency mutation following transformation of a. nidulans. plasmids rescued in escherichia coli were able to transform five different a. nidulans acud mutants. transformation using plasmids containing the cloned fragment resulted in integratio ...19863010050
cloning of the arg-12 gene of neurospora crassa and regulation of its transcript via cross-pathway amino acid control.the arg-12 locus of neurospora crassa encodes ornithine carbamoyl transferase, which is one of many amino acid synthetic enzymes whose activity is regulated through cross-pathway (or general) amino acid control. we report here the use of probes derived from the functionally equivalent arg-b gene of aspergillus nidulans to identify and clone a 10 kb neurospora dna fragment carrying the arg-12 gene. short neurospora dna probes derived from this fragment were used to identify a 1.5 kb polya+ transc ...19863012277
an immunochemical study of neurospora nucleases.nucleases derived from neurospora crassa mycelia with neutral single-strand (ss) endodeoxyribonuclease activity have been examined by immunochemical techniques and by sodium dodecyl sulfate - dna gel electrophoresis. all of the intracellular nucleases, which have different divalent metal ion requirements, different strand specificities with single- and double-strand dna, different modes of action on dna and rna, and other distinguishing characteristics, are immunochemically related to neurospora ...19863013242
sequence analysis of the pyr-4 (orotidine 5'-p decarboxylase) gene of neurospora crassa.the pyr-4 gene of neurospora crassa encodes orotidine-5' -phosphate decarboxylase, which catalyses the sixth step in the pyrimidine biosynthetic pathway. the complete nucleotide sequence of a 1.8-kb genomic fragment containing the pyr-4 gene has been determined. using transposon mutagenesis, the coding region has been identified, and the amino acid (aa) sequence deduced. comparison of the pyr-4 aa sequence with ura3, the equivalent gene of saccharomyces cerevisiae, showed extensive blocks of hom ...19863019837
alkaline protease deficiency in the cr-1 (crisp) mutant of neurospora crassa.the adenyl cyclase deficient cr-1 mutant of neurospora crassa grew poorly in bovine serum albumin as an alternative and only source of either sulfur, nitrogen or carbon. the low growth of the cr-1 mutant in protein was correlated with limited secretion of extracellular alkaline protease. the defect was specific for the cr-1 mutant and was suppressed by exogenous cyclic amp. cyclic amp relieved protease deficiency under carbon, nitrogen or sulfur limiting conditions to unequal extents. protease s ...19863026533
purification and properties of a single strand-specific endonuclease from mouse cell mitochondria.a nuclease was purified from mitochondria of the mouse plasmacytoma cell line, mcp-11 which acts on single-stranded dna endonucleolytically and appears to have no activity upon native dna. it degrades unordered rna somewhat more effectively than it does dna. the enzyme activity and the major detectable polypeptide migrate to a position corresponding to an mr of 37,400 on denaturing polyacrylamide gels; in its native form the activity has an s value of 4.7, which corresponds to a molecular weight ...19863027656
the primary structure of cytochrome c1 from neurospora crassa.the primary structure of the cytochrome c1 subunit of ubiquinol-cytochrome-c reductase from mitochondria of neurospora crassa was determined by sequencing the cdna of a bank cloned in escherichia coli. from the coding region the sequence of 332 amino acids, corresponding to the molecular mass of 36,496 da, was derived for the precursor protein. the mature protein, the n terminus of which was previously sequenced [tsugita et al. (1979) in cytochrome oxidase (king, t. e. et al., eds) pp. 67-77, el ...19873030747
molecular cloning of a cdna for a human adp/atp carrier which is growth-regulated.we have identified in a human cdna library a clone (hp2f1) whose cognate rna is growth-regulated. the insert has been sequenced and the nucleotide sequence shows a strong homology to the nucleotide sequences of the adp/atp carrier cdna and gene, respectively, isolated from neurospora crassa and saccharomyces cerevisiae. the putative amino acid sequence of hp2f1 shows an 87% homology to the amino acid sequence of the adp/atp carrier from beef heart mitochondria. we conclude that the insert of hp2 ...19873031073
structure and expression of the overlapping nd4l and nd5 genes of neurospora crassa mitochondria.genes homologous to the mammalian mitochondrial nadh dehydrogenase subunit genes nd4l and nd5 were identified in the mitochondrial genome of the filamentous fungus neurospora crassa, and the structure and expression of these genes was examined. the nd4l gene (interrupted by one intervening sequence) potentially encodes an 89 residue long hydrophobic protein that shares about 26% homology (or 41% homology if conservative amino acid substitutions are allowed) with the analogous human mitochondrial ...19873035337
isolation and characterization of the nuclear gene encoding the rieske iron-sulfur protein (rip1) from saccharomyces cerevisiae.the nuclear gene encoding the rieske iron-sulfur protein of the cytochrome bc1 complex of the mitochondrial respiratory chain has been isolated and characterized from saccharomyces cerevisiae. we used a segment of the iron-sulfur protein gene from neurospora crassa (harnisch, u., weiss, h., and sebald, w. (1985) eur. j. biochem. 149, 95-99) to detect the yeast gene by southern analysis. five different but overlapping clones were then isolated by probing a yeast genomic library carried on yep 13 ...19873036836
the yeast aminopeptidase y.a metal-dependent aminopeptidase (ec 3.4.11.-), designated apase y, has been purified to homogeneity by conventional methods. the enzyme is composed of a single polypeptide chain with molecular mass of 102 kilodaltons, estimated by sodium dodecyl sulphate - polyacrylamide gel electrophoresis, with a blocked n-terminal amino acid. it possesses neither endopeptidase nor carboxypeptidase activity and is strongly inhibited by metal-chelating agents, zn2+, and the protein inhibitor from neurospora cr ...19883042113
genetic regulation of the quinic acid utilization (qut) gene cluster in aspergillus nidulans.a large number of quinic acid non-utilizing qut mutants of aspergillus nidulans deficient in the induction of all three quinic acid specific enzymes have been analysed. one class the qutd mutants, are all recessive and are non-inducible at ph 6.5 due to inferred deficiency in a quinate ion permease. two regulatory genes have been identified. the quta gene encodes an activator protein since most quta mutants are recessive and non-inducible although a few fully dominant mutants have been found. th ...19883049934
promoter recognition by escherichia coli rna polymerase. influence of dna structure in the spacer separating the -10 and -35 regions.escherichia coli rna polymerase contacts promoter dna at two regions (the -10 and -35 regions) which are separated by a segment of spacer dna. previously we showed that base substitutions in the spacer dna can affect promoter strength both in vitro and in vivo; these results were interpreted to reflect altered structural properties of the substituted dnas. here we provide experimental support for this interpretation. the pattern of cleavage of the promoters with neurospora crassa endonuclease an ...19883050126
molecular cloning, identification and transcriptional analysis of genes involved in acetate utilization in neurospora crassa.four neurospora crassa genomic clones have been selected as hybridizing much more strongly to labelled mrna isolated from acetate-grown mycelium than to mrna from sucrose-grown mycelium. hybridization of restriction fragments with acetate-specific mrna or cdna has been used to delimit the transcribed region(s) of each clone. the transcription of all four clones is strongly induced by transfer of growing mycelium from sucrose to acetate as sole carbon source. in wild-type mycelium, mrnas correspo ...19883054423
mitotic gene conversion, reciprocal recombination and gene replacement at the bena, beta-tubulin, locus of aspergillus nidulans.we have developed a procedure for determining the rates of mitotic recombination of an interrupted duplication created by integration of transforming plasmid sequences at the bena, beta-tubulin, locus of aspergillus nidulans. transformation of a strain carrying a benomyl-resistant bena allele with plasmid aipgm4, which carries the wild-type bena allele and the pyr4 (orotidine-5'-phosphate decarboxylase) gene of neurospora crassa, creates an interrupted duplication with plasmid sequences flanked ...19883054484
the simple repeat poly(dt-dg).poly(dc-da) common to eukaryotes is absent from eubacteria and archaebacteria and rare in protozoans.genomic dna from a wide variety of prokaryotic and eukaryotic organisms has been assayed for the simple repeat sequence poly(dt-dg).poly(dc-da) by southern blotting and dna slot blot hybridizations. consistent with findings of others, we have found the simple alternating sequence to be present in multiple copies in all organisms in the animal kingdom (e.g., mammals, reptiles, amphibians, fish, crustaceans, insects, jellyfish, nematodes). the tg element was also found in lower eukaryotes (sacchar ...19863127653
evolutionary relationships among copper proteins containing coupled binuclear copper sites.the amino acid sequences of different copper proteins containing coupled binuclear copper centers are compared. hemocyanins from arthropods and molluscs and tyrosinases from three different species were found to share a highly homologous region in the c-terminal parts. this region contains three invariant histidines previously identified as ligands to cu(b) in panulirus interruptus hemocyanin by x-ray crystallography (gaykema et al., nature 309, 23-29 (1984]. in contrast, the ligand environment ...19883136463
stimulation of beta(1----3)glucan synthetase of various fungi by nucleoside triphosphates: generalized regulatory mechanism for cell wall biosynthesis.particulate fractions from the taxonomically diverse fungi achlya ambisexualis, hansenula anomala, neurospora crassa, cryptococcus laurentii, schizophyllum commune, and wangiella dermatitidis were found to catalyze the time-dependent incorporation of glucose from udp-[14c]glucose into a water-insoluble material. the reaction was stimulated by bovine serum albumin. the product was characterized as beta(1----3)glucan on the basis of its resistance to alpha- and beta-amylase and susceptibility to b ...19853156122
protein changes during the asexual cycle of neurospora crassa.a method for synchronizing conidiation and isolating large numbers of cells at discrete stages of conidia development is described. using two-dimensional gel electrophoresis, we analyzed the protein profiles of mycelia, aerial hyphae, and conidia and observed that the concentration of 14 polypeptides increase and 38 decrease during the asexual cycle. twelve polypeptides were present in extracts of aerial hyphae or conidia, but not mycelia, suggesting that they may be conidiation specific. the pr ...19853157863
primary structure of the trpc gene from aspergillus nidulans.we have determined the structure and complete nucleotide sequence of the trifunctional trpc gene from the ascomycetous fungus aspergillus nidulans. results from rna gel blot analyses showed that this gene encodes two size classes of polyribosomal, poly (a)+rnas with approximate lengths of 2,400 and 2,600 nucleotides. s1 nuclease protection studies demonstrated that the distribution into the two size classes is due to selection of alternative sites for polyadenylation. the transcription units con ...19853158796
effect of the homokaryotic or heterokaryotic state of the uvs-2 allele in neurospora crassa on mitomycin c-induced killing and ad-3 mutation.mitomycin c (mc) was tested for its killing and mutagenic activities in the ad-3 forward-mutation test in neurospora crassa. the test was conducted in 4 dikaryons of n. crassa in order to determine the effect of the uvs-2 allele, which causes a defect in nucleotide excision repair, on mc-induced killing and ad-3 mutation. these dikaryons were homokaryotic for uvs-2+ (h-12), homokaryotic for uvs-2 (h-59), and heterokaryotic for uvs-2/uvs-2+ (h-70 and h-71). mc induced killing and ad-3 mutation in ...19853158811
ornithine transcarbamylase from neurospora crassa: purification and properties.ornithine transcarbamylase catalyzes the synthesis of citrulline from carbamyl phosphate and ornithine. this enzyme is involved in the biosynthesis of arginine in many organisms and participates in the urea cycle of mammals. the biosynthetic ornithine transcarbamylase has been purified from the filamentous fungus, neurospora crassa. it was found to be a homotrimer with an apparent subunit molecular weight of 37,000 and a native molecular weight of about 110,000. its catalytic activity has a ph o ...19853159341
primary structure of copper-zinc superoxide dismutase from neurospora crassa.the complete amino acid sequence of copper-zinc superoxide dismutase from neurospora crassa is reported. the subunit consists of 153 amino acids and has a mr of 15,850. the primary structure was determined by automated and manual sequence analysis of peptides obtained by digestions of the carboxymethylated and aminoethylated enzyme with trypsin and thermolysin. the protein is devoid of tryptophan and methionine and displays a free amino terminus. comparison of the amino acid sequence with those ...19853160699
dna sequence and organization of the mitochondrial nd1 gene from podospora anserina: analysis of alternate splice sites.earlier, we reported that the nd1 mitochondrial gene of podospora anserina is mosaic, containing at least three class i introns. we have now completed the sequence of the nd1 gene and have determined that it contains four class i introns of 1,820, 2,631, 2,256 and 2,597 bp with the entire gene complex containing 10,505 bp, only 1,101 of which are exon sequences. introns 1 and 3 appear to be related in that their open reading frames (orfs) exhibit extensive amino acid sequence similarity and like ...19883197134
the gene for rat uncoupling protein: complete sequence, structure of primary transcript and evolutionary relationship between exons.the complete nucleotide sequence of rat uncoupling protein gene has been determined. 4.5 kb of the 5'-flanking region have also been sequenced. the site of transcription start as well as 3'-end extremities were identified. transcription unit spans 8.4 kb and contains 6 exons and 5 introns. uncoupling protein as well as related mitochondrial carriers such as adp/atp carrier and phosphate carrier has a triplicated structure and each repeat of uncoupling protein corresponds to 2 exons. two gene dup ...19883202878
microorganisms associated with mouldiness of dried yam chips and their prevention.the broad objective of this study was to isolate and identify the microorganisms causing mouldiness of stored yam chips and to look for ways of preventing the problem. microorganisms isolated included aspergillus flavus, a. glaucus, a. nidulans, a. niger, a. ochraceous, a. tamarii, a. candidus, penicillium oxalicum, trichoderma longibrachyatum, rhizopus nigricans, cylindrocarpon radicicola, neurospora crassa, botryodiplodia theobromae, bacillus subtilis, bacillus cereus, erwinia carotovora and s ...19883231261
sequencing and overexpression of the escherichia coli aroe gene encoding shikimate dehydrogenase.the escherichia coli aroe gene encoding shikimate dehydrogenase was sequenced. the deduced amino acid sequence was confirmed by n-terminal amino acid sequencing and amino acid analysis of the overproduced protein. the complete polypeptide chain has 272 amino acid residues and has a calculated mr of 29,380. e. coli shikimate dehydrogenase is homologous to the shikimate dehydrogenase domain of the fungal arom multifunctional enzymes and to the catabolic quinate dehydrogenase of neurospora crassa.19883277621
purification and properties of escherichia coli dimethyl sulfoxide reductase, an iron-sulfur molybdoenzyme with broad substrate specificity.dimethyl sulfoxide reductase, a terminal electron transfer enzyme, was purified from anaerobically grown escherichia coli harboring a plasmid which codes for dimethyl sulfoxide reductase. the enzyme was purified to greater than 90% homogeneity from cell envelopes by a three-step purification procedure involving extraction with the detergent triton x-100, chromatofocusing, and deae ion-exchange chromatography. the purified enzyme was composed of three subunits with molecular weights of 82,600, 23 ...19883280546
purification and properties of the major nuclease from mitochondria of saccharomyces cerevisiae.the vast majority of nuclease activity in yeast mitochondria is due to a single polypeptide with an apparent molecular weight of 38,000. the enzyme is located in the mitochondrial inner membrane and requires non-ionic detergents for solubilization and activity. a combination of heparin-agarose and cibacron blue-agarose chromatography was employed to purify the nuclease to approximately 90% homogeneity. the purified enzyme shows multiple activities: 1) rnase activity on single-stranded, but not d ...19883286639
mitochondrial binding of a protein affected in mutants resistant to the microtubule inhibitor podophyllotoxin.specific antibodies to a protein p1 mr approximately equal to 63,000) from chinese hamster ovary cells, which is affected in mutants resistant to the microtubule inhibitor, podophyllotoxin, and behaves like a microtubule-related protein by certain criteria [14], have been raised. the antibody reacts specifically with the p1 protein in one- and two-dimensional immunoblots, and a cross-reacting protein of similar molecular mass and electrophoretic mobility is also found in cells from various verte ...19873319627
cloning, mapping and molecular analysis of the pyrg (orotidine-5'-phosphate decarboxylase) gene of aspergillus nidulans.we have modified the transformation procedures of ballance et al. [biochem. biophys. res. commun. 112 (1983) 284-289] to give increased rates of transformation in aspergillus nidulans. with the modified procedures we have been able to complement pyrg89, a mutation in the orotidine-5'-phosphate decarboxylase gene of a. nidulans, by transformation with a library of wild-type (wt) sequences in pbr329. we have recovered, by marker rescue from one such transformant, a plasmid (pjr15) that carries an ...19873328733
behaviour of recombinant plasmids in aspergillus nidulans: structure and stability.a pyrg- aspergillus strain was transformed with plasmid pdjb-1, derived from pbr325 by insertion of the neurospora crassa pyr4 gene (orotidine 5'-phosphate carboxylase), giving mitotically unstable transformants. aspergillus dna which acted as an "autonomously replicating sequence" (ars) in yeast was inserted into pdjb-1 and the resulting construct, pdjb12.1, gave mitotically stable transformants when introduced into aspergillus. transformants obtained with pdjb-1 and pdjb12.1 gave few pyr- prog ...19863329034
sequence organization and putative regulatory elements in the 5s rrna genes of two higher plants (vigna radiata and matthiola incana).the tandemly arranged and clustered highly repeated 5s rrna genes are investigated for two plants belonging to different higher plant families: matthiola incana (brassicaceae, dilleniidae, rosidae; 3600 5s rrna genes/n) shows a homogeneous repeat size of 510 bp, whereas vigna radiata (mung bean, former phaseolus aureus, fabaceae, rosidae; approx. 4300 5s rrna genes) has a repeat size of 215 bp. the mung-bean 5s rrna coding region starts 5' with agg and ends with cct; matthiola starts with ggg an ...19883371663
development of a homologous transformation system for aspergillus niger based on the pyrg gene.the development of a homologous transformation system for aspergillus niger is described. the system is based on the use of an orotidine-5'-phosphate decarboxylase deficient mutant (pyrg) and a vector, pab4-1, which contains the functional a. niger pyrg gene as a selection marker. transformation of the a. niger pyrg mutant with pab4-1 resulted in the appearance of stable pyr+ transformants at a frequency of 40 transformants per microgram of dna. in 90% of these transformants integration had occu ...19873472035
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