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the effects of putrescine, spermidine, and spermine on the growth of a polyamine-requiring mutant of aspergillus nidulans. 1976798706
presence of histones in aspergillus nidulans.five major histone proteins have been extracted from chromatin isolated from purified nuclei of the fungus, aspergillus nidulans. these proteins had chromatographic properties which were similar to reference calf thymus histones and were purified to electrophoretic homegeneity by gel chromatography of bio-gel p10, bio-gel p60, and sephadex g-100. electrophoresis of these proteins in three different systems (urea-starch, urea-acetic acid polyacrylamide, and discontinuous sds polyacrylamide) showe ...1976799641
choline-o-sulphate utilization in aspergillus nidulans. 1976800000
maternal inheritance of extranuclear mitochondrial markers in aspergillus nidulans. 1976800001
the inheritance of penicillin titre in wild-type isolates of aspergillus nidulans.in a sample of 52 wild-type isolates of aspergillus nidulans, penicillin titre ranged from 0-0 to 14-4 units/ml. these differences in titre were under genetic control. most of the variation between isolates was attributable to differences between heterokaryon-compatibility groups although significant differences were also found within groups. genetic variation for penicillin titre was observed among the progeny in four of seven crosses between wild-type isolates. in these four crosses the variat ...1975803549
the design of fermentation and biological assay procedures for assessment of penicillin production in populations of aspergillus nidulans. 1975805120
the fine structure of the branched alpha-d-glucan from the blue-green alga anacystis nidulans: comparison with other bacterial glycogens and phytoglycogen.the fine structure of the glycogen from the blue-green alga anacystis nidulans has been examined. after selective hydrolysis of all (1 yields 6)-alpha-d linkages by a bacterial isoamylase, the resulting mixture of linear chains was subjected to gel permeation chromatography. for purposes of comparison, the glycogens from escherichia coli and arthrobacter sp., amylopectin, phytoglycogen from sweet corn, and shell-fish glycogen were treated similarly. the profiles of the unit chains of a nidulans ...1975806344
sequence studies on 16s ribosomal rna from a blue-green alga.the 16s ribosomal rna of the blue green alga anacystis nidulans has been characterized in terms of the oligomers generated by digestion with t1 ribonuclease. a. nidulans by this criterion is definitely a procaryote; being no more distant from bacilli or enterics than the latter two are from one another. a. nidulans appears to be somewhat more closely related to the bacilli than to the enterics.1975813006
hemolytic, cytotoxic and complement inactivating properties of extracts of different species of aspergillus.some of the biological properties of saline extracts of the mycelia of several species of the aspergillus genus, namely, a. fumigatus, a. flavus, a. niger, a. nidulans, a. parasitucus and a. glaucus, were studied. only the extract prepared from a. fumigatus was found to be hemolytic for sheep red blood cells. in contrast, all the extracts with the only exception of that of a. glaucus, had cytotoxic effects on vero cells. both, the hemolytic and cytotoxic constituents of the extracts were removab ...1975813147
counterimmunoelectrophoresis in the diagnosis of mycetoma and its sensitivity as compared to immunodiffusion.counterimmunoelectrophoresis used for the diagnosis of mycetoma was found superior to immunodiffusion (id) especially when using neat serum and 1:2 dilution (p=less than 0.0001 with neat serum; and less than 0.01 with 1:2 dilution). it is recommended for routine use in mycology laboratories for following up patients on treatment and for confirming the diagnosis of sera that are weakly positive by id.1975817400
[a genetic transfer system in a cyanophyte of the genus aphanocapsa].the possibility of a genetic transfer, through a process of transformation, has been shown in a unicellular, facultative chemoheterotroph blue-green algae, aphanocapsa 6714. similarly to the process described with anacystis nidulans, the recipient strain becomes competent when the growth reaches its second, slower, exponential phase. the genetic marker conferring resistance to p-f-phenylalanine has been transferred with a frequency 20 to 100 times higher than that of the spontaneous mutations. t ...1976817846
isolation and characterization of a cyanophage infecting the unicellular blue-green algae a. nidulans and s. cedrorum. 1976821247
functional 70s hybrid ribosomes from blue-green algae and bacteria.hybrid 70s ribosomes were produced by combing anacystis nidulans and escherichia coli 30s and 50s subunits. both the a. nidulans 30se. coli 50s and e. coli 30s- a. nidulans 50s hybrids were functional in synthesizing protein when tested in a standard in vitro amino acid incorporating system. both 70s hybrids were inhibited by streptomycin but the degree of inhibition was dependent upon the source of the 30s subunit. the ability to form functional 70s ribosomes from subunits of blue-green algae a ...1976822798
genetics of biosynthesis and overproduction of penicillin. 1976823642
[action of various beta(1-3)-d-glucanases on the wall of yeasts: taxonomic applications].purified beta-(1-3)-d-glucanases (from aspergillus nidulans, badidiomycetes sp. qm 806, trichoderma viride) are used to release protoplasts from various yeasts. two of them may prove taxonomic correlation. the enzyme of basidomycetes sp. releases protoplasts from ascomycetes, the enzyme of t. viride releases protoplasts from ascomycetes and heterobasidiomycetes; none of them acts on the basidiomycetes.1976826331
role of nicotinic acid in pyridoxine biosynthesis in aspergillus nidulans. 1976999690
a cis-dominant regulatory mutation affecting enzyme induction in the eukaryote aspergillus nidulans. 19751089206
increased fusion frequency of aspergillus nidulans protoplasts. 19751089544
glucose uptake by aspergillus nidulans, purification and properties of glucose binding protein. 19751089548
mutations which affect amino acid transport in aspergillus nidulans.mutants deficient in amino acid transport (aau), and unable to utilize l-glutamate as a sole carbon and nitrogen source, have been isolated. there are four unlinked genes involved: aaua, aaub, aauc and aaud. the transport levels of certain amino acids, and the growth characteristics on certain nitrogen and carbon sources and toxic amino acid analogues, indicate that: aauai has defective transport of acidic amino acids; aaubi has defective transport of acidic and neutral amino acids; aauci and aa ...19751089752
a gene cluster in aspergillus nidulans with an internally located cis-acting regulatory region.work reported here on the fungus aspergillus nidulans has provided the first definitive demonstration of operon-type organisation in an eukaryote genome. it has been shown that the prna and prnb genes concerned with proline metabolism form a gene cluster with the regulatory region lying between the two putative structural genes prna and prnb. regulatory mutations (prnd) probably leading to relief of carbon catabolite repression, map in between prna and prnb and are cis-dominant with respect to b ...19751089903
initiator constitutive mutation with an 'up-promoter' effect in aspergillus nidulans.a mutation leading to strong constitutivity for a uric acid-xanthine permease in the lower eukaryote aspergillus nidulans has been found to be tightly linked to the putative structural gene whose expression it controls in the cis configuration. in addition, it increases the maximal induced level of transport 2.5-fold. this is the first demonstration of an initiator or promoter mutation in an eukaryote.19751089904
autolytic enzymes in hyphae of aspergillus nidulans: their action on old and newly formed walls.walls, purified from hyphae of the ascomycete aspergillus nidulans, autolyzed on incubation and liberated glucose, mannose, galactose, n-acetylglucosamine, and soluble oligosaccharides. digestion proceeded at linear rates until approximately 3% of the wall polymers had been hydrolyzed and then slowed markedly. the change in the rate of autolysis was not due to loss of enzyme activity but was caused by the disappearance of a fraction of the wall which was highly susceptible to digestion. radioact ...19751090573
keratomycosis in lucknow. 19751092323
reciprocal translocations and translocation disomics of aspergillus and their use for genetic mapping.two new techniques are described for genetic mapping of reciprocal translocations in a. nidulans, which can be used to locate centromeres and meiotically unlinked markers. they both make use of unbalanced disomics from heterozygous translocation crosses. these are mainly hyperhaploids of two classes: either typical-looking n + 1 with a normal chromosome in addition to a haploid set containing the translocation, or translocation disomics. when large chromosome segments are involved, such disomics ...19751092596
effect of l-histidine on the catabolism of nitrogenous compounds in aspergillus nidulans. 19751094095
nitrate utilization and growth in biotin-deficient aspergillus nidulans.a marked increase in the cellular synthesis accompanied by a decrease in the total fatty acid content was observed when aspergillus nidulans was grown on nh4no3 as a sole nitrogen source, in the medium containing avidin. because of the increased uptake of nh4(plus), the level of nh4(plus), was lowered in the medium; as a result there is early uptake and assimilation of nitrate by a biotin-deficient culture as compared with the normal culture of a. nidulans. at about 17mm concentration, nh4(plus) ...19751097067
the effect of chloral hydrate upon mitosis in aspergillus nidulans. 19751097582
quantitative tissue invasion of the murine brain as a phenotypic marker of strain virulence in aspergillus nidulans.in this study the extent of fungal invasion in tissues of the target organs (brain, heart and kidney) has been quantitated histologically in dba/2j mice inoculated intravenously with 3 strains of aspergillus nidulans of different virulence. this was done to determine the relation between tissue invasion and strain virulence as determined by time-mortality bioassay. it was found that quantitatively tissue invasion by a. nidulans was target organ specific. relative tissue invasion within each of t ...19751099700
homolanthionine in fungi: accumulation in the methionine-requiring mutants of aspergillus nidulans.homolanthionine, a higher homologue of cystathionine, was found to accumulate in the mutants of aspergillus nidulan impaired in the synthesis of methionine from homocysteine. the additional introduction of mutation resulting in a block at cystathionine gamma-synthase but not at cystathionine beta-synthase abolishes accumulation of both homolanthionine and cystathionine. this suggests a possible participation of cystathionine gamma-synthase in homolanthionine synthesis.19751101592
mutants affecting histidine utilization in aspergillus nidulans. 19751102388
an easy way of obtaining aspergillus nidulans haploids in the parasexual cycle using n-glycosyl polifungin. 19751102390
enzymatic lesions in methionine mutants of aspergillus nidulans: role and regulation of an alternative pathway for cysteine and methionine synthesis.in aspergillus nidulans the pathway involving cystathionine formation is the main one for homocysteine synthesis. mutants lacking cystathionine gamma-synthase or beta-cystathionase are auxotrophs suppressible by: (i) mutations in the main pathway of cysteine synthesis (cysa1, cysb1, and cysc1), (ii) mutations causing stimulation of cysteine catabolism (su101), and (iii) mutations in a presumed regulatory gene (suameth). a relative shortage of cysteine in the first group of suppressors causes a d ...19751102536
mutations affecting mitotic recombination frequency in haploids and diploids of the filamentous fungus aspergillus nidulans.a haploid strain of asp. nidulans with a chromosome segment in duplicate (one in normal position on chromosome i, one translocated to chromosome ii) shows mitotic recombination, mostly by conversion, in ade in a frequency slightly higher than in the equivalent diploid. a method has been devised, using this duplication, for the selection of rec and uvs mutations. six rec mutations have been found which decrease recombination frequency in the haploid. one mutation selected as uv sensitive showed a ...19751102911
cytoplasmic and nuclear mutations to chloramphenicol resistance in aspergillus nidulans.two chloramphenicol resistance mutations out of 123 tested in aspergillus nidulans are inherited extranuclearly as judged by transmissibility in heterokaryons, lack of segregation at meiosis, and independent segregation from all of the eight nuclear linkage groups. they do not recombine with each other. however, experiments in collaboration with g. turner and r.t. rowlands show that they do recombine with cytoplasmic mutations to oligomycin resistance (rowlands and turner, 1973) and cold-sensiti ...19751102919
pyrimidine biosynthesis in aspergillus nidulans: isolation and preliminary characterisation of auxotrophic mutants.113 pyrimidine auxotrophs, unable to synthesise ump have been selected in aspergillus nidulans. these mutants can be classified by complementation into eight groups, and genetic analysis has shown that five loci are involved. one complex locus consists of the mutually complementing pyra, pyrb and pyrc groups, as well as the cis-dominant pyrn group, members of which do not complement with members of the a, b or c groups. pyra mutants have been shown to lack cpsase-ur, pyrb and pyrc mutants have b ...19751102932
proceedings: fungicides causing mitotic segregation in aspergillus diploids. 19751102965
amide utilization in aspergillus nidulans: evidence for a third amidase enzyme.a mutation in a gene designated gmda has been found to lead to loss of ability of aspergillus nidulans to use benzamide, phenylacetamide and several other amides as sole nitrogen sources for growth. the gmda1 lesion results in low levels of an enzyme, called the general amidase, which has acitivity for a wide range of amide substrates. this enzyme is reressed by certain nitrogen-containing metabolites, including ammonium, but is probably not regulated by induction or by carbon catabolite repress ...19751104771
studies of partially repressed mutants at the tama and area loci in aspergillus nidulans.mutants, designated tamar, have been isolated on the basis of simultaneous resistance to toxic analogues thiourea, aspartate hydroxamate and chlorate with l-alanine as the sole nitrogen source. tamar mutants are also resistant to methylammonium. this resistance of tamar mutants is correlated with partially repressed activity of a number of enzyme and transport systems regulated by ammonium. furthermore, tam-ar mutants have low nadp-glutamate dehydrogenase (nadp-gdh) activity and also efflux ammo ...19751105154
sexual differentiation in aspergillus nidulans: the requirement for manganese and its effect on alpha-1,3 glucan synthesis and degradation.aspergillus nidulans was grown on media with added amounts of manganese ranging from 0--2.5 mum. manganese deficiency prevented cleistothecium development, although good vegetative growth was retained. subsequent analysis of the mycelium produced under mn2+ deficient growth revealed that alpha-1,3 glucan, the man carbon and energy source for fructification, was virtually absent from the cell wall. several enzymes related to cell wall composition were investigated. beta-1,3 glucanase, and very re ...19751106342
formation of 2,4-dihydroxybenzoic acid & resorcinol as intermediates in the degradation of salicylic acid by aspergillus nidulans. 19751107218
the reconstitution of anacystis nidulans dna-dependent rna polymerase from its isolated subunits.the dna-dependent rna polymerase of the blud-green alga anacystis nidulans was reconstituted from its isolated subunits in the absence of urea. applying this technique the kinetics and the subunit requirements of the reconstitution process were analyzed. the results reveal differences with respect to the reconstitution of escherichia coli polymerase. reconstitution proceeds much more slowly in the case of the a. nidulans enzyme. reconstitution here is absolutely dependent on the presence of the ...1976814000
labeling the deoxyribonucleic acid of anacystis nidulans.analysis of cell-free extracts of anacystis nidulans disclosed the absence of both thymidine phosphorylase (ec 2.4.2.4) and thymidine kinase (ec 2.7.1.21) activities. thymine and thymidine were incorporated inefficiently by intact cells of a. nidulans either in the presence or absence of deoxyguanosine (250 mug/ml). deoxythymidine monophosphate incorporation was also inefficient. radioactive deoxyadenosine, at a minimally toxic level (3 mug/ml), was incorporated effectively into the deoxyribonuc ...1975809413
cytochrome abnormalities and cyanide-resistant respiration in extranuclear mutants of aspergillus nidulans.the cytochrome spectra of two extranuclear mutants of aspergillus nidulans and the double-mutant recombinant formed from them have been examined both at room temperature and at the temperature of liquid n2 and compared with those of the wild-type strain. the oligomycin-resistant, slow growing mutant contained an increased amount of cytochrome c without any loss of cytochromes b and a,a3. the cold-sensitive mutant, apparently normal when grown at 37 c, showed an increased amount of cytochrome c a ...19761107321
hybridization and selection for increased penicillin titre in wild-type isolates of aspergillus nidulans.repeated hybridization and selection among wild-type isolates produced strains of aspergillus nidulans with increased penicillin titre. four independent selection lines were established, each originating from a sexual cross between two different heterokaryon-incompatible wild-type isolates. in each generation, two selected high-titre sister strains were crossed to produce the next generation. an initial increase in titre was obtained in each line, but after four or five generations of selection ...19751107473
hybridization and selection for increased penicillin titre in wile-type isolates of aspergillus nidulans.repeated hybridization and selection among wild-type isolates produced strains of aspergillus nidulans with increased penicillin titre. four independent selection lines were established, each originating from a sexual cross between two different heterokaryon-incompatible wild-type isolates. in each generation, two selected high-titre sister strains were crossed to produce the next generation. an initial increase in titre was obtained in each line, but after four or five generations of selection ...19751107474
the inheritance of penicillin titre in crosses between lines of aspergillus nidulans selected for increased productivity.selection for increased penicillin production among the progeny of pairwise crosses between wild-type isolates of aspergillus nidulans resulted in the production of high-titre strains. three crosses were made between strains with increased titre derived from independent selection lines. significant genetic variation was found among the progeny of each cross, indicating that different genes for increased titre had been selected in each line. gene action was additive in each cross. renewed selecti ...19751107475
genetic control of chromosome instability in aspergillus nidulans as a mean for gene amplification in eukaryotic microorganisms.a haploid strain of aspergillus nidulans carrying i-ii duplication homozygous for the leaky mutation ade20 shows impreved growth on minimal medium. the duplication, though more stable than disomics, still shows instability. several methods were used for detecting genetic control of improved stability. (a) visual selection, using a duplicated strain which is very unstable due to uv sensitivity, (ade20, bial/dp ya2; uvsb). one stable strain showed a deletion (or a lethal mutation?) distal to bia o ...19751107799
cysteine biosynthesis in aspergillus nidulans.the existence of two postulated pathways of anabolic cysteine biosynthesis in aspergillus midulans was investigated. no activities of the postulated pathway involving s-sulfocysteine as intermediate have been detected. investigations on cyteine and methionine requiring mutants revealed independent regulation of o-acetylserine sulfhydrylase by endogeneous cysteine and methionine pools. the reaction catalysed by o-acetylserine sulfhydrylase is postulated as the only anabolic pathway of cysteine bi ...19751108603
an unstable strain of aspergillus foetidus segregating proline auxotrophs.two basic colony types have been obtained through single conidial isolation from the bode strain of aspergillus foetidus as well as from mutants of this unstable strain. type i is prototrophic whereas type ii is an auxotroph requiring proline. when a type i strain is grown on complex medium it gradually becomes overwhelmed by type ii sectors of growth. however, essentially pure cultures of type i can be maintained on minimal medium (lacking proline). the yield of glucoamylase from type ii cultur ...19751147611
mycotoxin-producing potential of mold flora of dried beans.to evaluate the potential for mycotoxin production by molds in dried beans, the mold flora of 114 samples was determined both before and after surface disinfection of the beans with 5% naocl. surface disinfection substantially reduced mold incidence, indicating that contamination was mainly on the surface. the flora, both before and after disinfection, was dominated by species of the aspergillus glaucus group, the toxicogenic species a ochracues, penicillium cyclopium, and p. viridicatum, and sp ...19751168442
tumor induction by a single subcutaneous injection of sterigmatocystin in newborn mice.sterigmatocystin, a mycotoxin produced by aspergillus versicolor, aspergillus sydowi, aspergillus nidulans, and a species of bipolaris, was given to newborn balb/c x dba/2f1 (hereafter referred to as cd2f1) mice by a single s.c. administration in 1% gelatin suspension. in an acute toxicity study, the maximum tolerated dose of sterigmatocystin was 5 mug/g body weight. in a chronic study, a single s.c. injection of 5, 1, or 0.5 mug/g body weight gave rise to high incidences of lung and liver adeno ...19761268822
tests for recombinagens in fungi.three types of mitotic recombination can be studied in aspergillus nidulans and saccharomyces cerevisiae: (1) the classical type of reciprocal mitotic crossing-over which can be detected when it occurs between non-sister chromatids at the four-strand stage followed by co-segregation of a crossing-over and a non-crossing-over chromatid in the subsequent mitotic division. consequently, mitotic crossing-over reflects cellular responses to primary genetic damage in the g2 phase of the cell cycle. (2 ...19921279386
immunogenic potential of aspergillus nidulans subcellular fractions and their polypeptide components.cell-free extracts of the ascomycetous fungus aspergillus nidulans were separated into three subcellular fractions: cell walls, total membranes and cytosol, and two different immunization protocols were used to raise antibodies against them in 12 new zealand rabbits. the immune response was followed over time by dot and western blot analyses to determine the immunogenic potential of each individual fraction and their polypeptide components. the igg fractions, purified from pools of the best sera ...19921291875
structure of the oxidized long-chain flavodoxin from anabaena 7120 at 2 a resolution.the structure of the long-chain flavodoxin from the photosynthetic cyanobacterium anabaena 7120 has been determined at 2 a resolution by the molecular replacement method using the atomic coordinates of the long-chain flavodoxin from anacystis nidulans. the structure of a third long-chain flavodoxin from chondrus crispus has recently been reported. crystals of oxidized a. 7120 flavodoxin belong to the monoclinic space group p2(1) with a = 48.0, b = 32.0, c = 51.6 a, and beta = 92 degrees, and one ...19921303762
roles of the orla, tse, and bimg genes of aspergillus nidulans in chitin synthesis.strains of aspergillus nidulans carrying the orla1 or tse6 allele are deficient in cell wall chitin and undergo lysis at restrictive temperatures. the strains are remediable by osmotic stabilizers or by the presence of n-acetylglucosamine (glcnac) in the medium. the remediation by glcnac suggests that the lesion(s) in chitin synthesis resides in the amino sugar biosynthetic pathway prior to the synthesis of n-acetylglucosamine-6-phosphate. orla1 strains grown at permissive temperature exhibit an ...19921309526
differential flux through the quinate and shikimate pathways. implications for the channelling hypothesis.the qutc gene encoding dehydroshikimate dehydratase has been constitutively overexpressed in aspergillus nidulans from a range of 1-30-fold over the normal wild-type level. this overexpression leads to impaired growth in minimal medium which can be alleviated by the addition of aromatic amino acids to the medium. overexpression of the qutc gene in mutant strains lacking protocatechuic acid (pca) oxygenase leads to the build up of pca in the medium, which can be measured by a simple assay. measur ...19921318019
analysis of promoter activity by transformation of acremonium chrysogenum.promoter activity was examined in the beta-lactam-producing fungus, acremonium chrysogenum, by assessment of the properties of transformant isolates. transformation was achieved using plasmid constructs specifying hygromycin b resistance (hyr) linked to the promoter elements of gpda (the glucose-6-phosphate dehydrogenase-encoding gene of aspergillus nidulans), and pcbc [the gene encoding the isopenicillin n synthetase (ipns) enzyme of a. chrysogenum]. transformation frequency, hyr levels, and hy ...19921318244
overproduction in escherichia coli of the dehydroquinate synthase domain of the aspergillus nidulans pentafunctional arom protein.the pentafunctional arom protein of aspergillus nidulans is encoded by the complex aroma locus and catalyses steps 2-6 in the synthesis of chorismate, the common precursor for the aromatic amino acids and p-aminobenzoic acid. dna sequences encoding the 3-dehydroquinate synthase (dhq synthase) and 3-dehydroquinase domains of the arom protein have been amplified with the inclusion of a translational stop codon at the c-terminus by pcr technology. these amplified fragments of dna have been subclone ...19921320381
purification and characterization of a mammalian endo-exonuclease.an endo-exonuclease has been purified from cultured monkey (cv-1) cells. the enzyme which was purified to near homogeneity to be a 65 kda monomeric protein. the single-strand dnase activity is endonucleolytic and nonprocessive, whereas the double-strand dnase activity is exonucleolytic and processive. the enzyme was also found to have rnase activity using poly-ra as substrate. the ph optimum for ss-dnase is 8 and for ds-dnase it is 7.5. both dnase activities require a divalent metal ion (mg2+, m ...19921324480
mitochondrial dna sequence analysis of the cytochrome oxidase subunit i and ii genes, the atpase9 gene, the nadh dehydrogenase nd4l and nd5 gene complex, and the glutaminyl, methionyl and arginyl trna genes from trichophyton rubrum.in this paper, we present the nucleotide sequence of a 5248 bp-long region of the mitochondrial (mt) genome of the dermatophyte trichophyton rubrum. this region which represents about 1/4 of the total mt genome of this species reveals a compact organization of genes including: the glutaminyl trna, the methionyl trna, the cytochrome oxidase subunit i gene, the arginyl trna, the mitochondrial version of the atpase subunit 9 gene, the cytochrome oxidase subunit ii gene and a part of the nadh dehydr ...19921326416
transformation of gibberella fujikuroi: effect of the aspergillus nidulans ama1 sequence on frequency and integration.a stable and reproducible transformation selection system for gibberella fujikuroi protoplasts based on the aspergillus nidulans arg b gene, encoding ornithine transcarbamylase, has been developed. inclusion into the vector of the a. nidulans dna fragment (ama1), which permits plasmid autonomous replication in a. nidulans, a. niger and a. oryzae, appeared to permit autonomous replication of g. fujikuroi although the transformation frequency was increased by only two-fold. transformation was also ...19921327547
inducible overproduction of the aspergillus nidulans pentafunctional arom protein and the type-i and -ii 3-dehydroquinases from salmonella typhi and mycobacterium tuberculosis.the aroq gene of mycobacterium tuberculosis, encoding a type-ii 3-dehydroquinase, and the arod gene of salmonella typhi, encoding a type-i 3-dehydroquinase, have been highly overexpressed in escherichia coli using the powerful trc promoter contained within the expression vector pkk233-2. the m. tuberculosis type-ii 3-dehydroquinase has been purified in bulk from overproducing strains of e. coli to greater than 95% homogeneity. the protein is extremely heat-stable, is active as a homododecamer an ...19921329726
cell division in aspergillus.amenable to sophisticated genetic and molecular analysis, the simple filamentous fungus aspergillus nidulans has provided some novel insights into the mechanisms and regulation of cell division. mutational analysis has identified over fifty genes necessary for nuclear division, nuclear movement and cytokinesis. molecular and cellular analysis of these mutants has led to the discovery of novel components of the cytoskeleton as well as to clarifying the role of established cytoskeletal proteins. m ...19921336015
structure of the cochliobolus heterostrophus glyceraldehyde-3-phosphate dehydrogenase gene.a single gene (gpd1) encoding glyceraldehyde-3-phosphate dehydrogenase (gpd) was found in cochliobolus heterostrophus. homology with other fungal gpd-encoding genes was substantial at both the nucleotide and amino-acid levels. positions of four introns found in gpd1 were conserved in the corresponding aspergillus nidulans gpda gene (which is known to have three additional introns absent in gpd1). the size (approximately 1300 nucleotides) of the single gpd1 transcript was consistent with the leng ...19921339326
a second gene (quth) within the aspergillus nidulans-quinic-acid utilisation gene cluster encodes a protein with a putative zinc-cluster motif.a sequence of 3299 nt, contiguous with the previously sequenced quinate permease-encoding (qutd) gene and encompassing the dehydroshikimate dehydratase-encoding (qutc) gene, has been determined. northern-blot analysis detected (i) a quinate-inducible mrna of the expected size for the qutc gene, and (ii) a quinate-inducible mrna of 1.45 kb divergently transcribed away from qutc towards qutd. computer-aided sequence analysis identified an orf of 1047 nt corresponding to the qutc gene encoding dehy ...19921339361
calmodulin and cell cycle control.previous studies have indicated a role for the calcium receptor calmodulin in the control of eukaryotic cell proliferation. using a molecular genetic approach in the filamentous fungus aspergillus nidulans we have shown that cam is required for cell cycle progression at multiple points in the cell cycle. construction of an a nidulans strain conditional for calmodulin expression reveals that this protein is required during g1/s and for the initiation of mitosis. a lack of calmodulin results in ce ...19921343599
genotoxicity of fungi evaluated by sos microplate assay.by an introduction of sodium dodecylsulfate for cell lysis and immunomicroplate for mass assay, the modified sos microplate assay method was established and applied for the evaluation of genotoxicity of mycotoxins and fungal cultures. among 20 mycotoxins, the carcinogenic dihydrobisfuranoids such as aflatoxin b1, sterigmatocystin, and versicolorin a were positive in the presence of the activation system. while, the carcinogenic anthraquinones and lactones such as luteoskyrin, rugulosin, ochratox ...19921344897
isolation and characterization of the acetyl-coa synthetase from penicillium chrysogenum. involvement of this enzyme in the biosynthesis of penicillins.acetyl-coa synthetase (acs) of penicillium chrysogenum was purified to homogeneity (745-fold) from fungal cultures grown in a chemically defined medium containing acetate as the main carbon source. the enzyme showed maximal rate of catalysis when incubated in 50 mm hcl-tris buffer, ph 8.0, at 37 degrees c. under these conditions, acs showed hyperbolic behavior against acetate, coa, and atp; the km values calculated for these substrates were 6.8, 0.18, and 17 mm, respectively. acs recognized as s ...19921347531
nucleotide sequence of a gene from phanerochaete chrysosporium that shows homology to the faca gene of aspergillus nidulans.heterologous hybridisation was used to isolate a genomic dna sequence from phanerochaete chrysosporium using the faca (acetyl coa synthetase) gene from aspergillus nidulans as a probe. the cloned sequence hybridises to a 2.2 kb transcript in poly(a)+ rna prepared from mycelium grown on acetate as the sole carbon source. comparison of the dna sequence obtained with those of the a. nidulans faca and n. crassa acu5 genes reveals an orf that appears to be interrupted by five typical fungal introns. ...19921352996
the acu-1 gene of coprinus cinereus is a regulatory gene required for induction of acetate utilisation enzymes.we have isolated a gene from coprinus cinereus which cross-hybridises to the faca and acu-5 genes of aspergillus nidulans and neurospora crassa, respectively. these genes encode acetyl-coa synthetase, an enzyme which is inducible by acetate and required for growth on acetate as sole carbon source. we have designated the c. cinereus gene acs-1 and have used transformation to demonstrate its functional homology to the ascomycete genes by complementation of an n. crassa acu-5 mutation. the acs-1 ge ...19921354839
mitochondrial dna of schizophyllum commune: restriction map, genetic map, and mode of inheritance.mitochondrial dna (mtdna) found in the basidiomycete schizophyllum commune (strain 4-40) is a circular molecule 49.75 kbp in length. a physical map containing 61 restriction sites revealed no repeat structures. cloned genes from neurospora crassa, aspergillus nidulans, and saccharomyces cerevisiae were used in southern hybridizations to locate nine mitochondrial genes, including a possible pseudogene of atpase 9, on the restriction map. a probe from a functional atpase 9 gene identified homologo ...19921358467
cloning and disruption of a gene required for growth on acetate but not on ethanol: the acetyl-coenzyme a synthetase gene of saccharomyces cerevisiae.a dna fragment of saccharomyces cerevisiae with high homology to the acetyl-coenzyme a (acetyl-coa) synthetase genes of aspergillus nidulans and neurospora crassa has been cloned, sequenced and mapped to chromosome i. it contains an open reading frame of 2139 nucleotides, encoding a predicted gene product of 79.2 kda. in contrast to its ascomycete homologs, there are no introns in the coding sequence. the first atg codon of the open reading frame is in an unusual context for a translational star ...19921363452
hen egg white lysozyme expressed in, and secreted from, aspergillus niger is correctly processed and folded.we transformed aspergillus niger with the full length cdna gene encoding hen egg-white lysozyme (hewl) and its secretion signal sequence. lysozyme levels up to 12 mg/l were secreted when expression was controlled by the a. awamori glucoamylase (gam) promoter and 1 mg/l when controlled by the a. nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter. n-terminal sequence analysis of the recombinant protein indicated that the signal peptide was correctly processed by the a. niger secretor ...19901366900
efficient kex2-like processing of a glucoamylase-interleukin-6 fusion protein by aspergillus nidulans and secretion of mature interleukin-6.we have designed an expression vector for the secretion of human interleukin-6 (hil-6) in which the mature protein is fused through a spacer peptide, containing a kex-2 like protein processing signal, to the entire aspergillus niger glucoamylase (glaa) gene. transformation of aspergillus nidulans with this vector results in fungal strains secreting equimolar amounts of the glucoamylase and il-6 proteins. the kex2-type processing signal, lys-arg, is recognized and cleaved efficiently by an enzyme ...19911367012
the ethanol utilization regulon of aspergillus nidulans: the alca-alcr system as a tool for the expression of recombinant proteins. 19911367013
expression of heterologous proteins in aspergillus.filamentous fungi, in particular those of the genus aspergillus have been well exploited for their ability to produce high levels of extracellular proteins in an inexpensive manner. since many human proteins with the potential to be used therapeutically are secreted and require post-translational modification for biological activity, eukaryotic expression-secretion systems have been targeted for development. recent developments in dna-mediated transformation systems have allowed the utilization ...19911367014
identification and expression of the acv synthetase gene.the gene coding for acv synthetase has recently been identified and cloned. analysis of its structure and expression, along with similar studies of other genes involved in beta-lactam biosynthesis, should lead to a better understanding of the molecular basis of regulation of the pathway and the possibility of modifying yield and diversity of fungal antibiotics.19911367017
construction and physiological characterization of glyceraldehyde-3-phosphate dehydrogenase overproducing transformants of aspergillus nidulans.the construction and characterization of glyceraldehyde-3-phosphate-dehydrogenase (gpd) overproducing transformants of aspergillus nidulans and their behaviour in acetate-limited continuous cultures and glucose-grown batch cultures are described. the a. nidulans acetamidase deletion strain mh1277 was transformed with the homologous gpda gene on a vector with the homologous acetamidase-gene (amds) as a selection marker. transformant a1 contains about nine integrated copies of the gpda gene, and s ...19911367201
integrative transformation of the ascomycete podospora anserina: identification of the mating-type locus on chromosome vii of electrophoretically separated chromosomes.protoplasts of wild-type strain s and a long-lived extrachromosomal mutant (al2) of the ascomycete podospora anserina were transformed using a plasmid (pan7-1) which contains the hygromycin b phosphotransferase gene (hph) of escherichia coli under the control of aspergillus nidulans regulatory sequences. after optimizing the transformation procedure, transformation efficiencies of 15-21 transformants/micrograms plasmid dna were obtained. using a second selectable vector (pbt3), which contains th ...19911367277
intracellular and extracellular production of proteins in aspergillus under the control of expression signals of the highly expressed aspergillus nidulans gpda gene.the expression in aspergillus is described of genes, coding for intracellular and extracellular proteins controlled by the promoter region of the constitutively and efficiently expressed glyceraldehyde-3-phosphate dehydrogenase gene (gpda) of aspergillus nidulans. both the homologous gpda and the heterologous escherichia coli beta-galactosidase (lacz) and beta-glucuronidase (uida) genes could be expressed intracellularly at levels as high as 10-25% of total soluble protein. efficient extracellul ...19911367494
molecular characterization and functional analysis in aspergillus nidulans of the 5'-region of the penicillium chrysogenum isopenicillin n synthetase gene.the isopenicillin n synthetase gene (pcbc) was isolated from a genomic library of penicillium chrysogenum bc39813, a penicillin production strain. the nucleotide sequence, including 555 bp upstream of the translation start site was determined. various deletions within the pcbc 5'-region were constructed and linked to the escherichia coli lacz gene. an aspergillus nidulans argb strain was transformed with dna of these constructions. the region essential for promoter function could be localized be ...19911367495
cloning of the nitrate-nitrite reductase gene cluster of penicillium chrysogenum and use of the niad gene as a homologous selection marker.a new homologous transformation system for the filamentous fungus penicillium chrysogenum is described. the system is based on complementation of niad mutants using the nitrate reductase structural gene (niad) of p. chrysogenum. spontaneous niad mutants were identified after selection for chlorate resistance, in growth tests and subsequent complementation with the niad gene of aspergillus oryzae. the p. chrysogenum niad gene was isolated from a genomic library using the aspergillus nidulans niad ...19911367546
clusters of genes for the biosynthesis of antibiotics: regulatory genes and overproduction of pharmaceuticals.in the last decade numerous genes involved in the biosynthesis of antibiotics, pigments, herbicides and other secondary metabolites have been cloned. the genes involved in the biosynthesis of penicillin, cephalosporin and cephamycins are organized in clusters as occurs also with the biosynthetic genes of other antibiotics and secondary metabolites (see review by martín and liras [65]). we have cloned genes involved in the biosynthesis of beta-lactam antibiotics from five different beta-lactam pr ...19921368054
mitotic gold in a mold: aspergillus genetics and the biology of mitosis.the analysis of fungal mutants has had an extraordinary impact on our understanding of the biochemistry and regulation of mitosis. in this article we review the contribution of work on the filamentous fungus aspergillus nidulans to the molecular genetics of mitosis.19921369734
l-lysine repression of penicillin biosynthesis and the expression of penicillin biosynthesis genes acva and ipna in aspergillus nidulans.the addition of 0.1 m l-lysine to the fermentation medium reduced the production of penicillin by about 50% in aspergillus nidulans. to analyse this effect at the molecular level, the expression of the penicillin biosynthesis genes acva and ipna, encoding delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine synthetase and isopenicillin n synthetase, was studied by using translational fusions with different reporter genes (strain axb4a, acva-uida, ipna-lacz fusions; axb4b, acva-lacz, ipna-uida fusion ...19921369977
the induction of mitotic chromosome malsegregation in aspergillus nidulans. quantitative structure activity relationship (osar) analysis with chlorinated aliphatic hydrocarbons.the biological activity of 24 chlorinated aliphatic hydrocarbons has been studied in the mold aspergillus nidulans. the ability to induce chromosome malsegregation, lethality and mitotic growth arrest has been experimentally determined for each chemical. these data, together with those of 11 related compounds previously investigated, generated a data base which was used for quantitative structure-activity relationship (qsar) analysis. to this aim, both physico-chemical descriptors and electronic ...19921373821
cloning and properties of a cyanide hydratase gene from the phytopathogenic fungus gloeocercospora sorghi.the cht gene encoding cyanide hydratase (cht, ec 4.2.1.66), which detoxifies hcn and is thought to be important in fungal infection of cyanogenic plants, has been cloned from the phytopathogenic fungus gloeocercospora sorghi. the gene was isolated by screening an expression library of g. sorghi using a cht-specific antibody and using one of the positive cdna clones as a probe in southern hybridization to identify a 3.1 kb psti genomic fragment. this psti fragment expressed cht activity when tran ...19921382413
a mammalian dual specificity protein kinase, nek1, is related to the nima cell cycle regulator and highly expressed in meiotic germ cells.screening of mouse cdna expression libraries with antibodies to phosphotyrosine resulted in repeated isolation of cdnas that encode a novel mammalian protein kinase of 774 amino acids, termed nek1. nek1 contains an n-terminal protein kinase domain which is most similar (42% identity) to the catalytic domain of nima, a protein kinase which controls initiation of mitosis in aspergillus nidulans. in addition, both nek1 and nima have a long, basic c-terminal extension, and are therefore similar in o ...19921382974
reversible inactivation of a foreign gene, hph, during the asexual cycle in neurospora crassa transformants.a plasmid construct carrying the hygromycin phosphotransferase (hph) gene fused to the expression elements of the trpc gene of aspergillus nidulans was used to obtain hygromycin b (hyg)-resistant transformants of neurospora crassa. the plasmid does not have any homology with the n. crassa genome. here we demonstrate that most of the transformants arise from integration of the transforming dna into only one of the nuclei present in the protoplasts. furthermore, in most of the transformants the in ...19921383683
transformation of cochliobolus lunatus with put 720 changes the steroid hydroxylating ability of the fungus.the filamentous fungus cochliobolus lunatus, a known 11 beta-hydroxylator of steroids, was transformed to bleomycin resistance using the heterologous plasmid put 720. this plasmid contains the sh ble gene expressed under the control of the aspergillus nidulans gpd and trpc expression signals. the bleomycin-resistant colonies appeared with a frequency of six per microgram of dna. all colonies were real transformants and no "abortive" growth was observed. in all transformants tested the plasmid mo ...19921384995
an nadp(+)-dependent glycerol dehydrogenase in aspergillus nidulans is inducible by d-galacturonate.in aspergillus nidulans there is an nadp(+)-dependent glycerol dehydrogenase that is specifically induced on transfer to d-galacturonate medium. in contrast to the previously characterised constitutive nadp(+)-dependent glycerol dehydrogenase it has a much broader substrate specificity, having activity as an ethanol dehydrogenase, and is subject to carbon-catabolite repression. in addition to the two nadp(+)-dependent glycerol dehydrogenases, alcohol dehydrogenase i and ii are also present on tr ...19921394511
an upstream activating sequence from the aspergillus nidulans gpda gene.introduction of a previously identified promoter element of the aspergillus nidulans gpda gene (encoding glyceraldehyde-3-phosphate dehydrogenase), the so-called gpd box, into the upstream region of the highly regulated a. nidulans amds gene (encoding acetamidase), significantly increased (up to 30-fold) the expression of the lacz reporter gene fused to these expression signals. this increase was dependent on the orientation of the gpd box and on the site of introduction into the amds upstream r ...19921398125
resolution of chromosomes iii and vi of aspergillus nidulans by pulsed-field gel electrophoresis shows that the penicillin biosynthetic pathway genes pcbab, pcbc, and pende are clustered on chromosome vi (3.0 megabases).an improved electrophoretic molecular karyotype of aspergillus nidulans atcc 28901 has been obtained by contour-clamped electric field gel electrophoresis, which separates seven chromosomal bands and allows resolution of chromosomes iii and vi. the three genes of the penicillin biosynthetic pathway, pcbab, pcbc, and pende, encoding alpha-aminoadipyl-cysteinyl-valine synthetase, isopenicillin n synthase, and isopenicillin n acyltransferase, respectively, are clustered together on a chromosome of ...19921400258
specific binding sites for the activator protein, alcr, in the alca promoter of the ethanol regulon of aspergillus nidulans.alcr is the specific activator of the aspergillus nidulans ethanol-utilization pathway, mediating the induction of its own transcription and that of the structural genes alca and alda, encoding respectively, alcohol dehydrogenase i and aldehyde dehydrogenase. alcr is a dna binding protein in which 6 cysteines are coordinated in a zinc binuclear cluster. this domain was fused to glutathione-s-transferase (gst) and isolated as a gst-alcr(7-58*) fusion protein from escherichia coli. mobility shift ...19921400424
development of a transformation system for the thermophilic fungus talaromyces sp. cl240 based on the use of phleomycin resistance as a dominant selectable marker.a transformation system for the thermophilic cellulolytic fungus talaromyces sp. cl240 has been developed, using the phleomycin resistance gene from streptoalloteichus hindustanus (sh ble) as a dominant selectable marker. the plasmids (pan8-1 and put720) carrying the sh ble gene under the control of the aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter, allowed selection of phleomycin-resistant transformants. a new promoter sequence cloned from chromosomal dna of trich ...19921406595
cmap: contig mapping and analysis package, a relational database for chromosome reconstruction.in the contig mapping and analysis package, cmap, we provide a foundation for reverse genetics by organizing information about dna fragments obtained from an organism's genome into a physical map. the user can store information about a particular segment of dna. this information can be both descriptive, such as any genes contained in a particular dna fragment, or experimental, such as hybridization profiles or restriction digest patterns for comparison with other fragments. the package can then ...19921422880
amino acid alterations in the bena (beta-tubulin) gene of aspergillus nidulans that confer benomyl resistance.we report the cloning and sequencing of 18 mutant alleles of the bena, beta-tubulin gene of aspergillus nidulans that confer resistance to the benzimidazole antifungal, antimicrotubule compounds benomyl, carbendazim, nocodazole, and thiabendazole. in 12 cases, amino acid 6 was changed from histidine to tyrosine or leucine. in four cases, amino acid 198 was changed from glutamic acid to aspartic acid, glutamine, or lysine. in two cases, amino acid 200 was altered from phenylalanine to tyrosine. t ...19921423663
expression of a bacterial aspartase gene in aspergillus nidulans: an efficient system for selecting multicopy transformants.the escherichia coli aspartase gene aspa has been expressed in the fungus aspergillus nidulans using the powerful constitutive gpda promoter and trpc terminator, both from a. nidulans. multiple, but not single, copies of aspa overcome nutritional deficiencies resulting from the loss of catabolic nad-linked glutamate dehydrogenase. they also circumvent certain nutritional deficiencies resulting from loss of the positive-acting regulatory gene product mediating nitrogen metabolite repression. both ...19921423725
transformation of acremonium coenophialum, a protective fungal symbiont of the grass festuca arundinacea.acremonium coenophialum is a mutualistic mycosymbiont and natural agent of biological protection of the widely distributed grass festuca arundinacea (tall fescue). an electroporative transformation system was developed for a. coenophialum. segments of dna 5' to the beta-tubulin gene (tub2) of the closely related ascomycete epichloë typhina, fused to the escherichia coli hph gene encoding hygromycin b phosphotransferase, conferred hygromycin resistance when introduced into a. coenophialum by elec ...19921423727
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