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  • Genetics  (462)
  • Wiley-Blackwell  (462)
  • Oxford University Press
  • Taylor & Francis
  • 1985-1989  (462)
  • 1950-1954
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  • 1
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 93-100 
    ISSN: 0192-253X
    Keywords: heat shock ; phenocopy ; forked ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Heat shock uncovers the recessive forked phenotype when heterozygotes between f36a and wild-type are heated during sensitive periods in pupal development. We call the phenocopy of a mutant in such a heterozygote a heterocopy. The heterocopy in f36a/+ is virtually identical to the mutant phenotype; however, bristles on different parts of the body are affected during different sensitive periods. We discuss the hypothesis that the heat shock acts by affecting expression of the wild-type gene product corresponding to the mutant gene. The sensitive period for heterocopy induction in a specific tissue is proposed to correspond to the normal time of gene expression for the forked gene product in a particular tissue.
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  • 2
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 151-151 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 3
    ISSN: 0192-253X
    Keywords: Dictyostelium discoideum ; revertants of stmF mutants ; cGMP metabolism ; cGMP-specific phosphodiesterase ; suppressor mutations ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: stmF mutants of Dictyostelium discoideum produce long, banded aggregation streams on growth plates and exhibit altered cGMP metabolism. To learn more about the role of cGMP in chemotaxis and the nature of the defect in these mutants, 15 nonstreaming (Stm+) revertants of two stmF mutants were isolated and characterized. Fourteen of the revertants continued to show the elevated cAMP-induced cGMP response and very low cGMP-specific phosphodiesterase (cGPD) activity characteristic of their stmF parents. Parasexual genetic analysis revealed that many of these Stm+ revertants carried phenotypic suppressors unlinked to stmF. One Stm+ revertant, strain HC344, exhibited a low, prolonged cGMP response and relatively high cGPD activity throughout development. To determine whether the elevated cGPD activity in this revertant resulted from increased enzyme production or enhanced enzyme activity, cGPDs were partially purified from the wild-type strain, the stmF parent and revertant HC344, and properties of the enzymes were compared. cGPDs from the stmF mutant and the revertant showed similar differences from the wild-type enzyme in kinetic properties, thermal stability, and sensitivity to certain inhibitors. These results suggest that stmF is the structural gene of the cGPD. In addition, the unusual cGMP response in revertant HC344 appeared to be due to increased production of an altered cGPD.
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  • 4
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 293-293 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 5
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 239-246 
    ISSN: 0192-253X
    Keywords: Drosophila melanogaster ; germ line ; somatic line ; pole cell transplantation ; mosaics ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Drosophila melanogaster mutant fs(1)1304 is an ovary autonomous female sterile mutant that causes abnormal morphology of the egg. Vitellogenesis proceeds at an abnormally slow rate in homozygous females. We have used pole cell transplantation to construct germ line mosaics in order to determine whether the 1304 defect depends upon the genotype of the germ line cells (oocyte or nurse cells) or the somatic line (follicle cells). We have found that the germ line is the primary target tissue where the mutant gene is expressed.
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  • 6
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 269-280 
    ISSN: 0192-253X
    Keywords: UV ; DNA repair ; photoreactivation ; algae ; dark repair ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The response of Volvox to ultraviolet irradiation was analyzed. Young individuals isolated from a synchronous culture were exposed to UV light (120 J/m2) and subjected to variable lenght periods of dark following irradiation. The major effect of the UV treatment was the inability of the gonidia present in the colonies at the time of irradiation to continue and complete the developmental program. Individuals show a heightened sensitivity to UV for a limited period immediately following inversion and are insensitive at other stages of development. The cytotoxic effect of UV during this interval is completely reversed by the immediate exposure to white light and is increased with longer periods of dark treatment prior to exposure to white light. The temporal profile of the sensitivity defines a smooth curve in which the maximal sensitivity occurs three hours after inversion. The response to higher doses of UV (up to 500 J/m2) is a nonlinear increase in cytotoxicity and is disproportionanately greater in those individuals just prior to the period of maximal sensitivity than those later in development. The results suggest that Volvox has at least two pathways for the repair of UV damage and that one of these, the principal dark repair pathway, is temporarily deficient in the gonidia of young individuals.
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  • 7
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 7 (1986) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 8
    ISSN: 0192-253X
    Keywords: tumorous and nontumorous genotypes ; repetitive DNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A single system is presented, where both genetic and epigenetic control of tumor induction can be studied at the same time. This system is offered by the amphidiploid tumorous hybrid Nicotiana glauca × N. langsdorffii, a nontumorous mutant of it and the nontumorous parent species N. glauca and N. langsdorffii. The aim of the present paper is to compare long-term in vitro cultures of tumorous (genetic and habituated), and nontumorous strains, through the characterization of their genomes according to several physico-chemical parameters. The data reported show that both qualitative and quantitative differences in DNA complexity are correlated with the tumorous transformation. Particularly, a high degree of mismatching between the DNAs of the tumorous and nontumorous hybrids and the lack, in the second genotype (nontumorous), of three DNA peaks in Ag+-Cs2SO4 analytical ultracentrifugation profile seem to support the hypothesis, suggested in a previous paper, of the presence, in the nontumorous mutant, of a gross chromosomal rearrangement, probably a deletion. Amplification and underreplication of specific sequences also seemed to be correlated with changes from the normal to the tumorous state, highly repetitive sequences being present in higher amounts in the normal strains and in the habituated N. glauca than in the case of the tumorous hybrid.Finally, DNA bound ion contents were found to be strikingly higher in tumorous than in nontumorous tissues. The results are discussed in the frame of the general hypothesis of high somatic genomic plasticity in plants.
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  • 9
    ISSN: 0192-253X
    Keywords: ecdysteroid ; prothoracic gland ; temperature sensitive ; Drosophila ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The dominant temperature-sensitive mutation L(3)3DTS (DTS-3) in Drosophila melanogaster causes lethality of heterozygotes during the third larval instar at the restrictive temperature (29°C). Temperature-shift experiments revealed two distinct temperature-sensitive periods, with lethal phases during the third larval instar (which may persist for 4 weeks) and during the late pupal stage. At 29°C mutant imaginal discs are unable to evert in situ, but did evert normally if cultured in the presence of exogenous ecdysterone or when implanted into wild-type larval hosts. The only morphologically abnormal tissue present in the lethal larvae is the ring gland, the prothoracic gland being greatly hypertrophied in third instar DTS-3 larvae. Injection of a single wild-type ring gland rescued these mutant larvae, indicating that the mutant gland is functionally, as well as morphologically, abnormal. Finally, the mutant larvae were shown to have less than 10% of the wild-type ecdysteroid levels. These results are all consistent with a proposed lesion in ecdysteroid hormone production in DTS-3 larvae. A comparison with the phenotypes of other “ecdysone-less” mutants is presented.
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  • 10
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 179-197 
    ISSN: 0192-253X
    Keywords: embryonic development ; phenocopies ; heat shock ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Heat shock of pre-adult Drosophila disrupts development and causes phenotypic abnormalities. Type of abnormality depends on developmental stage at time of shock. Defects probably result from disruption of stage specific processes by the heat shock response (which includes reduction of normal mRNA and protein production). This study uses heat shock to study stage specific processes in early development. Short, intense shocks (2-3 min, 42-43°C) are administered to carefully staged embryos within the first 5 h of development. Stage specific defects occur following shock at syncitial blastoderm or later. Abnormal segmentation follows shock at syncitial or cellular blastoderm. Segmentation is also disrupted by shocks 1 h after the onset of gastrulation, but not by shocks at the onset of gastrulation. Segmentation defects include phenocopies of pair rule mutants, which lack parts of alternate segments. Defective shortening of the germ band is common following shock at the onset of gastrulation. Germ band shortening normally occurs several hours after the time of shock; thus heat shock specifically affects control of a later developmental process. Development does not simply cease at the time of the distrupted process; rather a specific step in the developmental sequence is omitted or altered. Stage specific defects do not occur following pre-blastoderm shock. Pre-blastoderm eggs have few or no normal processes controlled by transcription, and poor ability to induce the heat shock response. This suggests that stage specific defects require disruption of transcription controlled processes. Pre-blastoderm eggs survive a 3-min shock less well than older eggs. The ability of older eggs to induce the heat shock response probably enhances survival. The mutant hairy was also investigated. Extreme alleles show a striking pair rule phenotype, while a weak allele does not. Heat shock of animals heterozygous or homozygous for the weak allele at blastoderm specifically increases the frequency of the extreme hairy phenotype. Thus heat shock may disrupt the same developmental process as is altered by the mutation.
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  • 11
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 257-268 
    ISSN: 0192-253X
    Keywords: Drosophila hydei ; cell death ; imaginal discs ; wing reduction ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cell death and its effect on wing size have been described in some wing mutants of Drosophila hydei. Dead cells in the imaginal discs were localized by Nile-bule and acridine-orange staining. Various Notch (N) alleles, the mutation Costal-nick (Cnk) and the compound N/Cnk show characteristic patterns of cell death in the imaginal wing disc. Some but not all of the structural features of the adult wing can be related to the site of cell death during larval stages. In NAx types, extensive cell death is followed by regenerative growth, invalidating a simple relation between size of the disk and size of the wing. In Nts/Cnk cell death and wing morphology depend on the breeding temperature. From temperature experiments we conclude that cell death starts between day 4 and 5 after egg laying and can be induced by a shift to the restrictive temperature during the critical phase. Patterns of wing incisions and cell death in Nts/Cnk genotypes seem not to be delimited by any of the known compartment boundaries.
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  • 12
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 6 (1985), S. 295-296 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 13
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 7 (1986), S. 65-73 
    ISSN: 0192-253X
    Keywords: long interspersed repeated DNA ; demethylation ; myeloma cells ; aging ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sequences of DNA that hybridize on Southern blots with cloned EcoR1 1.3 kb (ER1) of long interspersed repeated sequence (L1Md) of mouse have been examined in genomic DNA of neonatal mice, livers and brains of adult mice (3, 10, 27, and 30 mo old), and the solid myeloma tumor MOPC-315. The isoschizomers Hpa II (CCGG or mCCGG) and Msp I (CCGG or CmCGG) were used to assess methylation. We found that the L1Md sequence is fully methylated in young animals but demethylated in myeloma. Demethylation of L1Md sequence also occurred in aged animals. By scanning the autoradiogram, we found that approximately 8% of the 104-105 copies have been demethylated in 27-mo-old liver.
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  • 14
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 7 (1986), S. 117-117 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 15
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 5 (1989), S. S339 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 16
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 1 (1985), S. 25-38 
    ISSN: 0749-503X
    Keywords: Cyclic AMP ; phosphoprotein phosphatase ; protein kinase ; suppressor ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ppd1 mutant of yeast, Saccharomyces cerevisiae, was isolated as a suppressor of the cyr2 mutation which caused alteration of the catalytic subunit of cAMP-dependent protein kinase. Three peaks of phosphoprotein phosphatase activity (peak I, II and III) were identified by DEAE-Sephacel chromatography of crude extracts of the wild-type strain. The ppd1 mutant was deficient in peak III phosphoprotein phosphatase activity. The peak III enzyme efficiently utilized the phosphorylated forms of NAD-dependent glutamate dehydrogenase and trehalase as substrate. The ppd1 mutation did not suppress the cyr1, CYR3 or ras1 ras2 mutations. The ppd1 locus was located on chromosome II and had identical characteristcs with glc1. The ppd1 mutation suppressed the G1 arrest caused by nutritional limitation, but maintained sensitivity to mating pheromone. In diploids homozygous for the ppd1 mutation, no premeiotic DNA replication and commitment to intragenic recombination occurred and no spores were formed, suggesting that the accumulation of phosphorylated proteins in the absence of one of the phosphoprotein phosphatases is required for mitosis but not for the initiation of meiosis.
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  • 17
    ISSN: 0749-503X
    Keywords: Heat shock ; translational control ; heterologous gene expression ; Saccharomyces ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Plasmid pPW229, containing the 2·25 kilobase transcribed sequence for the 70 000 Dalton heat shock protein of Drosophila,1 was integrated into plasmid CV13 and used to transform Saccharomyces cerevisiae. Upon a heat shock, at 41°C for 20 min, a new 70 000 Dalton protein appeared in the transformants. This protein was not detected in transformants grown at 23°C, nor in transfromants carrying the hybrid plasmid from which the structural gene for the 70 000 Dalton protein had been deleted. RNA was isolated from transromants grown at 23°C and from transformants heat shocked at 41°C. RNA complementary to the Drosophila heat shock gene was present in the transformants, grown either at 23°C or heat shocked. No complementary RNA was detected in yeast cells transformed with the hybrid plasmid from which the structural gene had been deleted. The Drosophila heat shock gene in yeast appears to be transcribed constitutively but translated only under heat shock conditions.
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  • 18
    ISSN: 0749-503X
    Keywords: Apomictic sporulation ; meiosis restoration ; nucleo-mitochondrion interaction ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In an apomitic strain of Saccharomyces cerevisiae (ATCC 4117-H2) which undergoes a single nuclear division during sporulation and consequently forms asci containing two uninucleate diploid spores, a study was undertaken to investigate the effects of cultivation in three presporulation media (YPA; YNB; SMM) on nuclear division and ascoporogenesis in sporulation medium. Comparison of effects of presporulation culture in these media on the number of spores formed per ascus showed that a marked induction (30 ± 4·3 per cent) of three- and four-spored asci could occur in sporulation medium following cultivation in a defined YNB medium supplemented with a 1 per cent solution of vitamins and containing decreased ammonium sulphate and increased glucose levels. Experiments in which the concentrations of glucose and of ammonium sulphate were varied simultaneously indicated that the initial presporulation carbon to nitrogen source ratio is an important factor in determining tetrad formation in sporulation medium. Nuclear staining demonstrated two classes of asci: binucleate (one- and two-spored) and tetranucleate (three- and four-spored). Genetic evidence and data concerning effects of inclusion in sporulation medium of a meiotic inhibitor (glucose) indicated spores in tetrads were haploid rather than diploid. This ability to condition a significant number of cells for meiotic rather than apomictic differentiation made possible investigation of effects of mitochondrial inhibitors on both developmental processes simultaneously. It was found possible to selectively inhibit meiotic development by inclusion in sporulation medium of appropriate concentrations of specific inhibitors. Moreover, the data suggest meiotic sporulation is more strictly dependent than apomictic sporulation on mitochondrial function.
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  • 19
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 2 (1986), S. S281 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 20
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 3 (1987), S. 5-9 
    ISSN: 0749-503X
    Keywords: Schizosaccharomyces pombe ; sterile mutants ; ste genes ; protoplast fusion ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In previous experiments of Girgsdies (1982), eight sterile (ste) mutants of Schizosaccharomyces pombe did not sporulate when fused with h+ or h- protoplasts. We succeeded in achieving sporulation with these mutants. Two hitherto unknown ste genes, ste7 and ste8, were found.
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  • 21
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 3 (1987), S. 1-4 
    ISSN: 0749-503X
    Keywords: Cell cycle ; sporulation ; meiosis ; nuclear division ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cell division age dependency of sporulation was investigated in a diploid strain of Saccharomyces cerevisiae (19el) which undergoes a single equational nuclear division during sporulation with consequent formation of asci containing two uninucleate diploid spores (apomictic dyads). Under modified nutritional conditions which partially restore meiosis and hence normal tetrad formation, newly formed (age 0) daughter cells were observed to be capable of formation of apomictic dyads but not of meiotic tetrads. Even under conditions in which only apomictic dyads developed, approximately 20% of the asci resulted from differentiation of newborn ‘inexperienced’ cells. Thus, the data indicated production of at least one bud to be a prerequisite for meiosis but not for apomixis; however, occurrence of at least one complete mitotic cell division cycle was evidently insufficient for the morphogenetic switch from diploid to haploid spore formation, since older cells bearing several bud scars often underwent apomictic dyad development, and some produced no spores.
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  • 22
    ISSN: 0749-503X
    Keywords: Yeast protein map ; carbon metabolism machinery ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using a modification of the basic two-dimensional polyacrylamide gel electrophoresis technique, we have undertaken a systematic identification of the polypeptides of the protein map of Saccharomyces cerevisiae corresponding to components of the carbon metabolism machinery. To the previous location of nine glycolytic enzyme polypeptides on the yeast protein map we add the location of 23 polypeptides. Ten of them were identified as corresponding to cytoplasmic enzymes of the carbon metabolism machinery and 13 were characterized as mitochondrial proteins. The criteria used to establish the identification of these polypeptides spots include migration with purified proteins, immunodetection, overproduction by plasmid-carrying strains and physiological behaviour.
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  • 23
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 3 (1987) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 24
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    Yeast 3 (1987), S. 263-270 
    ISSN: 0749-503X
    Keywords: Lodderomyces elongisporus ; Rhodotorula gracilis ; Saccharomyces cerevisiae ; accumulation ratio ; membrane transport ; suspension density ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The previously described effect of cell suspension density on metabolic and transport phenomena in yeast, apparently caused by inhibition by dissolved carbon dioxide, is also observed with the accumulation ratio of both sugars and amino acids where not only a kinetic but also an energetic factor comes into play. Unlike all previously measured metabolic and transport parameters, the dependence of the accumulation ratio on suspension density is not monotonic but shows a pronounced maximum in the range of 4-8 mg dry wt/ml, depending on yeast species and on cultivation conditions. In Rhodotorula gracilis and in Lodderomyces elongisporus it is not due to CO2 but is semiquantitatively related to the proton-motive force across the plasma membrane as well as to the intracellular ATP content. It is observed both in oxygen and in argon, over a wide range of pH values and of temperatures, but it is suppressed by metabolic inhibitors. It is expressed only in a range of transported solute concentrations between about 0·1 and 10 mM.
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  • 25
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 4 (1988), S. 1-15 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 26
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    Yeast 4 (1988), S. 27-40 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 27
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    Yeast 4 (1988), S. 17-26 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 28
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    Yeast 4 (1988), S. 41-46 
    ISSN: 0749-503X
    Keywords: Isocitrate lyase ; purification ; Catabolite inactivation ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Isocitrate lyase purified to homogeneity from Saccharomyces cerevisiae was composed of four identical subunits with a molecular mass 75 K Da. The enzyme was most active at pH 7.0 in the presence of 5 mM-Mg2+. The Km value for threo-Ds-isocitrate was 1.4 mM. Isocitrate lyase was shown to be thermostable at 50°C for 60 min at a high salt concentration, but rapidly lost activity at -20°C or by dialysis.
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  • 29
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    Yeast 4 (1988), S. 83-83 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 30
    ISSN: 0749-503X
    Keywords: Bovine leukemia virus ; PH05 ; PGK ; tumor virus ; Saccharomyces cerevisiae ; viral antigens ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: DNA sequences of the envelop (env) gene of the bovine leukemia virus (BLV) were expressed in the yeast saccharomyces cerevisiae. Two yeast promoters, the responsible PH05 promoter and the constitutive PGK promoter, were used to construct four expression plasmids either a sequence of the surface antigen gp51 or a (gp51 + gp30) sequence.The expressed hetrologous gene products were characterized by Western blot analysis and competitive radio-immunoassay. By means of Northern blot analysis the steady-state level of env-specific mRNA was analysed.The highest expression rate was obtained from recombinant plasmid YEpSG 94 comprising a gp51 sequence - a 630 base pair fragment containing 70% of the gp51 but lacking the N terminus - as well as the PH05 promoter including PH05 signal sequence and the PH05 terminator. The recombinant gp51 was partially lycosylated but the PH05 signal peptide did not seem to be cleaved off. No immunoreactive material could be found in the periplasm or in the culture medium.By means of monoclonal antibodies directed against eight different epitopes of viral gp51, all for sequential antigenic determinants were detected in the AH 216(YEpSG 94) expression product.
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  • 31
    ISSN: 0749-503X
    Keywords: Yeast ; Ribosomes ; Kluyveromyces ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In an adenine-requiring mutant strain of the yeast, Kluyveromyces lactis the intracellular content of ATP is one-third to one-fifth that in a protophic wild strain under growing conditions. The quantitatives difference becomes rather small in resisting stationary-phase cells. Temporary changes in the two-dimensional protein patterns of mutant ribosomes occur when the ATP content during the transition phase of growth. The transfer of exponentially growing cells to a synthetic complete medium void of adenine induces the same changes in mutant ribosomes within several hours. Identification of robosomal proteins by two-dimensional gel electrophoresis indicated all changeable proteins (at least five proteins) to belong to 40S ribosomal subunits. The mutant ribosomes prepared from the transitio-phase cells have much lower activity (below 60%) for poly(U)-directed polyphenylalanine synthesis than those in exponentially growing or resisting stationary-phase cells. Thus, changes in ribosomal components associated with the differences in ribosomes activity in a cell-free system were noted in the adenylate-deprived cells of K. lactix.
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  • 32
    ISSN: 0749-503X
    Keywords: Kluyveromyces lactis ; killer DNA plasmid ; gene cloning ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The killer system of Kluveromyces lactis is associated with two linear DNA plasmids, pGKL1 and pGKL2. The killer toxin and the immunity determinant are coded for by pGKL1. Mutations which we have named KEX1. The KEX1 gene of K. lactis has been cloned by complementation of kex1 mutations by using a recombinant plasmid pool containing the entire Kluyveromyces lactis genome, on a multicopy plasmid KEp6, which contains the Saccharomyces cerevisiae URA3 gene as a marker. Genetic analyses of strains carrying a distrupted kex1 allele demonstrated that the cloned DNA corresponded to the KEX1 gene. The cloned KEX1 gene of K. lactis has low but significant sequence homology with the KEX2 gene of Saccharomyces cerevisiae. In vivo complementation of the kex1 mutations of K. lactis by the KEX2 gene of S. cerevisiae, and complementation of the kex2 mutations of S. Cerevisiae by the KEX1 gene of K. lactis, demonstrated that KEX1 of K. Lactis is functionally related to the KEX2 gene of S. cerevisiae. K. lactis diploids homozygous for kex1 are deficient for sporulation.
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  • 33
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    Yeast 4 (1988) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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  • 34
    ISSN: 0749-503X
    Keywords: Yeasts ; dihydroxyacetone ; acetoin ; diacetyl ; acetol ; methylglyoxal, acetone ; glycerol ; 1,2-propanediol ; 2,3-butanediol ; dehydrogenase ; reductase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Hansenula polymorpha CBS 4732 grown on a variety of substrates contained very high activities of enzymes catalyzing the NADH-linked reduction of dihydroxyacetone, acetoin, diacetyl, acetol, methylglyoxal and acetone. The enzymes catalyzing these reductions have been purified and their kinetic properties are described. Three different enzymes were found responsible for the above-mentioned activities, namely: (1) dihydroxyacetone reductase; (2) acetone reductase; and (3) alcohol dehydrogenase.So far, the physiological function of dihydroxyacetone reductase and acetone reductase is obscure. The kinetic properties of dihydroxyacetone reductase and the regulation of the synthesis of this enzyme suggest that it does not function as a glycerol dehydrogenase.
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  • 35
    ISSN: 0749-503X
    Keywords: Dihydroxyacetone reductase ; 2,3-butanediol dehydrogenase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Candida utilis CBS 621 contained four different enzymes capable of reducing carbonyl compounds such as dihydroxyacetone, acetoin, diacetyl, acetol, methylglyoxal and acetone, namely alcohol dehydrogenase, acetone reductase, dihydroxyacetone reductase and 2,3-butanediol dehydrogenase. The dihydroxyacetone reductase of C. utilis did not oxidize glycerol, thus providing evidence that this enzyme cannot function as a glycerol-2-dehydrogenase during growth of the yeast on glycerol. This enzyme may, however, play a role in the assimilation of 2,3-butanediol by C. utilis. The organism also contained a separate 2,3-butanediol dehydrogenase which was unable to reduce dihydroxyacetone. Both dihydroxyacetone reductase and 2,3-butanediol dehydrogenase were present at very high activities during growth of C. utilis on a variety of substrates, including 2,3-butanediol.
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  • 36
    ISSN: 0749-503X
    Keywords: Single-cell proteins ; Saccharomyces cerevisiae ; fragile mutants ; srb1 ; lysis ; polyploids ; protein extracts ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A series of prototrophic fragile strains of different ploidy (2n, 3n and 4n) has been genetically constructed on the basis ofhalopoid srb1 containing segregants of the fragile Saccharomyces cerevisiae mutant VY 1160. The strains have been characterized by several criteria. In regard to generation time, biomass yield, and nucleic acids content of the cells, the tetraploid srb1 homozygous hybrid is indistinguishable from an industrial strain of S. cerevisiae. However, it is characterized by a higher protein content. Unlikely any other laboratory or industrial strains, the original mutant and these hybrids possess an ability for lysis upon suspension in hypotonic solutions. The dependence of the percentge of lysed cells on the growth phase and concentration of osmotic stabilizer in the medium has been investigated. The quantity of proteins in the soluble fractions obtained after lysis of these strains by osmotic shock has been determined. These hybrids can be considered as a potential industrial source of potentials for nutritional purposes.
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  • 37
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    Yeast 4 (1988) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 38
    ISSN: 0749-503X
    Keywords: Furctose-1 ; 6-biophosphatase ; Saccharomyces cerevisiae ; specificity of phosphatases ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Enzymatic dephosphorylation of the phosphorylated forms of five different yeast enzymes has been studied: fructose-1,6-bisphosphatase, glycogen phosphorylase, neutral trehalase, NAD-glutamate dehydrogenase and 6-phosphofructo-2-kinase. Phosphorylated fructose-1,6-bisphosphatated 6-phosphofructo-2-kinase were present in extracts of starved yeast cells which had been incubated for 10 min with glucose. Phosphorylated glycogen phosphorylase, neutral trehalase and NAD-glutamate dehydrogenase were obtained by incubation of yeast extract with ATP, cycle AMP and Mg2+. After incubation with commercially available preparations of alkaline phosphatase, all five phosphorylated enzymes studied showed the changes in catalytic activity that would be expected as a consequence of dephosphorylation. The recently purified yeast enzyme which dephosphorylates phosphorylated fructose-1,6-bisophosphatase (Horn and Holzer (1987)) however, was found to be active only with the phosphorylated fructose-1,6-bisphosphatase, but not with the other four phosphorylated enzymes studied. By contrast, a crude extract from yeast showed dephosphorylating activity towards all five substrates. Substrate specificity with the five phosphorylated enzymes studied of different phosphoprotein phosphatases from yeast prepared by other is discussed.
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  • 39
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    Yeast 4 (1988), S. 241-247 
    ISSN: 0749-503X
    Keywords: Membrane transport ; fragile muatnt ; H+ extrusion ; spontaneous acidfication ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Transport properties of the osomotically fragile strain VY1160 of saccharomyces cerevisiae were compared with those of the parent S288c strain. Mediated diffusion of 6-deoxy-D-glucose was practically unaffected; membrane-potential dependent transport of D-glucosamine was very much depressed in the fragile strain. The H+ -driven transport of L-lysine and Lproline, as well as that of the hitherto uninvestigated D-glucose-6-phosphate, were also very depressed. 2-Deoxy-D-glucose transport displayed slightly different kinetic parameters. Primary H+ extrusion by the plasma membrane H-ATPase was not diminished althpough the ATP-splitting activity was depressed by about 50%. The overall proton-motive force (pmf) of the fragile mutant at pH 5.5 was only m V while in the parent strain it was 108 m V. In parallel with this, spontaneous acidfication of the external medium to stimulate (a CO2-associated event) was only about 2% of that in the parent strain. The defect in his, together with the inability to stimulate transport protein synthesis by glucose, may account for the generally poorer transport performance of the fragile mutant.
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  • 40
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    Yeast 4 (1988), S. 249-255 
    ISSN: 0749-503X
    Keywords: Brettanomyces ; custers effect ; glycosis ; organic hydrogen acceptors ; mass spectrometry ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The yeast Brettanomyces anomalus showed the Custers effect in that under strictly anaerobic conditions, in the presence of glucose, CO2 production was negligble. CO2 production was stimulated by mixing anaerobic cell suspensions with an aerated glucose solution in astopped-flow cell. Glycolytic CO2 production continued even after oxygen exhaustion. Studies using an open reaction vessel showed that the rate of glycolytic CO2 production could be increased to a maximum level by exposing the anaerobic cell suspension to brief pulses of O2. A cell suspension CO2 at a maximal rate demonstrated the Pasteur effect on switching the mobile gas to a mixture conatining oxygen (5.05 KPa). In contrast to glycolytic CO2 production in vivo nicotinamide pool responded rapidly to changes in oxygen concentration. The addition of acetaldehyde, acetone, or 3-hydroxy-butan-2-one led to a temprorary production of CO2 at an initial rate depending on the concentration of substance added according to the Michaelis-Menten equation. The maximal rates were equal with all three substances, whereas tha apparent Km values were different. The total amount of CO2 produced was 22-fold greater than the amount of acetaldehyde added. Added organic hydrogen acceptors modulated the intracellular reedox balance of B. anomalus under conditions. These results are discussed in relation to the current hypothesis of the Custers effect.
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  • 41
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; yeast ; chromosomes ; cell division ; mitosis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have identified four new genetic loci: CHL2 (on chromosome XII) CHL3 (on chromosomes XII); CHL4 (on chrosomes IV), and CHL5 (on chromosomes IX), controlling mitotic transmission of yeast chromosomes. The frequency of loss of chromosomes is 10-100-fold in chl5, chl2, chl3 and chl4 mutants than observed in wild-type strains. The mutants also unstable maintenance of artifcial circular minichromosomes with various chromosomal replicators (ARS) and one of the concentrations loci (CEN3, CEN4, CEN5, or CEN6). The instability of minichrosomes in the chl5, chl2, and chl4 mutants id due to the loss of minichromosomes in mitosis (1 : 0 segregation). In the chl3 mutant the instability of artificial minichromosomes is due to nondisjunction (2 : 0 segregation). The CHL3 gene therfre appears to affect the segregation of chromosomes during cell division.
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  • 42
    ISSN: 0749-503X
    Keywords: DNA sequence ; ras related ; membrane localization ; palmitoylation ; C-terminal modification ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ras protein represent a unique example of membrane proteins which apparently do not utilize the secretory pathway for their membrane localization. Instead, it is belived that palmaitic acid, covalently attached to the protein, acts as an anchor to the membranes. Recent identification of yeast mutants defective in the processing of the ras proteins has provideda novel approach for defining these biosynthetic process. We report here the charcterization of yeast DPR1, a gene essential for the processing of the ras proteins. The sequence of the gene indicates that it encodes a protein of 431 amino acids which contains no significant homology with any known proteins. It is a relatively hydrophilic protein of cysteine. The DPR1 gene product product has been identified in a cell-free translation system as a proteinhaving an apparent molecular weight of 43 hd. This represents the first step in the translation system as a protein having an apparent molecular weight of 43 kd. This represents the first step in the investigation of a novel protein-processing pathway, one that id distinct from the secretory pathway.
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  • 43
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    Yeast 4 (1988), S. ix 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 44
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    Yeast 4 (1988), S. S181 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 45
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    Yeast 4 (1988), S. S191 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 46
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    Yeast 4 (1988), S. S207 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 47
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    Yeast 4 (1988), S. S243 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 48
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    Yeast 4 (1988), S. S269 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 49
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    Yeast 4 (1988), S. S287 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 50
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    Yeast 5 (1989), S. ii 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 51
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    Yeast 5 (1989), S. 1-10 
    ISSN: 0749-503X
    Keywords: Yeast ; genome size ; orthogonal-field-alternation gel electrophoresis ; mitochondrial DNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using an improved procedure of pulsed field gel electrophoresis, yeast chromosomes were separated over a wide range of molecular size (250-4000 kbp) on single gels. The chromosomal DNA patterns of all the species belonging to the genus Kluyveromyces were examined. Within the species K. marxianus, the varieties lactis, drosophilarum and vanudenii showed closely related patterns; very different from them, the varieties bulgaricus and marxianus were related to each other, forming a distinct group; the strains commonly called ‘K. lactis’ and ‘K. fragilis’ were unambiguously different from each other in chromosome patterns. These differences were correlated with the presence of characteristic repetitive sequence elements in the mitochondrial DNA of the former group and not in the latter. Analysis of Candida macedoniensis, which had been considered to be an anamorph of K. marxianus var. marxianus, showed that these two yeast species were indeed similar in chromosome patterns and in mitochondrial DNA restriction patterns.
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  • 52
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    Yeast 5 (1989), S. S471 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 53
    ISSN: 0749-503X
    Keywords: Killer ; virus-like particles ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: L-A-E double-stranded RNA (dsRNA), when introduced into cells carrying L-A-H and M2 dsRNAs, does not eliminate the L-A-H dsRNA, but (i) L-A-E does lower the copy number of L-A-H dramatically and (ii) L-A-E eliminates M2 dsRNA from the cell. That these two effects of L-A-E are related is shown by the fact that mutants of a strain carrying L-A-H and M2 selected for their resistance to exclusion of M2 by L-A-E [effect (ii)] have an altered L-A-H whose copy number is not lowered by L-A-E [effect (i)]. Although the L-A in K1 strains (L-A-HN in all cases examined) differs significantly both genetically and physically from the L-A in the K2 strain studied (L-A-H), the L-A-HN from the K1 strains can maintain M2 dsRNA, and the L-A-H from the K2 strains can maintain M1 dsRNA.
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  • 54
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    Yeast 1 (1985), S. 82-82 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 55
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    Yeast 1 (1985) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 56
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    Keywords: Life and Medical Sciences ; Genetics
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  • 57
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    Yeast 1 (1985), S. 159-171 
    ISSN: 0749-503X
    Keywords: PET18 ; temperature sensitive growth ; killer ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The basis of pleiotropy shown by the pet18 mutants of Saccharomyces cerevisiae (rho-0, KIL-0 and temperature sensitive growth) was examined by cloning the fragment which complements the defect in growth at 37°C of the pet18 mutants. the cloned DNA could complement the defect in the maintenance of the killer plasmid but did not give the cell the ability to maintain mitochondrial DNA. Sequence analysis of the cloned DNA revealed the presence of four open reading frames, at least two of which are necessary for the complementation activity. By using the cloned DNA as a probe, we found that two independent pet18 mutants have a deletion covering the entire sequence contained in the probe. From these results we predict that the traits of the pet18 mutants that concern temperature sensitivity and killer of the pet18 mutants are controlled by a separate gene(s) from that which participates in the maintenance of mitochondrial DNA.
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  • 58
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    Yeast 2 (1986) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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  • 59
    ISSN: 0749-503X
    Keywords: Amine oxidase ; yeast ; methylamine ; n-butylamine ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Under conditions known to separate methylamine oxidase from benzylamine oxidase in other yeast strains, only a single oxidase could be detected in Sporobolomyces albo-rubescens. This occurred irrespective of whether methylamine or n-butylamine was the nitrogen source for growth. The oxidase did not attack benzylamine. It was concluded that this organism can only produce a methylamine oxidase. The enzyme was purified to 90% homogeneity and found to have properties significantly a methylamine oxidases previously characterised. It lost only 40% of its activity in 30 min at 45°C, whereas methylamine oxidase previously described had half-lives of from 2 to 9 min at 45°C. It showed also a lower activity with short chain 1-aminoalkanes and a higher activity with longer chain 1-aminoalkanes than other methylamine oxidases, and had a significantly smaller subunit molecular weight (57 000 compared with 80 000).
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  • 60
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    Yeast 2 (1986), S. 77-85 
    ISSN: 0749-503X
    Keywords: Vanadium metabolism ; magnetic resonance spectroscopy ; respiratory-deficient strains ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Vanadium metabolism was studied in a wild type and respiratory-deficient strains of S. cerevisiae. Inhibition of growth by vanadate [V(+5)], vanadate accumulation, and conversion of medium vanadate [V(+5)] to both cell-associated and medium vanadyl [V(+4)] and vanadate [V(+5)] were compared. The growth of both the parental and respiratory-deficient strains was inhibited by vanadate at concentrations greater than or equal to 1 mM. Both parental and respiratory-deficient strains accumulated vanadate and converted medium vanadate to cellular vanadyl as detected using electron spin resonance (ESR). The accumulation of cell-associated vanadyl was correlated with the loss of medium vanadate in both strains using a chemical assay. In contrast, the respiratory-deficient strain showed a greater amount of a cell-associated vanadate compound, as detected with vanadium-51 nuclear magnetic resonance (51V-NMR), than the wild strain or a representative respiratory-competent vanadate-resistant mutant. These data imply that mitochondrial function may be directly involved in vanadium metabolism.
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  • 61
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    Yeast 2 (1986), S. 93-100 
    ISSN: 0749-503X
    Keywords: pH ; weak acid ; synergism ; growth ; Zygosaccharomyces ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In completely randomised factorial experiments, individual and synergistic effects of pH, benzoic acid and sorbic acid on the growth rate of the yeast Zygosaccharomyes bailii were determined, and expressed in polynomial equations. Synergism between benzoic and sorbic acid was pH dependent. A distinct effect of the anionic form of benzoic acid on doubling time was demonstrated by experiments in which concentrations of benzoic acid and benzoate were varied. The resultant polynomial equation showed that both species act synergistically.
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  • 62
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    Yeast 2 (1986), S. 101-108 
    ISSN: 0749-503X
    Keywords: L-Arabinose ; D-xylose ; UDP-glucose 4-epimerase ; inactivation ; Saccharomyces ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In a previous paper (Cármenes et al., 1984) we reported that UDP-glucose 4-epimerase from Saccharomyces was inactivated both in vitro (Crude extracts) by L-arabinose or D-xylose. In this paper, we reported that pure epimerase requires the presence of UMP or UDP to be inactivated by sugars and that the inactivation is due to the reduction of the epimerase NAD+, which is essential for epimerase activity. The inactivation rate is directly proportional to epimerase and sugar concentrations and hyperbolically proportional to UMP concentration. In situ experiments made with permeabilized cells showed that epimerase is inactivated in the same way when it is inside the cell. In vivo studies showed that epimerase is inactivated to a smaller extent when 1% Dgalactose is present in the culture medium than when 1% ethanol is the main carbon source.
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  • 63
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    Yeast 2 (1986), S. 109-115 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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    Topics: Biology
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  • 64
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    Yeast 2 (1986), S. 117-121 
    ISSN: 0749-503X
    Keywords: Hexose monophosphate pathway ; NADPH ; radiorespirometry ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A comparative radiorespirometric study of glucos emetabolism in glucose-limited chemostat cultures of Saccharomyces cerevisiae, Candida utilis and Rhodosporidium toruloides was performed in an attempt to estimate the contribution of the hexose monophosphate (HMP) pathway to glucose metabolism. Radioactively labelled glucose was administered directly to the cultures in a constant substrate feed, without disturbance of the steady state. The 14CO2 yields from [1-14C]- and [6-14C]-glucose demonstrated that the HMP pathway activities for the three yeasts were very similar. Furthermore, a quantitative analysis of results indicated that the HMP pathway activities were close to the theoretical minimum needed to cover the NADPH requirement for biomass formation.
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    Yeast 2 (1986), S. 141-142 
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    Keywords: Life and Medical Sciences ; Genetics
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    Yeast 2 (1986), S. 129-139 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; regulation ; pho ; pho80 ; CEN15 ; nucleotide sequence ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The PHO80 gene, which is one of the regulatory genes exerting negative control in the pho system of Saccharomyces cerevisiae, was cloned. The 1·8 kb DNA fragment carrying the PHO80 gene was sequenced and one open reading frame large enough to encode 293 amino acids was found in the sequence. Northern blot analysis of poly(A)+-RNA isolated from cells grown under repressed and derepressed conditions revealed that (i) the size of the PHO80 message was around 1·4 kb, (ii) the expression of the PHO80 gene was not affected by the presence or absence or absence of inorganic phosphate in the medium, and (iii) the expression of the PHO80 gene was not affected by pho2, pho4, pho81, or by pho80 itself. a centromere sequence was found downstream of the PHO80 coding region.
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    Yeast 2 (1986), S. S301 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 68
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    Yeast 2 (1986), S. S321 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 69
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    Yeast 2 (1986), S. S341 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 70
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    Yeast 2 (1986), S. S361 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 71
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    Yeast 2 (1986), S. S381 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 72
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    Yeast 2 (1986), S. S401 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 73
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    Yeast 3 (1987), S. 43-49 
    ISSN: 0749-503X
    Keywords: Heterologous gene expression ; Hepatitis B ; protein estimation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Purified recombinant hepatitis B surface antigen separated on polyacrylamide gels in the presence of sodium dodecyl sulphate has a very low staining index with Coomassie blue relative to a number of standard proteins. In contrast the protein stains better than average with silver nitrate. This property has been used to develop a semi-quantitative method of estimation of recombinant surface antigen in extracts of Saccharomyces cerevisiae producing this protein. The method can be used to follow purification protocols. It is quick, simple and since it measures the surface antigen biochemically, is independent of the aggregation state or conformation of the protein, a factor which can affect enzyme-linked immunoassays which rely on antigen-antibody interactions.
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  • 74
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    Yeast 3 (1987), S. 33-42 
    ISSN: 0749-503X
    Keywords: Yeast ; acid phosphatase ; gene regulation ; upstream activating sequences ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: To identify the sequences involved in the regulation of the yeast acid phosphatase gene (PHO5) we constructed a series of hybrid promoters. Increasing lengths of 5′-flanking sequences of the PHO5 gene were placed in front of the TATA-box of constitutively expressed acid phosphatase gene (PHO3).The PHO5/PHO3 promoter constructions were used to replace the entire PHO5, PHO3 gene cluster on chromosome II. Depending on the length of PHO5 5′-flanking sequences present the PHO3 gene driven by the hybrid promoter could now be derepressed in response to inorganic phosphate (low Pi) exactly as the PHO5 wild type gene. A critical regulatory element was located between position -402 to -351 (upstream from ATG) and sequences further downstream (from -351 to -300) could increase transcriptional activation. The transcription levels of PHO3 were determined by northern blot analysis, under repressed (high Pi) and derepressed (low Pi) conditions which was paralleled by an increase in extra-cellular acid phosphatase activity. Fully regulated promoter hybrids showed a 40-fold induction of mRNA levels, comparable to wild type PHO5 promoter. S1-nuclease protection experiments revealed that the PHO5 5′-flanking sequences, placed in front of PHO3, did not change the PHO3 transcription initiation site/s.
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  • 75
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    Yeast 3 (1987) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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  • 76
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    Yeast 3 (1987), S. 117-129 
    ISSN: 0749-503X
    Keywords: Killer ; virus-like particles ; nucleotides ; pyrophosphatase ; RNA polymerase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The intracellular killer virions of yeast co-purify with an RNA polymerase activity which catalyzes the synthesis of fulllength transcripts of the two viral genomic double-stranded RNA segments. This polymerase utilizes ribonucleoside diphosphates or triphosphates as substrates. The virions have other associated nucleotide-metabolizing enzyme activities, including nucleoside diphosphate kinase, adenosine monophosphate kinase, and nucleoside triphosphate phosphotransferase, an activity which catalyzes the exchange of gamma-phosphate from any ribonucleoside triphosphate with any ribonucleoside or deoxyribonucleoside triphosphate. The purified virions also contain an inorganic pyrophosphatase activity. These enzymes may allow the virus to utilize nucleotide pools distinct from those utilized in host cell transcription.
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  • 77
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    Yeast 3 (1987), S. 131-137 
    ISSN: 0749-503X
    Keywords: Transformation ; Saccharomyces ; plasmid ; DNA uptake ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied the mechanism of DNA transformation of whole yeast cells in Saccharomyces cerevisiae with particular emphasis on the role of the cell wall complex in DNA uptake. Two new aspects of the process have been investigated in order to evaluated its specificity. Such aspects are: (i) effect of monovalent vs. divalent cations during incubation with the transforming DNA and (ii) timing of DNA adsorption and uptake. We found that the specificity for cation requirement is a strain-dependent characteristic influenced by the presence of transforming DNA in the cell suspension. This finding is supported by reports from several laboratories that some yeast strains show mutually exclusive transformability with monovalent vs. divalent cations. While irreversible adsorption of plasmid DNA molecules is induced by both heat shock and polyethylene-glycol(PEG), DNA uptake seems to occur only after the removal of PEG. In the course of this study we have developed a new, alternative method of whole cell DNA transformation with CaCl2 able to transform strains that do not respond to other methods.
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  • 78
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    Yeast 3 (1987), S. 139-148 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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    Topics: Biology
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  • 79
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    Yeast 3 (1987), S. 187-200 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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    Topics: Biology
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  • 80
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; mitochondria ; cAMP-dependent protein kinase ; submitochondrial localization ; topology ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe the identification and submitochondrial localization of four protein kinases and of their target proteins in derepressed yeast mitochondria. The activity of one of the kinases depends on the presence of cyclic AMP (cAMP). It is soluble and localized in the mitochondrial intermembrane space. Its natural target is a polypeptide of 40 kDa molecular mass, which is bound to the inner membrane. Besides this natural target this kinase phosphorylates acidic heterologous proteins, like casein, with high efficiency. The other protein kinases identified so far are cAMP-independent. At least one is localized in the matrix having its natural substrates (49 and 24 kDa) in the same compartment. Two others are firmly bound to the inner membrane phosphorylating target proteins in the inner membrane (52·5 kDa) and in the intermembrane space (17·5 kDa), respectively.
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  • 81
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    Yeast 3 (1987), S. 149-160 
    ISSN: 0749-503X
    Keywords: DNA repair ; RAD2 ; Saccharomyces ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cloned RAD2 gene of S. cerevisiae was tailored into regulatable expression vectors for overexpression of Rad2 protein in E. coli and in yeast. In E. coli both Rad2/β-galactosidase fusion protein and native Rad2 protein are insoluble, but are extractable with 1% Sarkosyl. In yeast some of the overexpressed native Rad2 protein is also insoluble; however, soluble protein is readily detected by immunoblotting with Rad2-specific antibodies. All forms of the protein detected in transformed or untransformed yeast cells and the insoluble species in E. coli migrate in denaturing polyacrylamide gels with an apparent molecular weight considerably larger than the size predicted from the sequence of the RAD2 coding region. This property is not the result of post-translational glycosylation detectable by binding of concanavalin A, or of phosphorylation of the protein. Overexpression of the RAD2 gene is toxic to yeast. Transformed yeast cells grow much more slowly than untransformed controls and when yeast transformants are serially propagated cultures show considerable colony heterogeneity and concomitant selection for rapidly growing variants which express less Rad2 protein. Antisera raised against Rad2/β-galactosidase fusion protein expressed in E. coli do not cross-react with Rad1, Rad3 or Rad10 protein in crude extracts of yeast, nor with purified E. coli UvrA, UvrB or UvrC proteins.
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  • 82
    ISSN: 0749-503X
    Keywords: Cellulases ; Endoglucanases ; Trichoderma reesei ; Saccharomyces cerevisiae ; Cellulolytic yeast ; secretion ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cDNA copies of the two endo-β-1,4-glucanase genes, egl1 and egl3, from the filamentous fungus Trichoderma reesei were expressed in yeast Saccharomyces cerevisiae under the control of the yeast phosphoglycerate kinase gene promoter. Active EGI and EGIII enzyme was produced and secreted by yeast into the growth medium. The recombinant EGI enzyme was larger and more heterogeneous in size than the native enzyme secreted by Trichoderma, due to differences in the extent of N-glycosylation between these two organisms. The morphology of the yeast cells producing EGI or EGIII was clearly different from control strain.
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  • 83
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    Yeast 3 (1987), S. 209-221 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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  • 84
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    Keywords: Life and Medical Sciences ; Genetics
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  • 85
    ISSN: 0749-503X
    Keywords: Pichia pinus ; alcohol oxidase ; catabolite repression ; metabolic regulation ; methanol ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The effect of various carbon compounds on the synthesis of alcohol oxidase in a medium with methanol was studied in the wild type strain of Pichia pinus as well as in gcr1 and ecr1 mutants defective in glucose and ethanol repression of methanol metabolic enzymes, respectively. Compounds repressing the synthesis of alcohol oxidase in the wild type strain were divided into four groups. Repression of alcohol oxidase by compounds of the first group (glucose, fructose, mannose, galactose, L-sorbose and xylose) was impaired only in the gcr1 mutant and that by compounds of the second group (ethanol, acetate, 2-oxoglutarate and erythritol) only in the ecr1 mutant. Repression by compounds of the third group (malate, dihydroxyacetone) was not impaired in both these regulatory mutants and that by compounds of the fourth group (succinate, fumarate, L-arabinose, sorbitol, salicin, xylitol and cellobiose) was partially reduced in both gcr1 and ecr1 strains.Mutation gcr1 causes a significant decrease in phosphofructokinase activity. It also led to a six- to seven-fold increase in intracellular pools of glucose-6-phosphate and fructose-6-phosphate and to a two-fold decrase in the intracellular pool of fructose-1,6-bisphosphate. In ecr1 strains, a decrese in 2-oxoglutarate dehydrogenase activity accompanied by an increae in activities of NAD- and NADP-dependent isocitrate dehydrogenases and NAD- and NADP-dependent glutamate dehydrogenases was demonstrated. The intracellular pool of 2-oxoglutarate was increased 2·5-fold in ecr1 strains. Genes GCR1 and ECR1 are not linked.The mechanisms of catabolite repression of alcohol oxidase in methylotrophic yeasts are discussed.
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  • 86
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    Yeast 4 (1988), S. 85-92 
    ISSN: 0749-503X
    Keywords: Cyt. P450 ; Schizosaccharomyces pombe ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The yeast Schizosacchromyces pombe has been shown to contain a microsomal cytochrome P-450 (cyt. P-450) inducible under conditions of glucose repression. Under these conditions the enzyme has maximal expression of 0.43 nmol g-1 wet wt at the end of the exponential phase of growth. Substrate and inhibitor affinity was examined using studies of spectral changes on binding and revealed a type II spectrum with ketoconazole (Ks = 23 μM) and a type I spectrum with benzo(a)pyrene (Ks = 77 μM). A Km of 112 μM was found in the aryl hydrocarbon hydroxylas assay. These properties show broad comparability with the cyt. P-450 of Saccharomyces cerevisiae.
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  • 87
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    Yeast 4 (1988), S. 159-178 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
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    Topics: Biology
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  • 88
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    Yeast 4 (1988), S. 179-190 
    ISSN: 0749-503X
    Keywords: Zygosaccharomyces ; yeast plasmids ; sequence homology ; plasmid evolution ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genomic DNAs isolated from 420 yeast strains stocked in the Department of Fermentation Technology, Hiroshima university (HUT) were screened fro the presence of a plasmid sequence both as plasmid or in the chromosome. Five DNA samples gave rise to a positive hybridization signal wht 32P-labelled Zygosaccharomyces plasmid pSR1 was used as a probe. Two among these contain hybridzing sequences as plasmids while the other three apparently were chromosomal. Two chromosomal DNA segments of HUT 7195 (Zygosaccharomyces spp.) which hybridized with pSR1 probe were cloned and sequenced. Both DNAs hybridized with a plasmid sequence covering the P gene of pSR1. One of hte tow segments contains a large open reading frame which can encode 410 amino acid residues. The deduced amino acid sequence is closely related with that of the P gene of pSR1. The present finding suggests that there was an interchange(s) of a gene between yeast plasmid(s) and chromosomes.
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  • 89
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    Yeast 4 (1988), S. 199-208 
    ISSN: 0749-503X
    Keywords: Flocculation ; yeast ; agitation ; equilibrium ; mannose ; pH value ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The steady state in yeast flocculation is a dynamic equilibrium between flocculated and dispersed yeast cells. The free cell concentraiton is directly proportional to the total cell concentration and may be expressed as an equilibrium constant. Increased agitation decreases floc size and equlibrium constant whilst increasing floc-surface area and free cell concentration. Values of equilibrium constant are influenced by agitation in a complex relationship probably involving the floc-surface area and floc momentum.Inhibition of flocculation by mannose and low pH is reversible and becomes greater with increased agitation. Both these inhibitions appear consistent with a weakening of flocculent bond strength by these inhibitors.
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  • 90
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    Yeast 4 (1988), S. 235-240 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 91
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    Yeast 4 (1988) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 92
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    Yeast 4 (1988), S. 293-303 
    ISSN: 0749-503X
    Keywords: Hansenula polymorpha ; methylotrophic yeast ; genetic analysis ; methanol mutant ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Techniques are described for the induction, isolation, and characterization of mutants of Hansenula polymorpha. In addition, techniques for controlled passage through the life cycle and genetic analyses, including complementation, tetrad and random spore analysis, have been developed and used to assign mutants to 62 complementation groups. We report that organism conforms to the expected genetics of a homothallic yeast and displays a Mendelian segregation of genes through meiosis. Preliminary mapping data are presented indicating linkage of three genes on a single linkage fragment. Enymatic analysis of methanol-non-utilizing mutants identified one class which is totally deficient in the key assimilatroy enzyme, dihydroxyacetone synthase.
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  • 93
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    Yeast 5 (1989), S. 25-33 
    ISSN: 0749-503X
    Keywords: Secretion ; Saccharomyces cerevisiae ; Golgi apparatus ; protein targetting ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The KEX2 protease (product of the KEX2 gene) functions late in the secretory pathway of Saccharomyces cerevisiae by cleaving the polypeptide chains of prepro-killer toxin and prepro-α-factor at paired basic amino acid residues. The intracellular vesicles containing KEX2 protease sedimented in density gradients to a position distinct from those containing mannosyltransferase I (product of the MNN1 gene), a marker enzyme for the Golgi complex. The recovery of intact compartments containing these enzymes approached 80% after sedimentation. We propose that the KEX2 protease and mannosyltransferase I reside within distinct compartments.
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  • 94
    ISSN: 0749-503X
    Keywords: Yeast ; α-glucosidase ; nucleotide sequence ; expression ; proteinase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Two α-glucosidase (maltase) genes, designated GLUCPI and GLUCPII, have been cloned from an industrial strain of baker's yeast (Saccharomyces cerevisiae) by complementation of a maltase-negative mutant strain. The different genes were identified according to their alternatively expressed isoenzymes PI and PII in transformants after isoelectric focusing and activity staining in separated cell lysates. The gene encoding α-glucosidase PI (GLUCPI), which was not present in laboratory strains of S. carlsbergensis with a defined MAL1, 2, 3, 4 or 6 locus, was sequenced and compared with the recently published MAL6S gene. This comparison revealed single amino acid deviations at three positions in the predicted polypeptide sequence. In addition, the divergent promoter region of GLUCPI differed from MAL6S by a triple repeated 147-bp DNA segment. Maltose induction and glucose repression of α-glucosidase PI were not affected by the deletion of the repeated DNA segment. However, the absolute expression of α-glucosidase PI increased two- to four-fold. In addition, a two-fold increase in the maltase synthesis occurred when the cloned positive regulator gene MAL2-8cp was on the same plasmid. Furthermore, stability of the α-glucosidase in cultures in the stationary growth phase was greatly enhanced using a host strain lacking the proteinases A and B and the carboxypeptidases Y and S. Promoter trimming, MAL2-8cp stimulation and the use of a host strain deficient in four vacuolar proteinases resulted in α-glucosidase PI expression of about 13% of the soluble protein.
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  • 95
    ISSN: 0749-503X
    Keywords: TRP1 ; histone H3 ; histone H4 ; pyrophosphatase ; Kluyveromyces ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The TRP1 gene of the yeast Kluyveromyces lactis has been cloned from a genomic library by complementation of the Saccharomyces cerevisiae trpl-289 mutation. The gene was located within the clone by transposon mutagenesis and the coding region identified by DNA sequencing. This has indicated that K. lactis TRP1 encodes a 210-amino acid polypeptide which shows 53% identity to the homologous S. cerevisiae protein. The K. lactis TRP1 gene has been disrupted by substituting the S. cerevisiae URA3 gene for a large part of the TRP1 coding sequence. Replacement of the chromosomal TRP1 locus with this construction has enabled the production of non-reverting trp1- strains of K. lactis, while a genetic analysis of the disrupted allele confirmed that the TRP1 gene had been cloned. DNA sequencing has also shown that the K. lactis TRP1 sequences is flanked by genes encoding inorganic pyrophosphatase and histone H3, which we have designated IPP and HHT1 respectively. Hybridization studies have shown that in common with S. cerevisiae, K. lactis has two copies of the histone H3 gene. Each H3 gene is closely linked to a gene encoding histone H4 and in both yeast species the IPP gene is tightly linked to one of the histone gene pairs.
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  • 96
    ISSN: 0749-503X
    Keywords: Yeast ; protein ; extract ; trichloroacetic acid ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Methods currently used for the extraction of proteins from yeast involve relatively long time periods between sampling cells from a culture and analysis of their proteins by polyacrylamide gel electrophoresis-sodium dodecylsulphate. Often it is desirable to inactivate cellular metabolism rapidly after sampling and here we show that trichloroacetic acid precipitation techniques, often used for rapid extraction and inactivation of proteins from higher eukaryotes, can be adapted for use with organisms which have cell walls.
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  • 97
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    Yeast 5 (1989) 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 98
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    Yeast 5 (1989), S. ii 
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    Keywords: Life and Medical Sciences ; Genetics
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  • 99
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    Yeast 5 (1989), S. 55-72 
    ISSN: 0749-503X
    Keywords: Gene disruption ; genetic mapping ; nonsense suppression ; multibudded phenotype ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A newly isolated gene, ESS1, was shown to encode a protein required for vegetative growth in Saccharomyces cerevisiae. The nucleotide sequence of ESS1 revealed a 172 amino acid open reading frame predicting a highly basic, 19·5 kilodalton product. Although the gene was isolated by cross-hybridization with the vertebrate v-sis oncogene, the primary amino acid sequence bears only a slight resemblance to the p28sis protein. ESS1 was shown to be single copy in the yeast genome and transcriptionally active during logarithmic growth. It is located on the right arm of chromosome X, 6 centimorgans distal to ilv3. The genetic map location indicates it is not allelic to any previously characterized mutation in this organism. Both inactivation of ESS1 by gene disruption and overexpression by fusion to a heterologous promoter were detrimental to growth in both haploid and diploid cell types. Under non-permissive conditions, the terminal phenotype of strains containing a suppressible amber mutation within ESS1 was one of aberrant multibudded structures. Examination of this morphology indicates that loss of ESS1 function may lead to a defect in cytokinesis or cell separation.
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  • 100
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    Yeast 5 (1989), S. 73-77 
    ISSN: 0749-503X
    Keywords: vandate ; mitochondria ; H+ ATPase ; S. cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The effects of vandate on mitochondrial respiration and H+ ATPase activity in Saccharomyces cerevisiae were studied. A 50% inhibition of oxygen uptake in isolated mitochondria was produced by 4·4 mM-V2O5. Activity of H+ ATPase in whole mitochondria was inhibited by 50% by 5·5 μM-V2O5, in submitochondrial particles by 55 μM-V2O5; and in the chloroform-released H+ ATPase by 0·5 mM-V2O5. Vandate was also found to relieve growth inhibition caused by the mitochondrial H+ ATPase inhibitors NN′-decyclohexylcarbodiimide and oligomycin. These results imply that vanadate could affect mitochondrial respiration by interacting with the H+ ATPase in S. cerevisiae.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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