molecular cloning and analysis of abundant and stage-specific mrnas from puccinia characterize highly expressed mrnas from germinated urediniospores of puccinia graminis f. sp. tritici, we isolated 68 cdna clones of abundant mrna species belonging to at least six homology groups. the two most abundant homology groups, hg1 and hg2, contained 54 of the 68 cdna clones and accounted for 2.4 and 0.6% of the poly(a)+ rna in germinated urediniospores, respectively. by sampling different developmental stages of the uredinial cycle, we showed that the uam transcript, corresponding ...19938439672
the mitochondrial genome of the entomopathogenic fungus beauveria bassiana: analysis of the ribosomal rna region.the 28.5-kbp mitochondrial (mt) genome from the entomopathogenic fungus beauveria bassiana was studied using restriction enzyme analysis, gene probe hybridization, and dna sequence comparisons. a detailed restriction enzyme map allowed cloning of the entire genome into a number of segments. hybridization of heterologous gene probes to the mtdna resulted in the identification of the large ribosomal rna (lrrna) and small ribosomal rna (srrna) genes. gene probes derived from several yeasts and fung ...19938439871
cerato-ulmin, a toxin involved in dutch elm disease, is a fungal hydrophobin. 19938453298
aspergillus nidulans nuclear proteins bind to a ccaat element and the adjacent upstream sequence in the promoter region of the starch-inducible taka-amylase a gene.aspergillus nidulans was used as an intermediate host to investigate the regulation of the taka-amylase a (taa) gene from aspergillus oryzae. the induction of taa by starch was confirmed to be regulated at the transcriptional level by analyzing the transcripts specific for taa synthesized in vitro in nuclei from starch- and glucose-grown cells. a 55 bp dna fragment containing a consensus ccaat sequence from the promoter region of the taa gene was shown to confer starch inducibility on the gene. ...19938455560
evolutionary conservation of excision repair in schizosaccharomyces pombe: evidence for a family of sequences related to the saccharomyces cerevisiae rad2 gene.cells mutated at the rad13 locus in the fission yeast, schizosaccharomyces pombe are deficient in excision-repair of uv damage. we have cloned the s.pombe rad13 gene by its ability to complement the uv sensitivity of a rad13 mutant. the gene is not essential for cell proliferation. sequence analysis of the cloned gene revealed an open reading-frame of 1113 amino acids with structural homology to the rad2 gene of the distantly related saccharomyces cerevisiae. the sequence similarity is confined ...19938464724
carnitine acetyltransferase is absent from acuj mutants of aspergillus nidulans.the acuj mutant of aspergillus nidulans has been shown to lack carnitine acetyltransferase (cat) activity when grown under conditions where this activity is readily detectable in wild-type strains. revertants selected for growth on acetate recover cat activity and the ability to grow on long-chain fatty acids. when growing on carbon sources such as sucrose, cytosolic acetyl coenzyme a was generated by adenosine triphosphate (atp):citrate lyase.19938472927
properties and regulation of the cell cycle-specific nima protein kinase of aspergillus nidulans.nima is the protein product of the nima gene of the filamentous fungus aspergillus nidulans, required for progression of cells from g2 into mitosis. the protein kinase activity of nima, assayed by phosphorylation of beta-casein, varies during the nuclear division cycle, reaching a maximum in late g2 and m. to investigate the biochemical properties of this cell cycle-regulated protein kinase, we have expressed nima cdna that encodes full-length nima in escherichia coli as a fusion product with gl ...19938473320
properties of genes involved in the control of isocitrate lyase production in aspergillus nidulans.the interaction between genes of aspergillus nidulans conferring constitutive synthesis of isocitrate lyase (iclc a and iclcb) and fluoroacetate resistance (facb) has been investigated. although facb mutants are unable to induce the glyoxylate cycle enzyme isocitrate lyase in response to acetate as sole carbon source, this phenotype was suppressed in recombinants of the type iclc;facb. the iclca and iclcb mutations do not alter significantly the activities of eight enzymes of intermediary metabo ...19938473860
at least four regulatory genes control sulphur metabolite repression in aspergillus nidulans.mutations in four genes: scona (formerly sua25meth, mapa25), sconb (formerly mapb1), sconc and scond, the last two identified in this work, relieve a group of sulphur amino acid biosynthetic enzymes from methionine-mediated sulphur metabolite repression. exogenous methionine has no effect on sulphate assimilation in the mutant strains, whereas in the wild type it causes almost complete elimination of sulphate incorporation. in both mutant and wild-type strains methionine is efficiently taken up ...19938479426
specific binding sites in the alcr and alca promoters of the ethanol regulon for the crea repressor mediating carbon catabolite repression in aspergillus nidulans.the crea repressor responsible for carbon catabolite repression in aspergillus nidulans represses the transcription of the ethanol regulon. the n-terminal part of the crea protein encompassing the two zinc fingers (c2h2 class family) and an alanine-rich region was expressed in escherichia coli as a fusion protein with glutathione-s-transferase. our results show that crea is a dna-binding protein able to bind to the promoters of both the specific trans-acting gene, alcr, and of the structural gen ...19938483416
essential roles for calcium and calmodulin in g2/m progression in aspergillus nidulans.nimt encodes a protein in aspergillus nidulans that is required for tyrosine dephosphorylation of p34cdc2 and has a strong homology to cdc25-type proteins. conditional mutation of nimt (nimt23 mutation) arrests cells in g2 at the restrictive temperature. after release of the temperature-sensitive nimt23 block, p34cdc2 undergoes tyrosine dephosphorylation and we showed that as cells entered mitosis, a rapid increase in calmodulin was observed. the increase in calmodulin and progression into mitos ...19938486741
the aspergillus nidulans ya gene is regulated by abaa.the developmentally regulated aspergillus nidulans ya gene encodes a p-diphenol oxidase that is needed for synthesis of green conidial pigment. we subjected the ya 5' flanking region to mutational analysis in a. nidulans and saccharomyces cerevisiae to identify dna sequence elements involved in its transcriptional control, and identified two functionally distinct elements. element i contained potential brla binding sites and was required for full level ya transcription, but not for developmental ...19938491194
the aspergillus nidulans brla regulatory locus consists of overlapping transcription units that are individually required for conidiophore development.the aspergillus nidulans brla locus controls conidiophore development in conjunction with the products of several other regulatory loci. in this paper, we show that the brla locus consists of overlapping transcription units, designated alpha and beta, with alpha transcription initiating within beta intronic sequences. the predicted brla polypeptides differ by 23 amino acid residues at their n-termini. targeted mutations specifically eliminating either the alpha or beta transcript led to developm ...19938508769
translational repression of brla expression prevents premature development in aspergillus.the aspergillus nidulans brla developmental regulatory locus consists of two overlapping transcription units, brla alpha and brla beta, which encode functionally related polypeptides. we used translational fusions between each of the predicted brla reading frames and the escherichia coli lacz gene to test the hypothesis that developmental regulation of brla alpha and brla beta expression occurs through different mechanisms. brla alpha is transcriptionally controlled and a large portion of brla a ...19938508770
estimation of mitotic stability in conidial fungi: a theoretical framework.mitotic stability refers to the probability that genetic elements are transmitted to both daughters during mitosis. this is of practical importance in molecular genetics because autonomous cloning vectors should be transmitted at high frequency during mitosis. in filamentous coencytic fungi it is difficult to quantify mitotic stability because a fluctuation test is not feasible. we show how to get around this problem by formulating a general model of the transmission of nuclear genetic elements ...19938514143
the nitrate reductase gene from a shoyu koji mold, aspergillus oryzae kbn616.a niad gene encoding nitrate reductase was isolated from aspergillus oryzae kbn616 and sequenced. the structural gene comprises 2973 bp and 868 amino acids, which showed a high degree of similarity to nitrate reductases from other filamentous fungi. the coding sequence is interrupted by six introns varying in size from 48 to 98 bp. the intron positions are all conserved among the niad genes from a. oryzae, aspergillus nidulans, and aspergillus niger. a homologous transformation system was develo ...19958520125
cre1, the carbon catabolite repressor protein from trichoderma order to investigate the mechanism of carbon catabolite repression in the industrially important fungus trichoderma reesei, degenerated pcr-primers were designed to amplify a 0.7-bp fragment of the cre1 gene, which was used to clone the entire gene. it encodes a 402-amino acid protein with a calculated m(r) of 43.6 kda. its aa-sequence shows 55.6% and 54.7% overall similarity to the corresponding genes of aspergillus nidulans and a. niger, respectively. similarity was restricted to the aa-reg ...19958521952
dictyostelium myosin i double mutants exhibit conditional defects in pinocytosis.the functional relationship between three dictyostelium myosin is, myoa, myob, and myoc, has been examined through the creation of double mutants. two double mutants, myoa-/b- and myob-/c-, exhibit similar conditional defects in fluid-phase pinocytosis. double mutants grown in suspension culture are significantly impaired in their ability to take in nutrients from the medium, whereas they are almost indistinguishable from wild-type and single mutant strains when grown on a surface. the double mu ...19958522584
role of tryptophans in substrate binding and catalysis by dna photolyase. 19958524158
sok2 may regulate cyclic amp-dependent protein kinase-stimulated growth and pseudohyphal development by repressing transcription.yeast cyclic amp (camp)-dependent protein kinase (pka) activity is essential for growth and cell cycle progression. dependence on pka function can be partially relieved by overexpression of a gene, sok2, whose product has significant homology with several fungal transcription factors (stua from aspergillus nidulans and phd1 from saccharomyces cerevisiae) that are associated with cellular differentiation and development. deletion of sok2 is not lethal but exacerbates the growth defect of strains ...19958524252
gaip, a protein that specifically interacts with the trimeric g protein g alpha i3, is a member of a protein family with a highly conserved core domain.using the yeast two-hybrid system we have identified a human protein, gaip (g alpha interacting protein), that specifically interacts with the heterotrimeric gtp-binding protein g alpha i3. interaction was verified by specific binding of in vitro-translated g alpha i3 with a gaip-glutathione s-transferase fusion protein. gaip is a small protein (217 amino acids, 24 kda) that contains two potential phosphorylation sites for protein kinase c and seven for casein kinase 2. gaip shows high homology ...19958524874
characterization of the "promoter region" of the enolase-encoding gene enol from the anaerobic fungus neocallimastix frontalis: sequence and promoter analysis.the sequence of the neocallimastix frontalis enolase gene promoter was determined up to 1800 nucleotides 5' to the major transcriptional start point. the base composition of the enolase upstream sequence revealed a very a + t-rich profile (13.5% g + c) leading to many putative hairpin structures. the functional organization of the n. frontalis enolase promoter was investigated by heterologous transient-expression assays. dna fragments obtained by the sequential removal of sequences upstream of t ...19958536317
recombinational stability of replicating plasmids in aspergillus nidulans during transformation, vegetative growth and sexual reproduction.plasmids containing the ama1 replicon are capable of autonomous maintenance in aspergillus nidulans. it has been reported previously that these plasmids can form concatenates by recombination in a transformed mycelium, and up to 10% of molecules are involved in such events. the present study demonstrates that plasmid recombination, although frequent during transformation, rarely occurs during vegetative growth. as a result, the structure and phenotypic stability of ama1 plasmids generally remain ...19958536318
quantification of dna damage and repair in amino acid auxotrophs and uv-sensitive mutants of aspergillus nidulans using an elisa used to investigate dna repair in mammalian cells has been adapted to investigate mutagen-induced dna damage and repair in protoplasts of aspergillus nidulans. the assay shows a reduced rate of repair of dna damage in methionine and arginine auxotrophs (methg and argb), which were shown previously to be hypersensitive to uv radiation and chemical mutagens. the assay also showed a considerably reduced ability to repair mutagen-induced damage in the uv-sensitive mutants uvsb and uvsh. the ...19958543032
analysis of the regulation of the aspergillus nidulans penicillin biosynthesis gene aat (pende), which encodes acyl coenzyme a:6-aminopenicillanic acid acyltransferase.the regulation of the aspergillus nidulans penicillin biosynthesis gene aat (pende), which encodes acyl coenzyme a:6-aminopenicillanic acid acyltransferase (aat), was analysed. major transcriptional start sites map within 100 nucleotides upstream from the aat initiation codon. to study the regulation of aat expression, various aat-lacz gene fusions were constructed, in which the aat promoter region was fused in frame with the escherichia coli lacz reporter gene. a. nidulans strains carrying reco ...19958544821
cell cycle. the nima kinase joins forces with cdc2.the nima and cdc2 protein kinases cooperate to regulate mitosis in aspergillus nidulans. nima-related pathways have now begun to emerge in higher eukaryotes.19958548283
toxicology of halogenated aliphatic hydrocarbons: structural and molecular determinants for the disturbance of chromosome segregation and the induction of lipid peroxidation.the induction of mitotic chromosome malsegregation, mitotic arrest and lethality by a set of 55 halogenated hydrocarbons was investigated. to this aim, genetic assays in the mould aspergillus nidulans, able to provide precise quantitative information on the end-points studied, were used throughout the work. the experimental data obtained were used to develop qsar models for the induction of aneuploidy, which pointed to a major role of electrophilicity as molecular determinant for the aneugenic p ...19958548852
transport of lactate and its regulation in saccharomyces cerevisiae mutants deficient in specific metabolic steps. 19948550003
genetic and molecular characterisation of purine permease genes of aspergillus nidulans reveals a novel family of transporters conserved in prokaryotes and eukaryotes.the ascomycete fungus aspergillus nidulans can utilize purines (adenine, guanine, hypoxanthine, xanthine, and uric acid) as sole nitrogen sources [1]. the expression of most structural genes involved in the pathway of purine uptake and catabolism is subject to uric acid induction, mediated by the product of the positive regulatory gene uay, and to nitrogen metabolite repression, mediated by the product of the general, positive-acting, gata-like transcription factor, encoded by the area gene [1].19948550005
structure-function analysis of the proline permease (prnb) of the filamentous fungus aspergillus nidulans. 19948550020
enzymatic synthesis of hydrophobic penicillins.our knowledge of the enzymes and genes involved in the biosynthesis of beta-lactam antibiotics has increased notably in the last decade. the purification to homogeneity of some of these proteins as well as their biochemical characterization has allowed some of them to be used for synthesizing many different penicillins and cephalosporin-like products in vitro. in this report we describe the most important advances in this field, placing special emphasis on the enzymatic synthesis of hydrophobic ...19958557558
mutational analysis of the c-terminal region of area, the transcription factor mediating nitrogen metabolite repression in aspergillus aspergillus nidulans the positive-acting, wide domain regulatory gene area mediates nitrogen metabolite repression. previous analysis demonstrated that the c-terminal 153 residues of the area product (area) are inessential for at least partial expression of most genes subject to regulation by area. paradoxically, arear2, a -1 frameshift replacing the wild-type 122 c-terminal residues with a mutant peptide of 117 amino acids, leads to general loss of function. to determine the basis for the ar ...19968569680
allocation of random amplified polymorphic dna markers and enzyme activities to aspergillus nidulans and aspergillus tetrazonus chromosomes.chromosome-substituted haploid segregants of an a. nidulans x a. tetrazonus somatic hybrid were used to allocate several random amplified polymorphic dna and isoenzyme markers to parental chromosomes. twenty-six amplified dna fragments, and nine isoenzyme activities, including lactate dehydrogenase, superoxide dismutase, and arylesterase isoenzymes were assigned to chromosomes. chromosomes-specific markers were found for each a. nidulans and a. tetrazonus chromosome. these markers could be used ...19958572683
biosynthetic pathways of glycerol accumulation under salt stress in aspergillus nidulans.a culture of aspergillus nidulans (fgsc 359) was gradually adapted for growth in media containing up to 2 m nacl or was exposed to a salt shock with 2 m nacl. the intracellular glycerol level increased by about 7.9-fold in salt-adapted and 2.4-fold in salt-shocked cultures when compared to the unadapted culture. the biosynthetic pathway involved in the accumulation of glycerol was investigated under long-term salt adaptation and short-term salt shock. glycerol-3-phosphate dehydrogenase (ec 1.1.1 ...19958574901
integrative and replicative transformation of penicillium canescens with a heterologous nitrate-reductase gene.a wild isolate of penicillium canescens was subjected to mutagenesis, and 150 chlorate-resistant mutants were isolated and classified in respect of their ability to utilize various nitrogen sources. strains supposedly deficient in nitrate reductase have been transformed with the nitrate-reductase gene from aspergillus niger. transformation probably occurred by non-homologous integration of the transforming vector into the chromosome. co-transformation with the ama 1 replicating element from a. n ...19958575022
ethanol utilization regulatory protein: profile alignments give no evidence of origin through aldehyde and alcohol dehydrogenase gene fusion.the suggestion that the ethanol regulatory protein from aspergillus has its evolutionary origin in a gene fusion between aldehyde and alcohol dehydrogenase genes (hawkins ar, lamb hk, radford a, moore jd, 1994, gene 146:145-158) has been tested by profile analysis with aldehyde and alcohol dehydrogenase family profiles. we show that the degree and kind of similarity observed between these profiles and the ethanol regulatory protein sequence is that expected from random sequences of the same comp ...19958580855
domain structure and function within the quta protein of aspergillus nidulans: implications for the control of transcription.quta is a positively acting regulatory protein that regulates the expression of the eight genes comprising the quinic acid utilization gene (qut) gene cluster in aspergillus nidulans. it has been proposed that the quta protein is composed of two domains that are related to the n-terminal two domains-dehydroquinate (dhq) synthase and 5-enolpyruvyl shikimate-3-phosphate (epsp) synthase-of the pentadomain arom protein. the arom protein is an enzyme catalysing five consecutive steps in the shikimate ...19968581174
the aspergillus nidulans bime (blocked-in-mitosis) gene encodes multiple cell cycle functions involved in mitotic checkpoint control and mitosis.the bime (blocked-in-mitosis) gene appears to function as a negative mitotic regulator because the recessive bime7 mutation can override certain interphase-arresting treatments and mutations, causing abnormal induction of mitosis. we have further investigated the role of bime in cell cycle checkpoint control by: (1) coordinately measuring mitotic induction and dna content of bime7 mutant cells; and (2) analyzing epistasis relationships between bime7 and 16 different nim mutations. a combination ...19958586660
pneumonia due to fonsecaea pedrosoi and cerebral abscesses due to emericella nidulans in a bone marrow transplant recipient. 19958589181
optimization of glucose oxidase production by aspergillus niger using genetic- and process-engineering techniques.wild-type aspergillus niger nrrl-3 was transformed with multiple copies of the glucose oxidase structural gene (god). the gene was placed under the control of the gpda promoter of a. nidulans. for more efficient secretion the alpha-amylase signal peptide from a. oryzae was inserted in front of god. compared to the wild type, the recombinant strain nrrl-3 (god3-18) produced up to four times more extracellular glucose oxidase under identical culture conditions. addition of yeast extract (2 gl-1) t ...19958590664
generation and characterization of nadh: ubiquinone oxidoreductase mutants in neurospora crassa. 19958592454
organisation of the mitochondrial genome of trichophyton rubrum. dna sequence analysis of the nd4 gene, the atpase subunit-6 gene, the ribosomal rna small-subunit gene, the nd6 gene, the coxiii gene, the atpase subunit-8 gene and six trna genes that correspond respectively to the tyrosine, lysine, glutamine, asparagine, isoleucine and tryptophan isoacceptors.we present the nucleotide sequence of a 5207-bp-long region of the mitochondrial genome of the dermatophyte trichophyton rubrum. this represents about 1/5th of the total genome and extends a previous study. from the 5' end of the present sequence, the order of genes is as follows: the end of the nd4 gene, the gene coding for subunit 6 of atpase, the gene coding for the small ribosomal rna (ssu rrna), the tyrosyl trna gene, the nd6 gene, the coxiii gene, the atpase 8 subunit gene and a cluster of ...19958593686
transformations of penicillium islandicum and penicillium frequentans that produce anthraquinone-related compounds.wild-type strains of penicillium islandicum and penicillium frequentans, which produce anthraquinone and related compounds, were transformed to benomyl and hygromycin b resistance. plasmids psv50 and pbt6, with benomyl-resistant beta-tublin genes, and plasmids pan7-1 and pdh25, with a bacterial hygromycin phosphotransferase gene under the control of aspergillus nidulans sequences, were used respectively. transformation frequencies with these plasmids were 10-20 transformants per micrograms of dn ...19958593690
biolistic transformation of the obligate plant pathogenic fungus, erysiphe graminis f.sp. hordei.particle gun acceleration appears to be a possible way to transform mycelium cells of obligate plant parasites growing on host surfaces. gus expression was obtained in e. graminis f.sp. hordei cells after bombardment with the gus gene under the control of the e. graminis f.sp. hordei β-tubulin promoter. three heterologous promoters, onefrom aspergillus nidulans and two from cochliobolus heterostrophus, gave very low or no expression of gus.19958595653
two family g xylanase genes from chaetomium gracile and their expression in aspergillus nidulans.with oligonucleotides based on the amino-terminal and internal amino-acid sequences of a xylanase, two xylanase genes, cgxa and cgxb, were isolated and sequenced from chaetomium gracile wild and mutant strains. each gene isolated from both strains was essentially the same as far as nucleotide sequences were compared. the mature cgxa and cgxb xylanases comprise 189 and 211 amino acids, respectively, and share 68.5% homology. the cgxa was found to be the major enzyme in the mutant strain. comparis ...19958595661
wild chromosomal variants in aspergillus nidulans.pulsed-field gel electrophoresis and a chromosome-specific cosmid dna library were used to determine the karyotypes of wild-type aspergillus nidulans isolates from around the world. overall, little structural variation was found, with a few major exceptions. one isolate possessed a non-essential b-chromosome of about 1.0 million base pairs (mb). another isolate had undergone a non-reciprocal translocation of about 1.6 mb of chromosome vi onto chromosome viii. other than these chromosomal differe ...19968595677
mutational analysis reveals dispensability of the n-terminal region of the aspergillus transcription factor mediating nitrogen metabolite repression.mutational analysis has enabled identification and localization of an upstream exon of the area gene of aspergillus nidulans mediating nitrogen metabolite repression. a mutation in the initiation codon and frameshift mutations, which revert by restoration of the reading frame, established the coding role of the exon and mutations affecting intron splicing in conjunction with dna sequencing of reverse transcriptase polymerase chain reaction (rt-pcr) products localized the coding region intron. th ...19958596437
cata, a new aspergillus nidulans gene encoding a developmentally regulated catalase.aspergillus nidulans asexual sporulation (conidiation) is a model system for studying gene regulation and development. the can5 cdna is one of several clones isolated based on transcript induction during conidiation. here we present the molecular characterization of its corresponding gene, demonstrating that it encodes a developmentally regulated catalase, designated cata. the cata 744-amino-acid-residue polypeptide shows significant identity to other catalases. its similarity to prokaryotic cat ...19968598056
characterization of the ugata gene of ustilago maydis, isolated by homology to the gata gene of aspergillus nidulans.a gene encoding a putative gaba aminotransferase (ugata) was isolated from the basidiomycete ustilago maydis via heterologous hybridization to the gaba aminotransferase gene (gata) of aspergillus nidulans . the derived amino-acid sequence of ugata shows strong identity throughout the protein to the gaba aminotransferase enzymes from a. nidulans and saccharomyces cerevisiae. northern analysis in u. maydis indicated that the ugata transcript is inducible by the omega-amino acids gaba and beta-alan ...19968598057
regulation of beta-glucosidase biosynthesis in aspergillus nidulans.beta-glucosidase in aspergillus nidulans was found to be both intracellular and extracellular. the intracellular beta-glucosidase was synthesized after the exhaustion of carbon source in the medium. the extracellular enzyme appeared with autolysis of the mycelium. biosynthesis of beta-glucosidase was not induced by various carbohydrates but repressed to varying extents in the presence of glucose, glycerol, and 2-deoxyglucose. this repression was not relieved by addition of camp. the repression w ...19968598280
an alpha tubulin mutation suppresses nuclear migration mutations in aspergillus nidulans.microtubules and cytoplasmic dynein, a microtubule-dependent motor, are required for nuclei to move along the hyphae of filamentous fungi. nuclear migration in aspergillus nidulans is blocked by heat-sensitive (hs-) mutations in the nuda gene, which encodes dynein heavy chain, and the nudf gene, which encodes a g protein beta-subunit-like protein. hs- mutations in the nudc and nudg genes also prevent nuclear migration. we have isolated extragenic suppressor mutations that reverse the hs- phenoty ...19958601474
transformation of the cultivated mushroom, agaricus bisporus, to hygromycin b resistance.application of biotechnology to the cultivated mushroom, agaricus bisporus, has been hampered thus far by the lack of a transformation system. here, transformation of both a homo- and a heterokaryotic strain of a. bisporus to hygromycin b resistance is described. transforming dna was integrated into the a. bisporus genome and stably maintained throughout vegetative growth. transformants of the heterokaryotic strain formed transgenic fruiting bodies. promoters derived from the unrelated ascomycet ...19968602139
nucleotide sequence and expression of the gene encoding nadp+- dependent glutamate dehydrogenase (gdha) from agaricus bisporus.the gene encoding nadp+-dependent glutamate dehydrogenase (gdha) was isolated from an agaricus bisporus recombinant phage lambda library. the deduced amino acid sequence would specify a 457-amino acid protein that is highly homologous in sequence to those derived from previously isolated and characterized genes coding for microbial nadp+-gdh. the open reading frame is interrupted by six introns. none of the introns is located at either one of the positions of the two introns conserved in the cor ...19968602149
identification, cloning and analysis of the aspergillus niger gene pacc, a wide domain regulatory gene responsive to ambient ph.a wide domain regulatory gene implicated in modulating gene expression in response to ambient ph has been cloned and sequenced from the industrially useful filamentous fungus aspergillus niger. this gene, pacc, is able to restore a pacc+ phenotype to a. nidulans paccc11 and paccc14 mutants with respect to extent of conidiation, conidial pigment intensity and acid phosphatase regulation. the pacc gene of a. niger comprises three exons, encodes a three-zinc-finger protein of 677 amino acids, and s ...19968602152
gel mobility shift scanning of the acetate-inducible promoters from neurospora crassa reveals a common co-inducible dna-binding protein.the promoter regions of four acetate-inducible genes of neurospora crassa, acu-3, acu-5, acu-8 and acu-9, have been sequenced. using a scanning gel mobility shift assay particular dna regions in each promoter have been shown specifically to bind partially purified protein extracted from acetate-induced mycelia. the protein-binding regions so defined have common sequence motifs, elements of which are similar to those required for acetate induction in aspergillus nidulans.19968602159
a human peptidyl-prolyl isomerase essential for regulation of mitosis.the nima kinase is essential for progression through mitosis in aspergillus nidulans, and there is evidence for a similar pathway in other eukaryotic cells. here we describe the human protein pin1, a peptidyl-prolyl cis/trans isomerase (ppiase) that interacts with nima. ppiases are important in protein folding, assembly and/or transport, but none has so far been shown to be required for cell viability. pin1 is nuclear ppiase containing a ww protein interaction domain, and is structurally and fun ...19968606777
comparative analysis of the qutr transcription repressor protein and the three c-terminal domains of the pentafunctional arom enzyme.the arom protein is a pentadomain protein catalysing steps two to six in the prechorismate section of the shikimate pathway in microbial eukaryotes. on the basis of amino acid sequence alignments and the properties of mutants unable to utilize quinic acid as a carbon source, the arom protein has been proposed to be homologous throughout its length with the proteins regulating transcription of the genes necessary for quinate catabolism. the qutr transcription repressor protein has been proposed t ...19968611179
flug and flba function interdependently to initiate conidiophore development in aspergillus nidulans through brla beta activation.the aspergillus nidulans flug gene is necessary for the synthesis of a small diffusible factor that is required for the endogenously regulated induction of asexual sporulation that takes place during the development of an air-exposed colony. previous work established that flug is present at nearly constant levels throughout the aspergillus life cycle, leading to the hypothesis that flug factor is constitutively produced and development initiates after its concentration surpasses a fixed threshol ...19968617205
the 3-phosphoglycerate kinase gene of the yeast yarrowia lipolytica de-represses on gluconeogenic substrates.we have isolated the 3-phosphoglycerate kinase (pgk) gene of the yeast yarrowia lipolytica by probing a genomic library with a pcr fragment amplified with primers deduced from two highly conserved regions of various pgks. it is a unique sequence encoding a polypeptide of 417 residues with extensive homology to other pgks, especially to that of aspergillus nidulans (76% identity). the expression of the y. lipolytica pgk1 gene proved to be higher on gluconeogenic substrates than on glycolytic ones ...19968625424
evidence for slta1 as a salt-sensitive allele of the arginase gene (agaa) in the ascomycete aspergillus nidulans.strains of aspergillus nidulans carrying the slta1 mutation, conferring sensitivity to kcl and nacl, also showed an arginine-sensitive phenotype whereby concentrations of the l-amino acid at or above 10 mm were toxic to growth. sexual progeny of a cross between a slta1 mutant and a wild-type strain showed a co-segregation of salt and arginine sensitivity. similarly, revertants to salt tolerance showed a loss of arginine sensitivity as did slta1 strains that were transformed with a cosmid carryin ...19968625426
identification and cloning of a mobile transposon from aspergillus niger var. awamori.aspergillus niger var. awamori contains multiple copies of a transposable element, vader. this element was detected as a 437-bp insertion in four independently isolated spontaneous mutants of the niad (nitrate reductase) gene. the vader element is present in approximately 15 copies in both a. niger var. awamori and a. niger. a single copy of vader was detected from only one of the two laboratory strains of a. nidulans which were also examined. insertion of the vader element into the niad gene of ...19968625427
expression of an erwinia pectate lyase in three species of aspergillus.transgenic filamentous fungi of the species aspergillus niger, a. nidulans and a. awamori expressing and secreting erwinia carotovora subsp. atroseptica pectate lyase 3 (pl3) were generated. correct processing of the pre-enzyme was achieved using the a. niger pectin lyase a (pel a) signal peptide. with the prepro-peptide of a. niger polygalacturonase ii, secreted enzymes still possessed the 6- aa pro-sequence, indicating the importance of the conformation of the precursor protein for correct cle ...19968625428
autonomously replicating plasmids carrying the ama1 region in penicillium chrysogenum.plasmid vectors containing the ama1 sequence transformed with high efficiency and replicated autonomously in penicillium chrysogenum. the efficiency of transformation of p. chrysogenum was related to the length of the ama1 fragment used for constructing the different autonomously replicating plasmids. one of the two palindromic inverted repeats of ama1 (the 2.2-kb sali-hindiii fragment) is sufficient to confer autonomous replication and a high transformation efficiency. deletion of the 0.6-kb ce ...19968625429
endoglucanase ii (egii) of penicillium janthinellum: cdna sequence, heterologous expression and promotor analysis.the cdna coding for the endoglucanase egii of p. janthinellum was cloned and sequenced. the open reading frame comprises 1230 nucleotides and the deduced amino-acid sequence shows an overall homology of 63% with the t. reesei egl2. the cellulose-binding domain of egii represents a typical member of the a family of cellulases. the egl2 gene is only induced by cellulose or cellobiose and not by sophorose. a promotor fragment including 1 kb was cloned and sequenced. three major transcription startp ...19968625430
mutations affecting extracellular protease production in the filamentous fungus aspergillus nidulans.the extracellular proteases of aspergillus nidulans are known to be regulated by carbon, nitrogen and sulphur metabolite repression. in this study, a mutant with reduced levels of extracellular protease was isolated by screening for loss of halo production on milk plates. genetic analysis of the mutant showed that it contains a single, recessive mutation, in a gene which we have designated xpre, located on chromosome vi. the xpre1 mutation affected the production of extracellular proteases in re ...19968628232
purification and characterization of mitochondrial ribonuclease p from aspergillus nidulans.mitochondrial ribonuclease (rnase) p from aspergillus nidulans was purified to near homogeneity using whole-cell extract as the starting material. a 4400-fold purification with a yield of 5.2% was achieved by ammonium sulfate fractionation, heat treatment, and five types of column chromatography, including trna-affinity column chromatography. this enzyme, which has a molecular mass of 232 kda determined by glycerol gradient sedimentation analysis, appears to be composed of seven polypeptides and ...19968631344
the rna component of mitochondrial ribonuclease p from aspergillus nidulans.several rna molecules that copurified with aspergillus nidulans mitochondrial ribonuclease (rnase) p were identified [lee, y c., lee, b. j., hwang, d. s. & kang, h. s. (1996) eur j. biochem. 235, 289-296], and their partial sequences were determined. using an oligonucleotide probe, we cloned and mapped the gene encoding this putative rna component of rnase p (rnase p-rna), situated between urfa3 (unidentified reading frame a3) and coba (apocytochrome b) genes in the mitochondrial genome of a. ni ...19968631345
twenty-five coregulated transcripts define a sterigmatocystin gene cluster in aspergillus nidulans.sterigmatocystin (st) and the aflatoxins (afs), related fungal secondary metabolites, are among the most toxic, mutagenic, and carcinogenic natural products known. the st biosynthetic pathway in aspergillus nidulans is estimated to involve at least 15 enzymatic activities, while certain aspergillus parasiticus, aspergillus flavus, and aspergillus nomius strains contain additional activities that convert st to af. we have characterized a 60-kb region in the a. nidulans genome and find it contains ...19968643646
expression of a large number of novel testis-specific genes during spermatogenesis coincides with the functional reorganization of the male germ cell.structural and functional changes, essential for the formation of mature male germ cells, are known to take place at specific stages of the mammalian spermatogenic process. to identify novel genes that are involved in this developmental process, we have initiated a large-scale cdna sequencing project (hoog+, nucleic acids res. 19: 93-98, 1991; starborg et al., mol. reprod. dev. 33: 243-251, 1992; yuan et al., biol. reprod., 1995). five-hundred and forty cdnas have been isolated from testicular c ...19958645556
regulation of folate-dependent enzyme levels in aspergillus nidulans: studies with regulatory mutants.the synthesis of folate-dependent enzymes in aspergillus nidulans appears to be regulated by intracellular pools of homocysteine and methionine. the results are consistent with the view that homocysteine acts as an inducer and methionine as a corepressor, but the molecular mechanism of the regulation is still unknown. methionine-requiring mutants, meth2 and metd10, apparently allelic, show deregulation of folate-dependent enzymes. most characteristic of the mutants is a repressed level of folylp ...19968645712
extragenic suppressors of nudc3, a mutation that blocks nuclear migration in aspergillus nidulans.nuclear migration plays an important role in the growth and development of many organisms including the filamentous fungus aspergillus nidulans. we have cloned three genes from a. nidulans, nuda, nudc, and nudf, in which mutations affect nuclear migration. the nuda gene encodes the heavy chain of cytoplasmic dynein. the nudc gene encodes a 22-kd protein. the nudf gene was identified as an extra copy suppressor of the temperature sensitive (ts-) nudc3 mutation. the nudc3 mutation substantially de ...19958647384
nitrogen metabolite signalling involves the c-terminus and the gata domain of the aspergillus transcription factor area and the 3' untranslated region of its mrna.area is a gata transcription factor which mediates nitrogen metabolite repression in aspergillus nidulans in response to intracellular glutamine levels. we have identified and localized three elements important to modulation of area function: a region of 13 residues within the dna-binding gata domain which forms a putative extended loop structure, the 12 c-terminal residues, and sequences within a 218 nucleotide region of the 3' utr. the 12 c-terminal residues are also required for transcription ...19968654376
tyrosine 495 is a key residue in the active site of galactose oxidase. 19958654695
the tama gene of aspergillus nidulans contains a putative zinc cluster motif which is not required for gene function.expression of many nitrogen catabolic enzymes is controlled by nitrogen metabolite repression in aspergillus nidulans. although the phenotypes of tama mutants have implicated this gene in nitrogen regulation, its function is unknown. we have cloned the tama gene by complementation of a new tama allele. the tama sequence shares significant homology with the uga35/dal81/durl gene of saccharomyces cerevisiae. in vitro mutagenesis of sequences encoding a putative zinc cluster dna binding domain indi ...19968655534
[test for the effects of mutagenic toxic fungal metabolites originating from municipal landfill sites].the purpose of the study is to assess the mutagenic effect of mycotoxins produced by moulds growing on municipal landfill sites. mutagenicity of toxic fungal metabolites was determined by the salmonella plate incorporation assay with two strains of bacteria: ta98 and ta100, with and without metabolic activation. the results obtained indicate that there is a severe hazard caused by these mycotoxins detected main by ta98 with metabolic activation. the most mutagenic mixture of mycotoxins acting di ...19968656997
identification of a new antifungal target site through a dual biochemical and molecular-genetics approach.the target site of the antifungal compound ly214352 [8-chloro-4-(2-chloro-4-fluorophenoxy) quinoline] has been identified through a dual biochemical and molecular-genetics approach. in the molecular-genetics approach, a cosmid library was prepared from an aspergillus nidulans mutant that was resistant to ly214352 because of a dominant mutation in a single gene. a single cosmid (6a6-6) that could transform an ly214352-sensitive strain of a. nidulans to ly214352-resistance was isolated from the li ...19968660469
phylogenetic analysis of the isopenicillin-n-synthetase horizontal gene transfer.a phylogenetic study of the isopenicillin-n-synthetase (ipns) gene sequence from prokaryotic and lower eukaryotic producers of beta-lactam antibiotics by means of a maximum-likelihood approach has been carried out. after performing an extensive search, rather than invoking a global molecular clock, the results obtained are best explained by a model with three rates of evolution. grouped in decreasing order, these correspond to a. nidulans and then to the rest of the eukaryotes and prokaryotes, r ...19968662005
conservation of structure and function of the aflatoxin regulatory gene aflr from aspergillus nidulans and a. flavus.under limiting growth conditions, aspergillus nidulans produces a carcinogenic secondary metabolite related to aflatoxin and called sterigmatocystin (st). the genes for st biosynthesis are co-ordinately regulated and are all found within an approximately 60-kilobase segment of dna. one of the genes within this region is predicted to encode a cx2cx6cx6cx2cx6cx2 zinc binuclear cluster dna-binding protein that is related to the aspergillus flavus and aspergillus parasiticus aflatoxin regulatory gen ...19968662194
transformation of the plant pathogenic fungus, rhynchosporium secalis.the barley leaf scald fungus, rhynchosporium secalis, was transformed to hygromycin-b and phleomycin resistance using the hph gene from e. coli and the ble gene from streptoalloteichus hindustanus under the control of aspergillus nidulans promoter and terminator sequences. plasmid dna was introduced into fungal protoplasts by peg/cacl2 treatment. transformation frequencies varied from 59 to 493 transformants per 10 microg of dna and 5 x 10(7) protoplasts. the antibiotic-resistant phenotype appea ...19968662199
characterization of the aspergillus parasiticus niad and niia gene cluster.the nitrate reductase gene (niad) and nitrite reductase gene (niia) of aspergillus parasiticus are clustered and are divergently transcribed from a 1.6-kb intergenic region (niad-niia). the deduced aminoacid sequence of the a. parasiticus nitrate reductase demonstrated a high degree of homology to those of other aspergillus species, as well as to leptosphaeria maculans, fusarium oxysporum, gibberella fujikuroi and neurospora crassa, particularly in the cofactor-binding domains for molybdenum, he ...19968662212
in vitro activity of 1,3-beta-d-glucan synthase requires the gtp-binding protein the yeast saccharomyces cerevisiae, the family of rho genes are implicated in the control of morphogenetic events although the molecular targets of these gtp-binding proteins remain largely unknown. the activity of 1,3-beta-d-glucan synthase, the product of which is essential for cell wall integrity, is regulated by a gtp-binding protein, which we here present evidence to be rho1p. rho1p was found to copurify with fks1p, a glucan synthase subunit, in preparations of the enzyme purified by pro ...19968662910
purification and characterization of beta 2-tomatinase, an enzyme involved in the degradation of alpha-tomatine and isolation of the gene encoding beta 2-tomatinase from septoria lycopersici.lycopersicon species often contain the toxic glycoalkaloid alpha-tomatine, which is proposed to protect these plants from general microbial infection. however, fungal pathogens of tomato often are tolerant to alpha-tomatine and detoxification of alpha-tomatine may be how these pathogens avoid this potential barrier. as an initial step to evaluate this possibility, we have purfied to homogeneity a beta-1,2-d glucosidase from the tomato pathogen septoria lycopersici that hydrolyzes the beta-1,2-d ...19958664504
control of metabolic flux through the quinate pathway in aspergillus nidulans.the quinic acid ulitization (qut) pathway in aspergillus nidulans is a dispensable carbon utilization pathway that catabolizes quinate to protocatechuate via dehydroquinate and dehydroshikimate(dhs). at the usual in vitro growth ph of 6.5, quinate enters the mycelium by means of a specific permease and is converted into pca by the sequential action of the enzymes quinate dehydrogenase, 3-dehydroquinase and dhs dehydratase. the extent of control on metabolic flux exerted by the permease and the t ...19968670107
two s-phase checkpoint systems, one involving the function of both bime and tyr15 phosphorylation of p34cdc2, inhibit nima and prevent premature mitosis.we demonstrate that there are at least two s-phase checkpoint mechanisms controlling mitosis in aspergillus. the first responds to the rate of dna replication and inhibits mitosis via tyrosine phosphorylation of p34cdc2. cells unable to tyrosine phosphorylate p34cdc2 are therefore viable but are unable to tolerate low levels of hydroxyurea and prematurely enter lethal mitosis when s-phase is slowed. however, if the nima mitosis-promoting kinase is inactivated then non-tyrosine-phosphorylated p34 ...19968670863
quantitative analysis of gene expression in sexual structures of aspergillus nidulans by sequencing of 3'-directed cdna clones.we constructed a 3'-directed cdna library of cleistothecia and hülle cells of aspergillus nidulans to examine gene expression patterns of the sexual structures and to have probes necessary to isolate sexual structure-specific genes. sequencing of 360 randomly selected cdna clones yielded 272 expressed sequence tags (ests), most of which probably represent frequently or less expressed genes in sexual structures of a. nidulans. among the 272 ests, 33 ests (87 cdna clones) appeared more than once a ...19968674973
characterisation of the aspergillus nidulans fra1 mutant: hexose phosphorylation and apparent lack of involvement of hexokinase in glucose repression.hexose phosphorylation was studied in aspergillus nidulans wild-type and in a fructose non-utilising mutant (fra). the data indicate the presence of at least one hexokinase and one glucokinase in wild-type a. nidulans, while the fra1 mutant lacks hexokinase activity. the a. nidulans gene encoding hexokinase was isolated by complementation of the fra1 mutation. the absence of hexokinase activity in the fra1 mutant did not interfere with glucose repression of the enzymes involved in alcohol and l- ...19968674991
identification of a major cis-acting dna element controlling the bidirectionally transcribed penicillin biosynthesis genes acva (pcbab) and ipna (pcbc) of aspergillus nidulans.the beta-lactam antibiotic penicillin is produced as a secondary metabolite by some filamentous fungi. in this study, the molecular regulation of the aspergillus (emericella) nidulans penicillin biosynthesis genes acva (pcbab) and ipna (pcbc) was analyzed. acva and ipna are divergently oriented and separated by an intergenic region of 872 bp. translational fusions of acva and ipna with the two escherichia coli reporter genes lacz and uida enabled us to measure the regulation of both genes simult ...19968682797
beta-lactams. 19958688626
molecular characterisation of meab, a novel gene affecting nitrogen metabolite repression in aspergillus nidulans.mutations within the meab gene elicit the inappropriate expression of several activities subject to nitrogen metabolite repression in aspergillus nidulans and also have some unrelated phenotypic effects. we have cloned meab and isolated a full length cdna clone. northern analysis has shown that meab expression is not subject to nitrogen metabolite repression. meab encodes a novel protein of 418 amino acids and contains a significantly high number of s/tpxx motifs, a motif common in transcription ...19968690087
the detection and evaluation of aneugenic chemicals.although aneuploidy makes a significant contribution to both somatic and inherited disease the mechanisms by which environmental chemicals may induce numerical chromosome aberrations are only poorly defined. the european union project was aimed to further our understanding of those chemical interactions with the components of the mitotic and meiotic cell division cycle which may lead to aneuploidy and to characterise the parameters such as cellular metabolism which may influence the activity of ...19968692188
the genes yni1 and ynr1, encoding nitrite reductase and nitrate reductase respectively in the yeast hansenula polymorpha, are clustered and co-ordinately regulated.the nitrite reductase-encoding gene (yni1) from the yeast hansenula polymorpha was isolated from a lambda embl3 h. polymorpha genomic dna library, using as a probe a 481 bp dna fragment from the gene of aspergillus nidulans encoding nitrite reductase (niia). an open reading frame of 3132 bp, encoding a putative protein of 1044 amino acids with high similarity with nitrite reductases from fungi, was located by dna sequencing in the phages lambdanb5 and lambdaja13. genes yni1 and ynr1 (encoding ni ...19968694791
transformation of aspergillus nidulans by rna from rat macrophages stimulated with lipopolysaccharide.exogenous rna molecules can be incorporated into eukaryotic cells and can exert a variety of biological effects. we have previously described a model system for correcting genetic alterations of an aspergillus nidulans mutant using homologous rna and this phenomenon was named retrotransformation. in the present study, the retrotransformation of a. nidulans was performed with heterologous rna which was extracted from rat macrophages stimulated with lipopolysaccharide (lps). protoplasts of a. nidu ...19968696260
a novel family of developmentally regulated mammalian transcription factors containing the tea/atts dna binding domain.we describe the molecular cloning of two novel human and murine transcription factors containing the tea/atts dna binding domain and related to transcriptional enhancer factor-1 (tef-1). these factors bind to the consensus tea/atts cognate binding site exemplified by the gt-iic and sph enhansons of the sv40 enhancer but differ in their ability to bind cooperatively to tandemly repeated sites. the human tefs are differentially expressed in cultured cell lines and the mouse (m)tefs are differentia ...19968702974
the quta activator and qutr repressor proteins of aspergillus nidulans interact to regulate transcription of the quinate utilization pathway genese.genetic evidence suggests that the activity of the native quta transcription activator protein is negated by the action of the qutr transcription repressor protein. when aspergillus nidulans was transformed with plasmids containing the wild-type quta gene, transformants that constitutively expressed the quinate pathway enzymes were isolated. the constitutive phenotype of these transformants was associated with an increased copy number of the transforming quta gene and elevated quta mrna levels. ...19968704987
the ribosome-inactivating protein restrictocin deters insect feeding on aspergillus restrictus.the fungus-feeding beetle, carpophilus freemani, consumed equal quantities of young mycelia, fewer phialides bearing mature spores and much fewer phialides bearing developing spores of aspergillus restrictus compared to those of aspergillus nidulans when tested in diet choice assays. the degree to which specific fungal structures were consumed was inversely related to the localization of high levels of restrictocin, a ribosome-inactivating protein, to those structures. pure restrictocin added to ...19968704996
the aspergillus nidulans penicillin-biosynthesis gene aat (pende) is controlled by a ccaat-containing dna element.analysis of the promoter of the penicillin biosynthesis aat (pende) gene of aspergillus nidulans using band-shift assays led to the identification of a ccaat-containing dna element which was specifically bound by a protein (complex). the identified dna element was localised about 250 bp upstream of the transcriptional-start sites of aat. substitution of the ccaat core sequence by gatcc led to a fourfold reduction of expression of an aat-lacz gene fusion. the identified binding site thus was func ...19968706667
the hapc gene of aspergillus nidulans is involved in the expression of ccaat-containing promoters.the 5' regulatory region of the amds gene of aspergillus nidulans, which encodes an acetamidase required for growth on acetamide as a carbon and nitrogen source, contains a ccaat sequence which is required for setting the basal level of amds expression. mobility shift studies have identified a factor in a. nidulans nuclear extracts which binds to this ccaat sequence. in saccharomyces cerevisiae the hap3 gene encodes one component of a multisubunit complex that binds ccaat sequences. a search of ...19968709944
the aspergillus nidulans genes chsa and chsd encode chitin synthases which have redundant functions in conidia formation.we previously isolated three chitin synthase genes (chsa, chsb, and chsc) from aspergillus nidulans. in the present work, we describe the isolation and characterization of another chitin synthase gene, named chsd, from a. nidulans. its deduced amino acid sequence shows 56.7% and 55.9% amino acid identity, respectively, with cal1 of saccharomyces cerevisiae and chs3 of candida albicans. disruption of chsd caused no defect in cell growth or morphology during the asexual cycle and caused no decreas ...19968709948
the glucose repressor gene cre1 of trichoderma: isolation and expression of a full-length and a truncated mutant form.the cre1 genes of the filamentous fungi trichoderma reesei and t. harzianum were isolated and characterized. the deduced crei proteins are 46% identical to the product of the glucose repressor gene crea of aspergillus nidulans, encoding a dna-binding protein with zinc fingers of the c2h2 type. the cre1 promoters contain several sequence elements that are identical to the previously identified binding sites for a. nidulans crea. steady-state mrna levels for cre1 of the t. reesei strain qm9414 var ...19968709949
regulation of acid phosphatases in an aspergillus niger pacc disruption aspergillus niger strain has been constructed in which the ph-dependent regulatory gene, pacc, was disrupted. the pacc gene of a. niger, like that of a. nidulans, is involved in the regulation of acid phosphatase expression. disruptants were identified by a reduction in acid phosphatase staining of colonies. southern analysis demonstrated integration of the disruption plasmid at the pacc locus and northern analysis showed that the disruption strain produced a truncated pacc mrna of 2.2 kb (as ...19968709960
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