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  • 1996  (244)
  • Life Sciences (general)  (244)
  • 101
    Electronic Resource
    Electronic Resource
    Sequence analysis of a 10 kb DNA fragment from yeast chromosome VII reveals a novel member of the dnaJ family (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 145-148 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome VII ; CEG1 ; SOH1 ; DnaJ ; SCS3 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence analysis of a 10 kb DNA fragment of Saccharomyces cerevisiae chromosome VII. This sequence contains four complete open reading frames (ORFs) of greater than 100 amino acids. There are also two incomplete ORFs flanking the extremes: one of these, G2868, is the 5′ part of the SCS3 gene (Hosaka et al., 1994). ORFs G2853 and G2856 correspond to the genes CEG1, coding for the alfa subunit of the mRNA guanylyl transferase and a 3′ gene of unknown function previously sequenced (Shibagaki et al., 1992). G2864 is identical to SOH1 also reported (Fan and Klein, 1994). The translated sequence of G2861 is similar to the human dnaJ homolog. The nucleotide sequence reported here has been entered in the EMBL Data Library under the Accession Number X87252.
    Additional Material: 4 Ill.
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  • 102
    Electronic Resource
    Electronic Resource
    Structure and regulation of the Candida albicans ADH1 gene encoding an immunogenic alcohol dehydrogenase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 115-127 
    Details
    ISSN: 0749-503X
    Keywords: Candida albicans ; alcohol dehydrogenase ; gene regulation ; messenger RNA ; carbon utilization ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Candida albicans ADH1 gene encodes an alcohol dehydrogenase which is immunogenic during infections in humans. The ADH1 gene was isolated and sequenced, and the 5′- and 3′-ends of its mRNA were mapped. The gene encodes a 350 amino acid polypeptide with strong homology (70·5-85·2% identity) to alcohol dehydrogenases from Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. The cloned C. albicans ADH1 gene was shown to be functional through complementation of adh mutations and efficient production of active alcohol dehydrogenase in S. cerevisiae. Northern analysis of C. albicans RNA revealed that ADH1 mRNA levels were regulated in response to carbon source and during batch growth. During growth on glucose, ADH1 mRNA levels rose to maximum levels during late exponential growth phase and declined to low levels in stationary phase. The ADH1 mRNA was relatively abundant during growth on galactose, glycerol, pyruvate, lactate or succinate, and less abundant during growth on glucose or ethanol. Alcohol dehydrogenase levels did not correlate closely with ADH1 mRNA levels under the growth conditions studied, suggesting either that this locus is controlled at both transcriptional and post-transcriptional levels, or that other differentially regulated ADH loci exist in C. albicans.
    Additional Material: 7 Ill.
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  • 103
    Electronic Resource
    Electronic Resource
    pMPY-ZAP: A reusable polymerase chain reaction-directed gene disruption cassette for Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 129-134 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; gene disruption ; PCR-mediated disruption ; uraduction ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Gene disruption is an important method for genetic analysis in Saccharomyces cerevisiae. We have designed a polymerase chain reaction-directed gene disruption cassette that allows rapid disruption of genes in S. cerevisiae without previously cloning them. In addition, this cassette allows recycling of URA3, generating gene disruptions without the permanent loss of the ura3 marker. An indefinite number of disruptions can therefore be made in the same strain.
    Additional Material: 3 Ill.
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  • 104
    Electronic Resource
    Electronic Resource
    Independent regulation of full-length and 5′-truncated PAS5 mRNAs in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 135-143 
    Details
    ISSN: 0749-503X
    Keywords: peroxisome ; Zellweger syndrome ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Peroxisome assembly in Saccharomyces cerevisiae requires the products of several genes. In this report, the PAS5 gene has been characterized. The gene is on the left arm of chromosome X and encodes a polypeptide with similarity to the mammalian peroxisome assembly factor-1 (PAF-1). Two different length transcripts are produced from the yeast PAS5 gene. The longer mRNAs encompass an open reading frame, while the shorter transcripts initiate 46-110 base pairs downstream of the first in-frame AUG. The longer transcripts are induced four-fold on medium containing fatty acids as the sole carbon source, while the shorter transcripts are induced up to ten-fold on medium containing glycerol as a carbon source. The full-length coding sequence encodes a protein with a calculated molecular weight of 30·7 kDa. A protein of 25 kDa could be translated from the shorter transcripts and would lack a very acidic domain found in the amino-terminal extension of the longer protein. The common portion of the proteins is very basic; the calculated pI of the longer polypeptide is 9·02 and that of the shorter protein is 10·06. The PAS5 GenBank accession number is M86538.
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  • 105
    Electronic Resource
    Electronic Resource
    The sequence of 36·8 kb from the left arm of chromosome XIV reveals 24 complete open reading frames: 18 correspond to new genes, one of which encodes a protein similar to the human myotonic dystrophy kinase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 169-175 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; chromosome XIV ; Saccharomyces cerevisiae ; Myotonic Dystrophy Kinase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the complete nucleotide sequence of a 36·8 kb segment from the left arm of chromosome XIV carried by the cosmid 14-11. The sequence encodes the 5′ coding region of the PSD1 gene, the 3′ coding region of an unknown gene and 24 complete open reading frames, of which 18 correspond to new genes and six (SKO1, SCL41A, YGP1, YCK2, RPC31 and MFA2) have been sequenced previously. Of the 24 new genes, five show significant similarities to sequences present in the databanks. These include elongation factors 2 and the human myotonic dystrophy kinase. The sequence has been deposited in the EMBL databank under the Accession Number X92517.
    Additional Material: 4 Ill.
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  • 106
    Electronic Resource
    Electronic Resource
    Masthead (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320120201
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  • 107
    Electronic Resource
    Electronic Resource
    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 191-198 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320120202
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  • 108
    Electronic Resource
    Electronic Resource
    Fifteen open reading frames in a 30·8 kb region of the right arm of chromosome VI from Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 177-190 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VI ; Snf2/Rad54 helicase family ; SUP6 ; HIS2 ; CDC14 ; MET10 ; SMC2 ; QCR6 ; PHO4 ; CDC26 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of cosmid clone 9765, which contains 30·8 kb of the right arm of chromosome VI, was determined. Both strands were sequenced, with an average redundancy of 8·17 per base pair by both dye primer and dye terminator cycle sequencing methods. The G+C content of the sequence was found to be 40·3%. Fifteen open reading frames (ORFs) greater than 100 amino acids and one tRNA-Tyr gene (SUP6) were detected. Seven of the ORFs were found to encode previously identified genes (HIS2, CDC14, MET10, SMC2, QCR6, PH04 and CDC26). One ORF, 9765orfF010, was found to encode a new member of the Snf2/Rad54 helicase family. Three ORFs (9765orfR002, 9765orfR011 and 9765orfR013) were found to be homologous with Schizosaccharomyces pombe polyadenylate binding protein, Escherichia coli hypothetical 38·1-kDa protein in the BCR 5′ region, and transcription regulatory protein Swi3, respectively. The sequence may be found in the DDBJ, EMBL and GenBank nucleotide sequence databases under Accession Number D44602.
    Additional Material: 5 Ill.
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  • 109
    Electronic Resource
    Electronic Resource
    Analysis of a 36·2 kb DNA sequence including the right telomere of chromosome VI from Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 149-167 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VI ; telomere ; ORFs ; DnaJ-like protein ; mitochondrial carrier protein ; cystathionine lyase ; YMR31 ; PRE4 ; NIN1 ; HXK1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a 36·2-kb distal region containing the right telomere of chromosome VI was determined. Both strands of DNA cloned into cosmid clone 9965 and plasmid clone pEL174P2 were sequenced with an average redundancy of 7·9 per base pair, by both dye primer and dye terminator cycle sequencing methods. The G + C content of the sequence was found to be 37·9%. Eighteen open reading frames (ORFs) longer than 100 amino acids were detected. Four of these ORFs (9965orfR017, 9965orfF016, 9965orfR009 and 9965orfF003) were found to encode previously identified genes (YMR31, PRE4, NIN1 and HXK1, respectively). Six ORFs (9965orfR013, 9965orfF018, 9965orfF006, 9965orfR014, 9965orfF013 and 9965orfR020) were found to be homologous to hypothetical 121·4-kDa protein in the BCK 5′ region, Bacillus subtilis DnaJ protein, hypothetical Trp-Asp repeats containing protein in DBP3-MRPL27, putative mitochondrial carrier YBR291C protein, Salmonella typhimurium nicotinate-nucleotide pyrophosphorylase, and Escherichia coli cystathionine β-lyase, respectively. The putative proteins encoded by 9965orfF018, 9965orfR014 and 9965orfR020 were found to be, respectively, a new member of the family of DnaJ-like proteins, the mitochondrial carrier protein and cystathionine lyase. The nucleotide sequence reported here has been deposited in the DDBJ/GenBank/EMBL data library under Accession Number D44597.
    Additional Material: 7 Ill.
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  • 110
    Electronic Resource
    Electronic Resource
    Selection of yeast cells with a higher plasmid copy number in a Saccharomyces cerevisiae autoselection system (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 199-205 
    Details
    ISSN: 0749-503X
    Keywords: S. cerevisiae ; autoselection plasmid ; flow cytometry ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Autoselection systems allow the selection of a genetically engineered population independently of the growth medium composition. The structure of a Saccharomyces cerevisiae population transformed with an autoselection plasmid, in which a carbon-source-dependent modulation of the plasmid copy number occurs, was analysed.By means of flow cytometric procedures we tested the cell viability, dynamics of growth and heterologous protein production at single cell level. Such analyses allow the identification and the tracking of a specific cellular sub-population with a higher plasmid copy number which arises after the carbon source shift. The effects of the cellular plasmid distribution on the dynamics of growth are also discussed.
    Additional Material: 5 Ill.
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  • 111
    Electronic Resource
    Electronic Resource
    Identification of ASK10 as a multicopy activator of Skn7p-dependent transcription of a HIS3 reporter gene (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 267-272 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; ASK10 ; SKN7 ; two-component ; response regulator ; transcription factor ; U27209 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Recent evidence has demonstrated that the yeast Skn7p appears to act as a ‘response regulator’ in a eukaryotic ‘two-component’ signal transduction pathway. A search to identify possible regulators of the SKN7 mediated ‘two- component’ regulatory system has identified Ask10p as a novel potential transcription factor. The ASK10 sequence has been deposited in GenBank with Accession Number U27209.
    Additional Material: 3 Ill.
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  • 112
    Electronic Resource
    Electronic Resource
    A rapid and selective assay for measuring cell surface hydrophobicity of brewer's yeast cells (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 207-213 
    Details
    ISSN: 0749-503X
    Keywords: brewer's yeast ; cell surface ; flocculation ; hydrophobicity ; magnobeads ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A rapid and selective assay was developed to measure cell surface hydrophobicity of brewer's yeast cells. During this so-called magnobead assay, bottom-fermenting yeast cells adhere to paramagnetic, polystyrene-coated latex beads which can easily be removed from the cell suspension by using a (samarium-cobalt) magnet. At pH 4·5, electrostatic repulsion between yeast cells and latex beads was found to be minimal and yeast cell adhesion was predominantly based on hydrophobic interactions. The percentage of cells adhering to the beads could be calculated and provided a measure for cell surface hydrophobicity.Cell surface hydrophobicity measured by the magnobead assay was found to yield similar results, as did determination of contact angles of water droplets on a layer of yeast cells, a standard method for measuring surface hydrophobicity. However, the magnobead assay has the following advantages: (i) it is a quick and simple method, and, more significantly, (ii) hydrophobicity can be measured under physiological conditions. Use of the magnobead assay confirmed that a higher level of cell surface hydrophobicity is correlated with stronger flocculence of brewer's lager yeast cells.
    Additional Material: 4 Ill.
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  • 113
    Electronic Resource
    Electronic Resource
    Sequencing of a 17·6 kb segment on the right arm of yeast chromosome VII reveals 12 ORFs, including CCT, ADE3 and TR-I genes, homologues of the yeast PMT and EF1G genes, of the human and bacterial electron-transferring flavoproteins (β-chain) and of the Escherichia coli phosphoserine phosphohydrolase, and five new ORFs (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 273-280 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome sequencing ; chromosomeVII ; CCT ; ADE3 ; TR-1 ; PMT ; EF19 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A 17·6 kb DNA fragment from the right arm of chromosome VII of Saccharomyces cerevisiae has been sequenced and analysed. The sequence contains twelve open reading frames (ORFs) longer than 100 amino acids. Three genes had already been cloned and sequenced: CCT, ADE3 and TR-I. Two ORFs are similar to other yeast genes: G7722 with the YAL023 (PMT2) and PMT1 genes, encoding two integral membrane proteins, and G7727 with the first half of the genes encoding elongation factors 1γ, TEF3 and TEF4. Two other ORFs, G7742 and G7744, are most probably yeast orthologues of the human and Paracoccus denitrificans electron-transferring flavoproteins (β chain) and of the Escherichia coli phosphoserine phosphohydrolase. The five remaining identified ORFs do not show detectable homology with other protein sequences deposited in data banks. The sequence has been deposited in the EMBL data library under Accession Number Z49133.
    Additional Material: 6 Ill.
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  • 114
    Electronic Resource
    Electronic Resource
    PCR-synthesis of marker cassettes with long flanking homology regions for gene disruptions in S. cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 259-265 
    Details
    ISSN: 0749-503X
    Keywords: Yeast genome analysis ; Functional analysis ; G418 resistance ; PCR-based gene disruption ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A PCR-method for fast production of disruption cassettes is introduced, that allows the addition of long flanking homology regions of several hundred base pairs (LFH-PCR) to a marker module. Such a disruption cassette was made by linking two PCR fragments produced from genomic DNA to kanMX6, a modification of dominant resistance marker making S. cerevisiae resistant to geneticin (G418). In a first step, two several hundred base pairs long DNA fragments from the 5′- and 3′-region of a S. cerevisiae gene were amplified in such a way that 26 base pairs extensions homologous to the kanMX6 marker were added to one of their end. In a second step, one strand of each of these molecules then served as a long primer in a PCR using kanMX6 as template. When such a LFH-PCR-generated disruption cassette was used instead of a PCR-made disruption cassette flanked by short homology regions, transformation efficiencies were increased by at least a factor of thirty. This modification will therefore also help to apply PCR-mediated gene manipulations to strains with decreased transformability and/or unpredictable sequence deviations.
    Additional Material: 1 Ill.
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  • 115
    Electronic Resource
    Electronic Resource
    Saccharomycodes ludwigii has seven chromosomes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 237-240 
    Details
    ISSN: 0749-503X
    Keywords: electrophoretic karyotype ; linkage group ; Saccharomycodes ludwigii ; tetrad analysis ; yeast ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Tetrad distributions for 108 different gene pairs in 1346 asci of 113 diploids heterozygous for various combinations of 24 genes in Saccharomycodes ludwigii were investigated. Tetratype tetrads occurred only rarely and the 24 genes tested were classified into seven linkage groups. Electrophoretic karyotypes of three independent strains of S'codes ludwigii showed seven bands of chromosome-sized DNA having molecular sizes of 0·8 to 2·3 Mb with strain-specific polymorphic chromosomal DNAs as determined based on their migration distances.
    Additional Material: 2 Ill.
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  • 116
    Electronic Resource
    Electronic Resource
    Sequencing and analysis of 51 kb on the right arm of chromosome XV from Saccharomyces cerevisiae reveals 30 open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 281-288 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; DNA sequencing project ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a region of 51 kb of the right arm from chromosome XV of Saccharomyces cerevisiae. The sequence contains 30 open reading frames (ORFs) of more than 100 amino acid residues. Thirteen new genes have been identified. Thirteen ORFs correspond to known yeast genes. One delta element and one tRNA gene were identified. Upstream of the RPO31 gene, encoding the largest subunit of RNA polymerase III, lies a Abf1p binding site. The nucleotide sequence data reported in this paper are available in the EMBL, GenBank and DDBJ nucleotide sequence databases under the Accession Number X90518.
    Additional Material: 2 Ill.
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  • 117
    Electronic Resource
    Electronic Resource
    Isolation and characterization of the Schizosaccharomyces pombe gene encoding transcript elongation factor TFIIS (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 227-236 
    Details
    ISSN: 0749-503X
    Keywords: transcription elongation ; TFIIS ; SII ; 6-azauracil ; Schizosaccharomyces pombe ; fission yeast ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A gene designated tfs1 has been isolated from Schizosaccharomyces pombe based on its similarity to genes encoding transcription elongation factor TFIIS. The nucleotide sequence of the tfs1 gene predicts a polypeptide with similarity to mammalian, Drosophila and Saccharomyces cerevisiae TFIIS. A haploid Sz. pombe strain with tfs1 deleted from the genome is viable. Thus, tfs1 is not essential for viability. However, deletion of tfs1 results in slow growth and increased sensitivity to the drug 6-azauracil, a phenotype similar to that of a S. cerevisiae strain deleted for the gene encoding TFIIS. The DNA sequence of tfs1 has been deposited in GenBank under Accession Number U20526.
    Additional Material: 6 Ill.
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  • 118
    Electronic Resource
    Electronic Resource
    Malolactic fermentation by engineered Saccharomyces cerevisiae as compared with engineered Schizosaccharomyces pombe (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 215-225 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Lactococcus lactis ; malolactic enzyme ; malolactic fermentation ; heterologous expression ; NMR ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ability of yeast strains to perform both alcoholic and malolactic fermentation in winemaking was studied with a view to achieving a better control of malolactic fermentation in enology. The malolactic gene of Lactococcus lactis (mleS) was expressed in Saccharomyces cerevisiae and Schizosaccharomyces pombe. The heterologous protein is expressed at a high level in cell extracts of a S. cerevisiae strain expressing the gene mleS under the control of the alcohol dehydrogenase (ADH1) promoter on a multicopy plasmid. Malolactic enzyme specific activity is three times higher than in L. lactis extracts. Saccharomyces cerevisiae expressing the malolactic enzyme produces significant amounts of l-lactate during fermentation on glucose-rich medium in the presence of malic acid. Isotopic filiation was used to demonstrate that 75% of the l-lactate produced originates from endogenous l-malate and 25% from exogenous l-malate. Moreover, although a small amount of exogenous l-malate was degraded by S. cerevisiae transformed or not by mleS, all the exogenous degraded l-malate was converted into l-lactate via a malolactic reaction in the recombinant strain, providing evidence for very efficient competition of malolactic enzyme with the endogenous malic acid pathways. These results indicate that the sole limiting step for S. cerevisiae in achieving malolactic fermentation is in malate transport. This was confirmed using a different model, S. pombe, which efficiently degrades l-malate. Total malolactic fermentation was obtained in this strain, with most of the l-malate converted into l-lactate and CO2. Moreover, l-malate was used preferentially by the malolactic enzyme in this strain also.
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  • 119
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    Electronic Resource
    Corrigendum to: The sequence of a 44 458 bp fragment located on the left arm of chromosome XIV from Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 297-297 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 120
    Electronic Resource
    Electronic Resource
    The terminal protein of the linear DNA plasmid pGKL2 shares an N-terminal domain of the plasmid-encoded DNA polymerase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 241-246 
    Details
    ISSN: 0749-503X
    Keywords: terminal protein ; pGKL2 plasmid ; DNA polymerase ; Kluyveromyces lactis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The 36K protein attached at the 5′ end of the linear DNA plasmid pGKL2 from the yeast Kluyveromyces lactis was first purified and characterized. The terminal protein was purified from cells (1 kg wet weight) by ammonium sulphate precipitation and two rounds of centrifugation to equilibrium in CsCl gradients. The pGKL2 was present only in the post-microsomal supernatant. Approximately 10 mg of the purified pGKL2 was recovered and digested with DNase I. The terminal protein (final ca. 0·8 mg) was homogeneous by electrophoresis and we determined the N-terminal amino acid sequence up to ten residues, showing that it existed in the cryptic N-terminal domain of pGKL2-ORF2 (DNA polymerase) sequence.
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  • 121
    Electronic Resource
    Electronic Resource
    Sequencing of a 9·2 kb telomeric fragment from the right arm of Saccharomyces cerevisiae chromosome XIV (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 289-295 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome XIV ; right telomere ; sub-telomeric repeats ; mannitol dehydrogenase homolog ; YCR007/YKL219-like ; PAU1- like ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the complete sequence of a 9·2 kb fragment next to and including the right telomere of Saccharomyces cerevisiae chromosome XIV. Four open reading frames (ORFs) longer than 100 amino acids were observed in the sequenced segment. One ORF (378 codons) does not show any significant homology with proteins in the databases and corresponds to a putative new gene. Two ORFs are almost identical to the known YCR007/YKL219 and PAU1-like hypothetical protein families already identified on several S. cerevisiae chromosomes. These ORFs, whose function is unknown, are generally associated with sub-telomeric regions of chromosomes. The fourth one shows significant identities with bacterial mannitol dehydrogenases. It could be a yeast gene implicated in the metabolism of mannitol (or a related substrate). The sequence has been deposited in the EMBL data library under Accession Number X86790.
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  • 122
    Electronic Resource
    Electronic Resource
    Discrimination between fortuitous and biologically constrained open reading frames in DNA sequences of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 369-384 
    Details
    ISSN: 0749-503X
    Keywords: open reading frames ; random DNA sequence ; functional analysis ; Saccharomyces cerevisiae ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The systematic sequencing of the yeast genome has raised the problem of the biological significance of the open reading frames (ORFs) revealed: it is possible that some of these are fortuitous. To avoid the analysis of such fortuitous ORFs, a minimum length of 100 sense codons was adopted. Nevertheless, the presence of fortuitous ORFs of more than 100 codons cannot be excluded. Thus, in the context of functional analysis, a method for discrimination between fortuitous and biologically active ORFs may be useful. The discrimination method described here is based on multiple criteria: ORF length, codon bias, and both amino-acid and dipeptide composition of the corresponding polypeptide. The thresholds for each criterion are based on the comparison between two learning sets: one drawn from random DNA sequences and the second from known genes. The method was validated by two test sets (one random and one biological) and then applied to the ORFs of chromosomes I, II, III, V, VIII, IX and XI. This method predicts 123 fortuitous ORFs among the 1773 identified on these chromosomes.
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    The sequence of a 21·3kb DNA fragment from the left arm of yeast chromosome XIV reveals LEU4, MET4, POL1, RAS2, and six new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 403-409 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XIV ; LEU4 ; MET4 ; POL1 ; RAS2 ; inositol-1,4,5-trisphosphate 5-phosphatase ; Lowe's syndrome ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a fragment of 21 308 bp from the left arm of Saccharomyces cerevisiae chromosome XIV has been determined. Analysis of the sequence revealed 13 open reading frames (ORFs) longer than 300 bp, four of which correspond to the previously identified genes LEU4, MET4, POL1 and RAS2. One putative protein, N2160, shares considerable homology (32% identity) with a hypothetical protein encoded by a gene located on chromosome XV as well as with human OCRL protein (36% identity), involved in Lowe's syndrome. N2185 contains ten predicted transmembrane segments and is similar to another putative protein (YKL146) from yeast. The sequence of the reported DNA fragment has been submitted to the EMBL data library under Accession Number Z50161.
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    Flocculation of the yeast Candida famata (Debaryomyces hansenii): An essential role for peptone (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 415-423 
    Details
    ISSN: 0749-503X
    Keywords: flocculation ; Candida famata ; peptone ; lectin ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Aggregation of Candida famata (Debaryomyces hansenii) is consistent with being a form of lectin-mediated yeast flocculation. Flocculation of C. famata is unusual in that it requires the presence of peptone, either in the growth medium or added later to harvested cells in buffer. Flocculation after peptone addition was rapid, being largely complete within 10 min. Heat-killed cells also flocculated, arguing for direct participation of peptone in the flocculation binding mechanism.Flocculent C. famata cells progressively lost the ability to flocculate when washed with EDTA. Flocculation was fully restored by peptone addition; calcium addition was without effect. C. famata cells were able to agglutinate erythrocytes in the presence or absence of peptone. Pronase E-treated yeast lost both the ability to haemagglutinate and self-flocculate. Haemagglutination was not diminished by progressive EDTA washing, suggesting that surface lectins remained present and active on the yeast cell walls. Non-flocculating C. famata cells mutually flocculated with non-flocculent Schizosaccharomyces pombe cells, shown to have surface-exposed galactose residues. Mutual flocculation was lost following treatment of C. famata with Pronase E.It was concluded that the cell wall of C. famata contains lectins enabling haemagglutination and mutual flocculation but lacks carbohydrate receptors for these lectins. This yeast self-flocculates only via bridging multi-valent carbohydrates; these being present in peptone.
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    Sequence of ptb1, a Gene for the β subunit of the type-II geranylgeranyltransferase from the fission yeast Schizosaccharomyces pombe (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 479-483 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated and sequenced the ptb1 gene from the fission yeast Schizosaccharomyces pombe. Sequence analysis suggests that Ptb1 is the β subunit of the type-II geranylgeranyltransferase that is responsible for geranylgeranylation of the Rab-like YPT proteins in this yeast. The sequence has been deposited in the EMBL data library under the Accession Number X92183.
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    Secretion of a variant of human single-chain urokinase-type plasminogen activator without an N-glycosylation site in the methylotrophic yeast, Pichia pastoris and characterization of the secreted product (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 541-553 
    Details
    ISSN: 0749-503X
    Keywords: Pichia pastoris ; single-chain urokinase-type plasminogen activator ; Mucor rennin prepeptide ; proteolysis ; secretion ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Human single-chain urokinase-type plasminogen activator without an N-glycosylation site (scu-PA-Q302) was produced in the methylotrophic yeast, Pichia pastoris using the shortened prepeptide sequence of a fungal aspartic proteinase, Mucor pusillus rennin (MPR). The level of urokinase-type plasminogen activator (u-PA) immunoreactive material in YPM medium was 0·47 mg/l; however, most of the secreted product had been processed to smaller polypeptides. The N-terminal amino acid sequence of major species was identical to that of the low molecular weight two-chain u-PA. Some approaches to minimizing the proteolysis of scu-PA-Q302 were attempted. Addition of Triton X-100, l-arginine and ammonium phosphate to the YPM medium minimized the proteolysis of scu-PA-Q302 and increased the yield of immunoreactive material to approximately 5 mg/l. Use of proteinase A- or proteinase B-deficient strains of yeast did not reduce the degradation. Co-expression of scu-PA-Q302 and urinary trypsin inhibitor resulted in partial reduction of the major species of proteolysis.Scu-PA-Q302 was purified from the culture supernatant of the improved medium by two successive chromatographies on Phenyl-Sepharose and S-Sepharose. The purified protein had a molecular weight of 47 kDa. It did not contain detectable N-linked oligosaccharides, but contained O-linked oligosaccharides attached to the light chain. N-terminal amino acid sequencing of the purified preparation showed that the shortened prepeptide sequence of MPR was correctly processed by the Pichia yeast. Scu-PA-Q302 closely resembles natural scu-PA with respect to its enzymatic activity against the chromogenic substrate S-2444 and its in vitro fibrinolytic properties.
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    Subtelomeric duplications in Saccharomyces cerevisiae chromosomes III and XI: Topology, arrangements, corrections of sequence and strain-specific polymorphism (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 583-591 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome III ; chromosome XI ; sequence ; duplication ; translocation ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the sequence of a 3·42 kb segment from the left arm of chromosome III (coordinates 5394-8815 of Oliver et al., 1992). Instead of four open reading frames (ORFs) listed previously, the verified sequence reveals the presence of only one ORF, renamed YCL070/73c, encoding a protein of 615 amino acids. The putative product of ORF YCL070/73c shows 98·5% identity and 99% similarity with the protein of the same length encoded by ORF YKR106w from the right arm of chromosome XI and displays a topology characteristic for the Major Facilitators Superfamily of membrane proteins. These corrections will be deposited in the EMBL data library under the Accession Number X59720. In strain S288C the subtelomeric sequence 4319-11 215 of chromosome III is 98·3% identical with the subtelomeric sequence of 658 204-665 061 from the right arm of chromosome XI. Using various subtelomeric probes from chromosome III (coordinates 2097-3646 of S288C) we have analysed eight different Saccharomyces cerevisiae strains and the closely related species S. douglasii: some S. cerevisiae strains have additional duplications and longer chromosomes XI; in all strains chromosome III contains the 1200-11 000 segment (strain FL100 is disomic) while S. douglasii does not show any hybridization in this region.
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    Sustained oscillations in free-energy state and hexose phosphates in yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 731-740 
    Details
    ISSN: 0749-503X
    Keywords: oscillation ; glycolysis ; yeast ; Saccharomyces cerevisiae ; intracellular metabolites ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In a population of intact cells of the yeast Saccharomyces cerevisiae the dynamics of glycolytic metabolism were investigated under the condition of sustained oscillations. At 5-s intervals cells were quenched in -40°C methanol, extracted and the intracellular concentrations of glycolytic metabolites, adenine nucleotides and phosphate were analysed. Oscillations were found for the glycolytic intermediates glucose 6-phosphate, fructose 6-phosphate and fructose 1,6-bisphosphate. At variance with earlier reports on transient glycolytic oscillations, some intermediates further down the glycolytic pathway did not oscillate significantly, even though NADH did. In addition, the adenylate energy charge and the free energy of ATP hydrolysis oscillated significantly. Dynamic coupling through the latter may be responsible for this effective compartmentation of glycolytic dynamics.
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    Sequence analysis of a 40·7 kb segment from the left arm of yeast chromosome X reveals 14 known genes and 13 new open reading frames including homologues of genes clustered on the right arm of chromosome XI (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 787-797 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome X ; VPS35 ; INO1 ; SnR128 ; SnR190 ; MP12 ; YAK1 ; RPB4 ; YUR1 ; TIF2 ; MRS3 ; URA2 ; RSP25 homologue ; leucine zippers ; membrane proteins ; glycogenin glycosyltransferase ; human phospholipase D ; chromosome XI ; gene cluster translocation/recombination ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete nucleotide sequence of a 40·7 kb segment about 130 kb from the left end of chromosome X of Saccharomyces cerevisiae was determined from two overlapping cosmids. Computer analysis of that sequence revealed the presence of the previously known genes VPS35, INO1, SnR128, SnR190, MP12, YAK1, RPB4, YUR1, TIF2, MRS3 and URA2, three previously sequenced open reading frames (ORFs) of unknown function 5′ of the INO1, 5′ of the MP12 and 3′ of the URA2 genes and 13 newly identified ORFs. One of the new ORFs is homologous to mammalian glycogenin glycosyltransferases and another has similarities to the human phospholipase D. Some others contain potential transmembrane regions or leucine zipper motifs. The existence of yeast expressed sequence tags for some of the newly identified ORFs indicates that they are transcribed. A cluster of six genes within 10 kb (YUR1, TIF2, two new ORFs, an RSP25 homologue and MRS3) have homologues arranged similarly within 28·5 kb on the right arm of chromosome XI. The sequence has been deposited in the EMBL data library under Accession Number X87371.
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    Lambda clone B22 contains a 7676 bp genomic fragment of Saccharomyces cerevisiae chromosome VII spanning the VAM7-SPM2 intergenic region and containing three novel transcribed open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 799-807 
    Details
    ISSN: 0749-503X
    Keywords: ypt/rab-like proteins ; GTPases ; zinc-finger proteins ; duplications ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A genomic clone of 7676 bp designated B22 from Saccharomyces cerevisiae has been sequenced. The 5′ end matches the previously described gene, VAM7, and the 3′ end matches the previously described gene, SPM2, both of which have been assigned to the left arm of chromosome VII. The intergenic region contains three transcribed open reading frames (ORFs). The first is related to an uncharacterized ORF of Bacillus subtilis and more weakly to MesJ in Escherichia coli; this is found as a single transcript of 1·1 kb by Northern blotting. The second ORF encodes a small ras-like GTPase of 222 residues with strong homology to yeast Ypt8p and to mammalian Rab11; this is found as a single transcript of 1·1 kb by Northern blotting. The third ORF generates a transcript of 1·6 kb and encodes a protein of 382 residues including a perfect match to the consensus sequence of a C2H2 zinc finger domain; it shares a strong homology with yeast Mig1p and Cre-A from Aspergillus, Emericella and E. coli. This ORF also has a striking similarity to a putative 43 kDa zinc finger protein encoded by an ORF (YEL8) immediately downstream of YPT8, raising the possibility that a region between VAM7 and SPM2 on chromosome VII arose as a duplication of the YPT8-YEL8 region of chromosome V, followed by a translocation. The sequence of the clone described has been deposited with the GenBank database under Accession Number U33754.
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    The 2μm plasmid of laboratory yeast strains is a type-1/type-2 hybrid (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 809-813 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; 2μm plasmid ; molecular evolution ; homeologous recombination ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Industrial yeast strains carry one of two homeologous 2μm plasmids designated as type-1 or type-2. The 2μm plasmid, Scp1, found in common laboratory strains of Saccharomyces cerevisiae is considered a type-2 plasmid, since the ori, STB, RAF and REP1 loci and intergenic sequences of the right-unique region of Scp1 are homologous to the corresponding loci in industrial strain type-2 plasmids. However, within both its 599 bp inverted repeats Scp1 has 142-bp sequences homologous to the bakers' yeast type-1 plasmid. DNA sequence analyses and oligonucleotide hybridizations indicate that the 142-bp insertion in Scp1 was probably due to homeologous recombination between type-1 and type-2 plasmids. These results suggest that some of the plasmid and chromosomal sequence polymorphisms seen in laboratory yeast strains result from homeologous recombination in their ancestral breeding stock.
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    Masthead (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320120801
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    Flavin adenine dinucleotide binding is the crucial step in alcohol oxidase assembly in the yeast Hansenula polymorpha (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 917-923 
    Details
    ISSN: 0749-503X
    Keywords: alcohol oxidase ; flavin adenine dinucleotide ; peroxisomes ; Hansenula polymorpha ; protein translocation ; protein assembly ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied the role of flavin adenine dinucleotide (FAD) in the in vivo assembly of peroxisomal alcohol oxidase (AO) in the yeast Hansenula polymorpha. In previous studies, using a riboflavin (Rf) autotrophic mutant, an unequivocal judgement could not be made, since Rf-limitation led to a partial block of AO import in this mutant. This resulted in the accumulation of AO precursors in the cytosol where they remained separated from the putative peroxisomal AO assembly factors. In order to circumvent the peroxisomal membrane barrier, we have now studied AO assembly in a peroxisome-deficient/Rf-autotrophic double mutant (Δper1.rif1) of H. polymorpha. By sucrose density centrifugation and native gel electrophoresis, three conformations of AO were detected in crude extracts of Δper1.rif1 cells grown under Rf-limitation, namely active octameric AO and two inactive, monomeric forms. One of the latter forms lacked FAD; this form was barely detectable in extracts wild-type and Δper1 cells, but had accumulated in the cytosol of rif1 cells. The second form of monomeric AO contained FAD; this form was also present in Δper1 cells but absent/very low in wild-type and rif1 cells. In vivo only these FAD-containing monomers associate into the active, octameric protein. We conclude that in H. polymorpha FAD binding to the AO monomer is mediated by a yet unknown peroxisomal factor and represents the crucial and essential step to enable AO oligomerization; the actual octamerization and the eventual crystallization in peroxisomes most probably occurs spontaneously.
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    Cloning, mapping and characterization of a genomic copy of the Lipomyces kononenkoae α-amylase-encoding gene (LKA1) (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 925-937 
    Details
    ISSN: 0749-503X
    Keywords: α-amylase ; LKA1 ; Lipomyces kononenkoae ; Saccharomyces cerevisiae ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The expression in Saccharomyces cerevisiae and Schizosaccharomyces pombe of a cDNA copy of the Lipomyces kononenkoae IGC4052B α-amylase gene (LKA1), linked to the phosphoglycerate kinase gene (PGK1) promoter, resulted in the extracellular production of biologically active α-amylase (LKA1). However, transformation of S. cerevisiae and Schiz. pombe with a cosmid clone containing the complete genomic copy of LKA1, expressed from its native promoter, did not result in secretion of active α-amylase by any of the transformants. When the cDNA copy of LKA1 was expressed in S. cerevisiae under control of the wild-type L. kononenkoae promoter, biologically active α-amylase was secreted into the culture medium, indicating the recognition of the LKA1 promoter in S. cerevisiae. Sequence analysis of the GC-rich LKA1 promoter revealed canonical sequences that are homologous to the TATAAA, CAAT and CCAAT boxes and GCN4-binding sites that are present in several promoter sequences of S. cerevisiae. Primer extension analysis of LKA1 transcripts in L. kononenkoae indicated major initiation sites at nucleotides -64 and -65. S. cerevisiae and Schiz. pombe cells transformed with a plasmid containing the open reading frame of the genomic copy of LKA1, linked to the PGK1 promoter, did not produce α-amylase. Polymerase chain reaction mapping and sequence analysis revealed the presence of a 61-bp intron in the genomic copy of LKA1 that impaired synthesis of biologically active α-amylase in S. cerevisiae and Schiz. pombe. This intron contains donor, acceptor and branch sequences that correlate with the consensus sequences identified in the introns of split genes from Schiz. pombe and mammals. Pulsed-field gradient gel electrophoresis resolved at least eight chromosomal DNAs for L. kononenkoae IGC4052B and chromoblot analysis indicated that LKA1 is located on the second smallest chromosome, designated chromosome II.
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    The translation initiation factor eIF4A from Schizosaccharomyces pombe is closely related to its mammalian counterpart (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 977-981 
    Details
    ISSN: 0749-503X
    Keywords: DEAD-box family ; translation ; yeast ; sequence homology ; fission yeast ; RNA helicase ; cDNA ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated a cDNA clone encoding eIF4A from Schizosaccharomyces pombe. The deduced protein sequence is similar in length and sequence to other eIF4A proteins and exhibits highest similarity with the mammalian eIF4A protein. Hybridization with genomic DNA reveals two eIF4A genes located on two different chromosomes. This sequence has been deposited in the EMBL Data Library under Accession Number X80796.
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    Masthead (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320121001
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    Identification of two CyP-40-like cyclophilins in Saccharomyces cerevisiae, one of which is required for normal growth (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 943-952 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; cyclophilin ; transcriptional regulator ; budding yeast ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the analysis of two Saccharomyces cerevisiae cyclophilins, Cpr6 and Cpr7, identified by their ability to interact in vivo with the transcriptional regulator Rpd3. Both cyclophilins have an extended carboxy-terminal region containing a three-unit tetratricopeptide repeat (TPR) motif and share significant amino acid identity with the mammalian cyclophilin CyP-40. Neither CPR6 nor CPR7 is essential but deletion of CPR7 results in a significant impairment of the rate of cell division. This is the first demonstration that a member of the cyclophilin family is required for normal cell growth. The nucleotide sequences encoding CPR6 and CPR7 have been deposited in GenBank (U48867 and U48868).
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    Isolation of a gene encoding a G protein α subunit involved in the regulation of cAMP levels in the yeast Kluyveromyces lactis (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1125-1133 
    Details
    ISSN: 0749-503X
    Keywords: G proteins ; α subunit ; signal transduction ; Kluyveromyces lactis ; cAMP ; yeast ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using chromosomal DNA from Kluyveromyces lactis as template and oligodeoxynucleotides designed from conserved regions of various G protein alpha subunits we were able to amplify by the polymerase chain reaction two products of approximately 0·5 kb (P-1) and 0·8 kb (P-2). Sequencing showed that these two fragments share high homology with genes coding for the Gα subunits from different sources. Using the P-1 fragment as a probe we screened a genomic library from K. lactis and we cloned a gene (KlGPA2) whose deduced amino acid sequence showed, depending on the exact alignment, 62% similarity and 38% identity with Gpa1p and 76% similarity and 63% identity with Gpa2p, the G protein α subunits from Saccharomyces cerevisiae. KlGPA2 is a single-copy gene and its disruption rendered viable cells with significantly reduced cAMP level, indicating that this Gα subunit may be involved in regulating the adenylyl cyclase activity, rather than participating in the mating pheromone response pathway. KlGpa2p shares some structural similarities with members of the mammalian Gαs family (stimulatory of adenylyl cyclase) including the absence in its N-terminus of a myristoyl-modification sequence. The sequence reported in this paper has been deposited in the GenBank data base (Accession No. L45105).
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    Regulation of sulphate assimilation in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1153-1162 
    Details
    ISSN: 0749-503X
    Keywords: sulphate assimilation ; cysteine regulon ; O-acetylserine ; serineO-acetyltransferase ; Saccharomyces cerevisiae ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We examined how the activity of O-acetylserine and O-acetylhomoserine sulphydrylase (OAS/OAH) SHLase of Saccharomyces cerevisiae is affected by sulphur source added to the growth medium and genetic background of the strain. In a wild-type strain, the activity was repressed if methionine, cysteine or glutathione was added to the growth medium. However, in a strain deficient of cystathionine γ-lyase, cysteine and glutathione were repressive, but methionine was not. In strains deficient of serine O-acetyltransferase (SATase), OAS/OAH SHLase activity was low regardless of sulphur source and was further lowered by cysteine and glutathione, but not by methionine. From these observations, we concluded that S-adenosylmethionine should be excluded from being the effector for regulation of OAS/OAH SHLase. Instead, we suspected that S. cerevisiae would have the same regulatory system as Escherichia coli for sulphate assimilation; i.e. cysteine inhibits SATase to lower the cellular concentration of OAS which is required for induction of the sulphate assimilation enzymes including OAS/OAH SHLase. Subsequently, we obtained data supporting this speculation.
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    Identification of a group-i intron within the 25S rDNA from the yeast Arxula adeninivorans (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1201-1208 
    Details
    ISSN: 0749-503X
    Keywords: Arxula adeninivorans ; 25S rDNA ; intron ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The 25S rDNA of the yeast Arxula adeninivorans LS3 has been cloned from a genomic library and sequenced. This DNA could be localized on chromosome 1 from A. adeninivorans and comprised 3790bp. The DNA sequence from this rDNA of the strain LS3 is very similar to the 25S rDNA of Candida albicans (91·7%), Saccharomyces cerevisiae (90·5%), Schizosaccharomyces pombe (83·8%) and Mucor racemosus (79·2%).Additionally a 411bp insertion could be localized within the 25S rDNA. This intervening sequence, which is devoid of any long open reading frame, is a group-IC intron as revealed from its site of insertion, predicted secondary structure, and its self-splicing capability. The Arxula intron is intermediate in structure and sequence between the ribosomal introns of Tetrahymena thermophila and C. albicans. The nucleotide sequence reported in this paper has been entered in the GenBank/EMBL data libraries and assigned Accession Number Z50840.
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    Masthead (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320121201
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    Reconstitution of lactate proton symport activity in plasma membrane vesicles from the yeast Candida utilis (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1263-1272 
    Details
    ISSN: 0749-503X
    Keywords: membrane vesicles ; lactic acid transport ; Candida utilis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Lactic acid transport was studied in plasma membrane vesicles from the yeast Candida utilis IGC 3092 which were fused with liposomes containing cytochrome c oxidase. After the addition of an electron donor system, these hybrid membrane vesicles were able to generate a proton-motive force of about -150mV, inside alkaline and negative. In vesicles prepared from lactic acid-grown cells, the uptake of labelled lactic acid, at pH 6·2, under energized conditions, was expressed by a kinetics consistent with the involvement of a mediated transport system. This carrier exhibited a substrate specificity pattern identical to the one found for the lactate-proton symport in intact cells. The transport of labelled lactic acid was accumulative and strongly sensitive to the effects of the protonophore carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone, consistent with the involvement of the proton-motive force in acid uptake, hence with the presence of a proton symport for lactate. Dissipation of the transmembrane electric potential by valinomycin did not have a significant effect on lactate accumulation, whereas abolishing the transmembrane pH gradient (ΔpH) by nigericin prevented the accumulation and led to a rapid efflux of the accumulated acid. The data support that the ΔpH is the main component of the proton-motive force involved in the transport of the acid and its accumulation. The lactate-proton symport stoichiometry was 1:1, being independent of the pH. Vesicles prepared from glucose-grown cells did not display the capacity to transport and accumulate lactate. However, activity for the carrier was also reconstituted in vesicles obtained from glucose-grown cells after incubation in buffer containing lactic acid. These results were consistent with those obtained in intact cells, which demonstrated that the lactate-proton symport of the yeast C. utilis is inducible.
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  • 143
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    Genetic aspects of carbon catabolite repression of the STA2 glucoamylase gene in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1297-1300 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; extracellular glucoamylase ; carbon catabolite repression ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Three regulatory genes, known to be required for glucose repression/derepression of some genes in Saccharomyces cerevisiae, were disrupted to study their effects on the carbon-source regulation of the STA2 glucoamylase gene expression. Using a STA2-lacZ fusion it was found that: (1) the MIG1 gene is dispensable for the repression of the STA2 gene; (2) there are two components in the carbon-source repression of STA2: HXK2-dependent and HXK2-independent; and (3) the HAP2 gene seems to be involved in repression rather than activation of the STA2 expression.
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  • 144
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    Masthead (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320121301
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    The sequence of a 16691bp segment of Saccharomyces cerevisiae chromosome IV identifies the DUN1, PMT1, PMT5, SRP14 and DPR1 genes, and five new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1377-1384 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome IV ; DUN1 ; PMT1 ; PMT5 ; SRP14 ; DPR1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: As part of the European BIOTECH programme, the nucleotide sequence of a 16691bp fragment from the left arm of chromosome IV of Saccharomyces cerevisiae has been deduced. Analysis of the sequence reveals the presence of 13 open reading frames (ORFs) larger than 100 codons. Five of these were previously identified as genes DUN1, PMT1, PMT5, SRP14 and DPR1. One putative protein, D2371p, contains an ATP-GTP binding site, and shares homology to the ArsA component of an Escherichia coli arsenical pump. No significant homology to any known protein has been found for the other ORFs. D2378p contains a zinc finger domain. The nucleotide sequence has been deposited at EMBL, with Accession Number X95644.
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  • 146
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    Sequence and analysis of an aldose (xylose) reductase gene from the xylose-fermenting yeast Pachysolen tannophilusThe mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned. (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1367-1375 
    Details
    ISSN: 0749-503X
    Keywords: pentose phosphate pathway ; ethanol ; fermentation ; aldehyde reductase ; ρ-crystallin ; oxidoreductase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A xylose reductase gene was isolated from the xylose-fermenting yeast Pachysolen tannophilus as a cDNA clone by selecting clones that hybridized specifically to xylose-inducible messenger RNA. Use of the cDNA clone as a probe in Northern hybridizations identified a xylose-inducible mRNA species large enough to encode a 36kDa xylose reductase protein known to be produced by this yeast. A corresponding genomic clone was isolated as a 3kb EcoRI fragment that specifically hybridized to the cDNA clone. The sequence of the cDNA and the largest open reading frame of the genomic clone are identical. The predicted translation product exhibits: (1) significant sequence identity with a previously published N-terminal amino acid sequence from purified P. tannophilus xylose (aldose) reductase protein exhibiting NADH/NADPH-dependent activities (aldose reductase, EC 1.1.1.21); (2) identity with a protein composed of 317 amino acid residues with a calculated molecular mass of 36·2kDa, equivalent to that reported for purified P. tannophilus xylose reductase; and (3) considerable sequence similarity to, and features of, a superfamily of oxidoreductases. This sequence is deposited as GenBank Accession Number U40706.
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    DNA-binding properties of the yeast transcriptional activator, Gcr1p (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 307-317 
    Details
    ISSN: 0749-503X
    Keywords: bent DNA ; Rap1p ; glycolytic enzyme gene-regulator ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In Saccharomyces cerevisiae the GCR1 gene product is required for high-level expression of genes encoding glycolytic enzymes. In this communication, we extend our analysis of the DNA binding properties of Gcr1p. The DNA-binding domain of Gcr1p binds DNA with high affinity. The apparent dissociation constant of the Gcr1p DNA-binding domain for one of its specific binding sites (TTTCAGCTTCCTCTAT) is 2·9×10-10 M. However, competition experiments showed that Gcr1p binds this site in vitro with a low degree of specificity. We measured a 33-fold difference between the ability of specific competitor and DNA of random sequence to inhibit the formation of nucleoprotein complexes between Gcr1p and a radiolabeled DNA probe containing its binding site. DNA band-shift experiments, utilizing probes of constant length in which the positions of Gcr1p-binding sites are varied relative to the ends, indicated that Gcr1p-DNA nucleoprotein complexes contain bent DNA. The implications of these findings in terms of the combinatorial interactions that occur at the upstream activating sequence elements of genes encoding glycolytic enzymes are discussed.
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  • 148
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    Sequence analysis of the 43 kb CRM1-YLM9-PET54-DIE2-SMI1-PHO81-YHB4-PFK1 region from the right arm of Saccharomyces cerevisiae chromosome VII (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 385-390 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VII ; sequence ; snRNa ; SNR7 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a 43 118 bp fragment from chromosome VII of Saccharomyces cerevisiae has been determined and analysed. The fragment originates from the right arm of chromosome VII. It starts approximately 11 kb centromere-proximal to the pet54 marker and ends in the middle of the PFK1 gene. The sequence contains a small nuclear RNA gene (SNR7) and 29 open reading frames (ORFs) larger than 100 amino acids. Six of these were completely internal to or partially overlapped other ORFs. Six previously described genes, YLM9/MRPL9, CRM1, DIE2, SMI1, PHO81 and YHB4, were mapped to this region in addition to pet54 and PFK1. Of the remaining 17 ORFs, four showed homology with other S. cerevisiae genes and four, including one of the partially overlapping ORFs, with genes from other organisms. Eight ORFs had no homology with any sequence in the databases. The actual sequences have been deposited in the EMBL database under Accession Number X87941.
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    Identification of a class of Saccharomyces cerevisiae mutants defective in fatty acid repression of gene transcription and analysis of the frm2 Gene (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 319-331 
    Details
    ISSN: 0749-503X
    Keywords: OLE1 ; lipids ; oleic acid ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Exogenous fatty acids transcriptionally control the expression of a wide variety of eukaryotic genes, many of which encode proteins involved in lipid metabolism. To identify gene products involved in the lipid signalling pathway, a reporter plasmid containing the 5′-upstream region of a gene demonstrated to be repressed by unsaturated fatty acids (OLE1) was fused in frame to the Escherichia coli gene lacZ encoding β-galactosidase. Saccharomyces cerevisiae mutants defective in transcriptional control by lipids were identified and this class of mutants has been named frm (fatty acid repression mutant). The mutants were organized into six complementation groups designated frm1-6. Mutants from two of the complementation groups, frm1 and frm3, were also defective in their ability to activate a reporter construct containing the 5′-upstream region of POX1. POX1 has been shown to be transcriptionally activated in the presence of unsaturated fatty acids. frm2 was rescued by a region of DNA localized to chromosome III. This region contained an open reading frame of 579 nucleotides predicted to encode a Mr 21 116 polypeptide. The upstream region of FRM2 contained a number of potential response elements which have previously been identified as important in regulating gene expression in response to glucose and certain fatty acids. Consistent with this observation, lacZ activity driven by FRM2 or frm2 promoters was induced two- to three-fold dependent upon the carbon and fatty acid source utilized. The properties of FRM2 suggest that it functions in the fatty acid signalling pathway and that it is itself regulated by fatty acids.
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    Candida maltosa NADPH-cytochrome P450 reductase: Cloning of a full-length cDNA, Heterologous expression in Saccharomyces cerevisiae and function of the N-terminal region for membrane anchoring and proliferation of the endoplasmic reticulum (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 333-348 
    Details
    ISSN: 0749-503X
    Keywords: protein transport ; high-level expression ; immunoelectron microscopy ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A full-length cDNA for NADPH-cytochrome P450 reductase from Candida maltosa was cloned and sequenced. The derived amino acid sequence showed a high similarity to the reductases from other eukaryotes.Expression in Saccharomyces cerevisiae under control of the GAL10 promoter resulted in an approximately 70-fold increase in NADPH-cytochrome c reductase activity in the microsomal fraction.The functional integrity of the heterologously expressed reductase as an electron transfer component for alkane hydroxylating cytochrome P450 from C. maltosa was shown in a reconstituted system containing both enzymes in a highly purified state. The signal-anchor sequence of the reductase was identified within the N-terminal region of the protein by means of constructing and expressing fusion proteins with the cytosolic form of yeast invertase. The first 33 amino acids turned out to be sufficient for stable membrane insertion, wild-type membrane orientation and retention in the endoplasmic reticulum.As shown by immunoelectron microscopy, the heterologously expressed reductase was integrated into the endoplasmic reticulum of the host organism. It triggered a strong proliferation of the membrane system. This membrane-inducing property of the reductase was transferable to the cytosolic reporter protein with the same N-terminal sequences that confer membrane insertion.The nucleotide sequence of the cDNA of NADPH-cytochrome P450 reductase from C. maltosa is available from the EMBL data library under Accession Number X76226.
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    Candida albicans homologue of GGP1/GAS1 gene is functional in Saccharomyces cerevisiae and contains the determinants for glycosylphosphatidylinositol attachment (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 361-368 
    Details
    ISSN: 0749-503X
    Keywords: Candida albicans ; Saccharomyces cerevisiae ; GPI anchor ; glycoproteins ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The GGP1/GAS1/CWH52 gene of Saccharomyces cerevisiae encodes a major exocellular 115 kDa glycoprotein (gp115) anchored to the plasma membrane through a glycosylphosphatidylinositol (GPI). The function of gp115 is still unknown but the analysis of null mutants suggests a possible role in the control of morphogenesis. PHR1 gene isolated from Candida alibicans is homologous to the GGP1 gene. In this report we have analysed the ability of PHR1 to complement a ggp1Δ mutation in S. cerevisiae. The expression of PHR1 controlled by its natural promoter or by the GGP1 promoter has been studied. In both cases we have observed a complete complementation of the mutant phenotype. Moreover, immunological analysis has revealed that PHR1 in budding yeast gives rise to a 75-80 kDa protein anchored to the membrane through a GPI, indicating that the signal for GPI attachment present in the C. albicans gene product is functional in S. cerevisiae.
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    Yeast protein translocation complex: Isolation of two genes SEB1 and SEB2 encoding proteins homologous to the Sec61β subunit (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 425-438 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; endoplasmic reticulum ; Sec61β ; translocation complex ; genetic interaction ; Seb1p/Sbh1p ; Seb2p ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A yeast gene (cDNA clone) was isolated in a screen for suppressors of secretion-defective sec15-1 mutation. This gene encodes a protein homologous to the β subunit of the mammalian Sec61 protein complex functioning in protein translocation into the endoplasmic reticulum (ER). The predicted protein, Seb1p, consists of 82 amino acids and contains one potential membrane-spanning region at the C-terminus but no N-terminal signal sequence. Seb1p shows 30% identity to the mammalian Sec61β subunit and 34% identity to the Arabidopsis thaliana Sec61β subunit. Overexpression of SEB1 from a multicopy plasmid suppressed the temperature sensitivity of sec61-2 and sec61-3 mutants. Immunofluorescence and immunoelectron microscopy indicated that Seb1p resides in the ER membranes with the hydrophilic N-terminus exposed to the cytoplasm. The in vitro translated Seb1p was post-translationally inserted into microsomal membranes. As the chromosomal disruption of the SEB1 gene was not lethal, potential homologous genes were screened by heterologous hybridization. The SEB1 homologue thus isolated, SEB2, encodes a protein 53% identical to Seb1p. Disruption of the chromosomal SEB2 was not lethal whereas the double disruption of SEB1 and SEB2 resulted in a temperature-sensitive phenotype. This study further emphasizes the evolutionary conservation of the ER protein translocation apparatus and provides novel genetic tools for its functional analysis. The sequences of SEB1 and SEB2 have been deposited in the EMBL database under Accession Numbers Z47789 and Z50012, respectively. The sequence of the Seb1 protein is identical to that of Sbh1p independently identified by Panzner et al. (Cell 81, 561-570. 1995) using a biochemical approach.
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    High-affinity glucose uptake in Saccharomyces cerevisiae is not dependent on the presence of glucose-phosphorylating enzymes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 439-447 
    Details
    ISSN: 0749-503X
    Keywords: transport ; glucose uptake ; Saccharomyces cerevisiae ; yeast ; rapid kinetics ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Glucose uptake in Saccharomyces cerevisiae is believed to consist of two kinetically distinguishable components, the affinity of which is modulated during growth on glucose. It has been reported that triple hexose-kinase deletion mutants do not exhibit high-affinity glucose uptake. This raises the question of whether and how high-affinity glucose uptake is related to the presence of glucose-phosphorylating enzymes. In this study the kinetics of glucose uptake in both wild-type cells and cells of hexose-kinase deletion mutants, grown on either glycerol or galactose, were determined using a rapid-uptake method. In wild-type cells glucose uptake measured over either 5 s or 200 ms exhibited high affinity. In contrast, in cells of hexose-kinase deletion mutants the apparent affinity of glucose uptake was dependent on the time scale during which uptake was measured. Measurements on the 5-s scale showed apparent low-affinity uptake whereas measurements on the 200-ms scale showed high-affinity uptake. The affinity and maximal rate of the latter were comparable to those in wild-type cells.Using a simple model for a symmetrical facilitator, it was possible to simulate the experimentally determined relation between apparent affinity and the time scale used.The results suggest that high-affinity glucose transport is not necessarily dependent on the presence of glucose-phosphorylating enzymes. Apparent low-affinity uptake kinetics can arise as a consequence of an insufficient rate of removal of intracellular free glucose by phosphorylation.This study underlines the need to differentiate between influences of the translocator and of metabolism on the apparent kinetics of sugar uptake in yeast.
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    Candida albicans phosphatidylinositol synthase has common features with both Saccharomyces cerevisiae and mammalian phosphatidylinositol synthases (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 449-456 
    Details
    ISSN: 0749-503X
    Keywords: inositol ; phosphatidylinositol synthase ; Candida albicans ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Phosphatidylinositol (PI) synthase (cytidine 5′-diphospho (CDP)-1,2-diacyl-sn-glycerol:myo-inositol 3phosphatidyltransferase, EC 2.7.8.11) was isolated from the microsomal cell fraction of Candida albicans. The Triton X-100 extracted enzyme was enriched 140-fold by affinity chromatography on CDP-diacylglycerol-Sepharose. The enzyme had a pH optimum at 9·5 in glycine/NaOH buffer. It had an absolute requirement for Mg2+ or Mn2+ and was inhibited by Ca2+ and Zn2+. Maximal activity was at 0·2-0·6 mm-CDP-diacylglycerol, higher concentrations inhibited the enzyme. With 2′-deoxy-CDP-diacylglycerol as the lipid substrate, optimal activity was at 0·7 mm. The Km for myo-inositol was determined to be 0·55 mm. The optimal temperature for the PI synthase reaction was 55°C. The C. albicans PI synthase shows differences to the Saccharomyces cerevisiae enzyme, such as activation by bivalent cations, inhibition by nucleotides, temperature optimum and activation energy, but also to the human PI synthase in preference for the lipid substrates, inhibition by nucleoside monophosphates and stabilization by Mn2+ and phospholipids.
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    The sequence of a 17 933 bp segment of Saccharomyces cerevisiae chromosome XIV contains the RHO2, TOP2, MKT1 and END3 genes and five new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 485-491 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XIV ; RHO2 ; TOP2 ; MKT1 ; END3 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the DNA sequence of a 17 933 bp fragment from the left arm of chromosome XIV of Saccharomyces cerevisiae. Analysis of the sequence reveals the presence of ten open reading frames (ORFs) larger than 100 codons. Four of these were previously identified as genes RHO2, TOP2, MKT1 and END3. Additionally, the NH2 end coding region of PMS1 is found in the 3′ end of the sequence. No significant homology to any known protein has been found for the other five ORFs. The nucleotide sequence has been deposited at EMBL, with Accession Number X89016.
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    The sequence of 12·8 kb from the left arm of chromosome XIV reveals a sigma element, a pro-tRNA and six complete open reading frames, one of which encodes a protein similar to the human leukotriene A4 hydrolase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 493-499 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; genome sequencing ; chromosome XIV ; leukotriene A4 hydrolase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the nucleotide sequence of a 12·8 kb fragment from the left arm of chromosome XIV carried by the cosmid 14-16d. An analysis of the sequence reveals the presence of a sigma element, a pro-tRNA gene and eight open reading frames, six of which are complete. All of the eight open reading frames correspond to new genes. Of the eight new genes, two show strong similarities to a pair of new genes from chromosome IX, suggesting an ancestral duplication, and one gene encodes a protein similar to mammalian leukotriene A4 hydrolase. The sequence has been deposited in the EMBL data bank under the Accession Number X94547.
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    The hsp150Δ-carrier confers secretion competence to the rat nerve growth factor receptor ectodomain in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 457-466 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; NGFR ; secretion ; protein production ; hsp150 protein ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: When the extracellular domain of rat low-affinity nerve growth factor receptor (NGFRe) was synthesized in Saccharomyces cerevisiae with the signal peptide of invertase, NGFRe was translocated to the endoplasmic reticulum (ER) and retained there. However, when NGFRe was fused to the C-terminus of the hsp150Δ-carrier, the hsp150Δ-NGFRe fusion protein was efficiently secreted to the growth medium with no apparent retention in the ER. The NGFRe portion was disulphide-bonded and its single N-glycosylation site was occupied. The hsp150Δ-carrier is an N-terminal signal peptide-containing fragment of a yeast secretory glycoprotein. Hsp150Δ-NGFRe, harvested from the culture medium, inhibited the cross-linking of [125I]NGF to authentic NGFR on the surface of human melanoma cells. Moreover, [125I]NGF could be chemically cross-linked to secretory hsp150Δ-NGFRe, suggesting that the NGFRe portion had adopted a ligand-binding conformation. However, inhibition of the cross-linking by unlabelled NGF was less effective than in the case of the authentic receptor. The hsp150Δ-carrier may have potential in the production of mammalian proteins, which require elaborate folding and disulphide formation in the ER.
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    Molecular cloning of a third chitinase gene (CHT1) from Candida albicans (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 501-504 
    Details
    ISSN: 0749-503X
    Keywords: Candida albicans ; chitinase ; yeast ; nucleotide sequencing ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Here we report the complete nucleotide sequence of a third chitinase gene (CHT1) from the dimorphic human pathogen Candida albicans. The deduced amino acid (aa) sequence of Cht1 consists of 416 aa and displays 36% protein sequence similarity to chitinases Cht2 and Cht3, from C. albicans. Interestingly the domain structure of Cht1 is truncated when compared to the other chitinases of C. albicans and lacks a Ser/Thr-rich region. The sequence data will appear in the GenBank Nucleotide Sequence Data Library under the accession number U36490.
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    Masthead (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320120501
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    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 515-522 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320120502
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    Factors affecting the mitotic stability of high-copy-number integration into the ribosomal DNA of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 467-477 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; ribosomal DNA ; plasmid ; mitotic stability ; heterologous protein ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Yeast vectors suitable for high-level expression of heterologous proteins should combine a high copy number with high mitotic stability. The pMIRY integrative vector system, based upon targeted integration into the yeast rDNA locus, developed in our laboratory satisfies these criteria. However, insertion of a (foreign) gene drastically reduced its mitotic stability of the resulting vector in comparison to its parent. In this paper we have investigated a number of possible reasons for this reduction in stability. The results demonstrate that plasmid size is an important, but not the only, determinant of mitotic stability. Stable maintenance is only observed when the complete plasmid has a size no larger than that of the rDNA unit (9·1 kb). In addition stability depends upon the nature of the rDNA fragment present in the plasmid, required for targeting its integration. On the other hand, it turned out to be irrelevant for mitotic stability whether the heterologous gene was expressed or not. These findings will be important in the design of a pMIRY vector suitable for industrial production of heterologous proteins.
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    The sequence of a 24 152 bp segment from the left arm of chromosome XIV from Saccharomyces cerevisiae between the BNI1 and the POL2 genes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 505-514 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XIV ; ORF analysis ; amino acid permeases ; polymerase II ; phosphatidylinositol-4-kinase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the framework of the European Union programme for sequencing the genome of Saccharomyces cerevisiae we have determined the nucleotide sequence of a region of 24 152 bp located on the left arm of chromosome XIV between the BNI1 and the POL2 genes. The sequence was obtained by directed sequence analysis using a mixture of ExoIII and primer walking strategies. Subsequent analysis revealed 13 open reading frames (ORFs) including four small ORFs completely internal to, or partly overlapping with, other ORFs. Five of these ORFs have been described previously (BNI1, APL1, LYP1, PIK1, POL2) and thus 74·8% of the 24 152 bp were already present in the databases prior to this sequencing effort. Interestingly, all 13 identified ORFs are characterized by a low codon adaptation index (0·04-0·22). In addition, this region of chromosome XIV shows an unusually high gene density with about 88% of coding DNA. This amounts to one gene per 2177 bp, which is significantly above the average gene length (about 1500 bp). For eight ORFs considerable homologies to ‘Expressed Sequence Tags’ derived from human cDNAs located in the XREF database could be identified. The complete nucleotide sequence of the 24 152 bp segment has been deposited in the EMBL data library under the Accession Number X92494.
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  • 163
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    Nucleotide sequences of alcohol acetyltransferase genes from lager brewing yeast, Saccharomyces carlsbergensis (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 593-598 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces carlsbergensis ; acetate ester ; alcohol ; acetyltransferase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequences of alcohol acetyltransferase genes isolated from lager brewing yeast, Saccharomyces carlsbergensis have been determined. S. carlsbergensis has one ATF1 gene and another homologous gene, the Lg-ATF1 gene. There was a high degree of homology between the amino acid sequences deduced for the ATF1 protein and the Lg-ATF1 protein (75·7%), but the N-terminal region has a relatively low degree of homology.Southern analysis and contour-clamped homogeneous electric field analysis of Saccharomyces strains suggest that the ATF1 gene is located on chromosome XV in S. cerevisiae and that the Lg-ATF1 gene might originate from the ‘non-S. cerevisiae’ genome of S. carlsbergensis, which is similar to that of S. bayanus and S. pastorianus. The nucleotide sequence data reported in this paper will appear in the DDBJ, EMBL and GenBank data banks with the Accession Numbers D63449 (ATF1) and D63450 (Lg-ATF1).
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  • 164
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    FEN2: A gene implicated in the catabolite repression-mediated regulation of ergosterol biosynthesis in yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 531-539 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; sterols ; fenpropimorph ; carbon catabolite repression ; nitrogen catabolite repression ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated and characterized a pleiotropic recessive mutation, fen2-1, that causes resistance to fenpropimorph and a low level of ergosterol in Saccharomyces cerevisiae. Ergosterol synthesis in the mutant strain was 5·5-fold slower than in the wild type; however, in vitro assays of the enzymes involved in ergosterol biosynthesis could not account for this low rate in the mutant. The mutant phenotype was expressed only in media exerting both carbon and nitrogen catabolite repression. To our knowledge, this is the first locus in yeast that reveals a concerted regulation between different pathways (carbon and nitrogen catabolite repression and/or general control of amino acid biosynthesis and ergosterol biosynthesis). The yeast gene FEN2 has been isolated and contains an open reading frame (ORF) of 512 codons. This ORF was found to be identical to YCR28C of chromosome III. A possible function of the FEN2 gene product in yeast is discussed.
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    The KNH1 gene of Saccharomyces cerevisiae is a functional homolog of KRE9 (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 683-692 
    Details
    ISSN: 0749-503X
    Keywords: S. cerevisiae ; β1,6-glucan synthesis ; duplicated enzymatic activity ; carbon-source-dependent transcriptional regulation ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The KNH1 gene from Saccharomyces cerevisiae was identified as an open reading frame on the right arm of chromosome IV. The product encoded by the KNH1 gene, Knh1p, shares 46% overall identity with Kre9p, a protein required for cell surface β1,6-glucan synthesis. While disruption of the KNH1 locus had no effect on cell growth, killer toxin sensitivity or β1,6-glucan levels, overexpression of KNH1 was found to suppress the severe growth defect of a kre9Δ mutant and restored the level of alkali-insoluble β1,6-glucan to almost wild-type levels. Knh1p, like Kre9p, can be found in the extracellular culture medium as an O-glycoprotein, with a molecular mass of 45-61 kDa. Disruption of both KNH1 and KRE9 is lethal, and unlike single kre9Δ mutants, could not be rescued by overproducing SKN7, a putative transcription factor involved in the regulation of extracellular matrix assembly. Transcription of KNH1 was found to be carbon-source and kre9Δ dependent, but SKN7 independent, suggesting that KNH1 is subject to alternative transcriptional control. The KNH1 sequence has been deposited in GenBank under Accession Number U31538.
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  • 166
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    Analysis of the DNA sequence of a 15,500 bp fragment near the left telomere of chromosome XV from Saccharomyces cerevisiae reveals a putative sugar transporter, a carboxypeptidase homologue and two new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 709-714 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; sugar transporter ; carboxypeptidase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence of a 15·5 kb DNA segment located near the left telomere of chromosome XV of Saccharomyces cerevisiae. The sequence contains nine open reading frames (ORFs) longer than 300 bp. Three of them are internal to other ones. One corresponds to the gene LGT3 that encodes a putative sugar transporter. Three adjacent ORFs were separated by two stop codons in frame. These ORFs presented homology with the gene CPS1 that encodes carboxypeptidase S. The stop codons were not found in the same sequence derived from another yeast strain. Two other ORFs without significant homology in databases were also found. One of them, O0420, is very rich in serine and threonine and presents a series of repeated or similar amino acid stretches along the sequence. The nucleotide sequence has been deposited in the EMBL data library under Accession Number X89715.
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    Synchronization affector of autonomous short-period-sustained oscillation of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 673-682 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; metabolic oscillation ; Synchronization affector ; continuous culture ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: When the yeast Saccharomyces cerevisiae was grown under aerobic continuous culture, an autonomous shortperiod-sustained oscillation appeared. This oscillation was observed in concentrations of various extracellular and intracellular parameters, such as ethanol, acetate, glycogen, dissolved oxygen and intracellular pH. In this work the synchronization affecter of this oscillation was investigated. Ethanol was found not to be the synchronizer of the oscillation because a pulse of ethanol did not affect the phase or period of the oscillation. The oscillation was dependent on the aeration rate, i.e., the oscillation occurred only between 150 and 600 ml min-1. However, the oxygen concentration did not influence synchronization as an upward shift in the oxygen concentration of the gas flow did not affect the sustainability of the oscillation. On the other hand, synchronization was stopped by an enhanced gas flow rate, keeping dissolved oxygen tension at the oscillatory condition, suggesting that synchronization was caused by a volatile compound in the culture. A stepwise increase in carbon dioxide concentration of the gas flow rate ceased synchronization, yet the oscillation seem to continue in each individual cell. Oscillatory behaviour of intracellular pH and carbon dioxide evolution rate showed a phase difference of 90 degrees. Based on these facts it is postulated that carbon dioxide, through the influence of its dissociation on intracellular pH, could be the synchronization affector of the autonomous short-period-sustained oscillation of S. cerevisiae.
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  • 168
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    The Candida albicans PKC1 gene encodes a protein kinase C homolog necessary for cellular integrity but not dimorphism (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 741-756 
    Details
    ISSN: 0749-503X
    Keywords: Candida albicans ; PKC1 gene ; Pkc1p activity ; cellular integrity ; dimorphism ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using a DNA fragment derived from the Saccharomyces cerevisiae protein kinase C gene (PKC1) as a probe to screen an ordered array library of genomic DNA from the dimorphic pathogenic fungus Candida albicans, the C. albicans PKC1 gene (CaPKC1) was isolated. The CaPKC1 gene is predicted to encode a protein of 1079 amino acids with 51% sequence identity over the entire length with the S. cerevisiae Pkc1 protein and is capable of functionally complementing the growth defects of a S. cerevisiae pkc1Δ mutant strain on hypo-osmotic medium. Deletion of both endogenous copies of the CaPKC1 gene in diploid C. albicans cells resulted in an osmotically remedial cell lysis defect of both the budding and the hyphal growth form and morphologically aberrant cells of the budding form. Despite these abnormalities, the transition between the two growth forms of C. albicans occurred normally in pkc1/pkc1 double disruptants. Capkc1p was modified at its C-terminus with two repeats of the Staphylococcus aureus protein A IgG-binding fragment (ZZ-sequence tag) and partially purified by chromatography on DEAE-Sepharose and IgG-Sepharose. In vitro, Capkc1p preferably phosphorylated the S. cerevisiae Pkc1p pseudosubstrate peptide and myelin basic protein, but not histones, protamine or dephosphorylated casein, and failed to respond to cofactors known to activate several mammalian PKC isozymes.
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  • 169
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    Green fluorescent protein as a marker for gene expression and subcellular localization in budding yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 773-786 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; functional analysis ; gene deletion ; green fluorescent protein ; homologous integration ; fusion protein ; expression vectors ; FACS ; microscopy ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The green fluorescent protein (GFP) from the jellyfish Aequorea victoria has attracted much attention as a tool to study a number of biological processes. This study describes the use of GFP as a vital reporter molecule for localization and expression studies in Saccharomyces cerevisiae. Construction of GFP expression vectors which allow N- or C-terminal fusion of the gfp gene to a gene of interest allowed the generation of fusion proteins whose subcellular localization was followed by fluorescence microscopy in living yeast cells. Analysis of three unknown open reading frames obtained from the budding yeast chromosome XIV resulted in distinct staining patterns, allowing prediction of the cellular localization of these unknown proteins. Furthermore, GFP was used to construct a gene replacement cassette which, after homologous integration into the genomic locus, placed the gfp gene behind a promoter of interest. The amount of GFP produced from this promoter was then quantified in living yeast cells by flow cytometry. With this novel replacement cassette a gene of interest can be deleted and at the same time its expression level studied under various growth conditions. The experiments presented here suggest that GFP represents a convenient fluorescent marker for localization studies as well as gene expression studies in budding yeast. Systematic studies of a large number of genes should benefit from such assays.
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  • 170
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    The isolation of a dol-P-man synthase from Ustilago maydis that functions in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 765-771 
    Details
    ISSN: 0749-503X
    Keywords: glycosylation ; dolichyl phosphoryl mannose ; dolichol ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genomic DNAs from several fungi were screened for a homologous sequence to Saccharomyces cerevisiae DPM1, an essential gene which encodes dolichyl phosphoryl mannose synthase. The fungi examined included Aspergillus nidulans, Neurospora crassa, Schizophyllum commune and Ustilago maydis. Only U. maydis gave a significant signal after Southern hybridization using DPM1 as a probe. The Ustilago homolog was subsequently cloned and sequenced. The predicted protein of 294 amino acids has 60% identity to the S. cerevisiae protein, but lacks the putative ‘dolichol recognition sequence’. RNA of ca. 900 bp is transcribed in both yeast and filamentous cells of Ustilago. In Escherichia coli, the U. maydis sequence expressed a 35 kDa protein exhibiting dolichyl phosphoryl mannose synthase activity. The sequence was also shown to complement a haploid strain of S. cerevisiae containing a deletion of the DPM1 gene. The U. maydis sequence therefore, encodes a dolichyl phosphoryl mannose synthase that can support normal vegetative growth in S. cerevisiae. The GenBank accession number is U54797.
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  • 171
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    Invertase secretion in Hansenula polymorpha under the AOX1 promoter from Pichia pastoris (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 815-822 
    Details
    ISSN: 0749-503X
    Keywords: HARS, Hansenula autonomously replicating sequence ; SUC2, sucrose invertase ; AOX1, alcohol oxidase ; Pollk, polymerase I, fragment klenow ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A DNA fragment containing a transcription regulating region of the alcohol oxidase (AOX1) gene from the methylotrophic yeast Pichia pastoris was used in the construction of a vector for the expression of heterologous proteins in the methylotrophic yeast Hansenula polymorpha. We used this vector to clone the SUC2 gene from Saccharomyces cerevisiae into H. polymorpha yeast.The culture conditions for invertase production using a fed-batch culture were studied. More than 1·5×103 U/ml of biologically active invertase (1 g/l) were secreted to the cellular periplasmic space. The fermentative process was scaled up to 50 l.Invertase produced from H. polymorpha was glycosylated, but it contained significantly less carbohydrate than protein produced by S. cerevisiae. Using the Western-blot technique, it was observed that invertase secreted from H. polymorpha and invertase secreted from S. cerevisiae showed common antigenic determinants.
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  • 172
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    Analysis of a 62 kb DNA sequence of chromosome X reveals 36 open reading frames and a gene cluster with a counterpart on chromosome XI (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 869-875 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome X ; open reading frames ; tRNA ; gene duplication ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a 61,989 bp stretch located between genes RAD7 and FIP1 of Saccharomyces cerevisiae chromosome X. This stretch contains 36 open reading frames (ORFs) of at least 100 codons. Fourteen of these correspond to sequences previously published as HIT1, CDC8, YAP17, CBF1, NAT1, RPA12, CCT5, TOR1, RFC2, PEM2, CDC11, MIR1, STE18 and GRR1. The proteins deduced from four ORFs (YJR059w, YJR065c, YJR075w, YJR078w) have significant similarity to proteins of known function from yeast or other organisms, including S. cerevisiae serine/threonine-specific protein kinase, Schizosaccharomyces pombe Act2 protein, S. cerevisiae mannosyltransferase OCH1 protein and mouse indoleamine 2,3-dioxygenase, respectively. Four of the remaining 18 ORFs have similarity to proteins with unknown function, six are weakly similar to other known sequences, while another eight exhibit no similarity to any known sequence. In addition, three tRNA genes have been recognized. Three genes clustered within 22 kb (YJR059w, YJR061w and TOR1) have counterparts arranged within 15 kb on the left arm of chromosome XI. The sequence has been deposited in the Genome Sequence Data Base under Accession Number L47993.
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    Sequence and analysis of a 33 kb fragment from the right arm of chromosome XV of the yeast Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 877-885 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome sequencing ; chromosome XV ; MGM1 ; STE4 ; CDC44 ; STE13 ; RPB8 ; MIP1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the nucleotide sequence of a cosmid (pEOA423) from chromosome XV of Saccharomyces cerevisiae. Analysis of the 33,173 bp sequence reveals the presence of 20 putative open reading frames (ORFs). Five of them correspond to previously known genes (MGM1, STE4, CDC44, STE13, RPB8). The previously published nucleotide sequences are in perfect agreement with our sequence except for STE4 and MGM1. In the latter case, 59 amino acids were truncated from the published protein at its N-terminal end due to a frameshift. The putative translation products of six other ORFs exhibit significant homology with protein sequences in public databases: O50 03 and O50 17 products are homologs of the ANC1 and MIP1 proteins of S. cerevisiae, respectively; O50 05 product is similar to that of a protein of unknown function from Myxococcus xanthus; O50 12 product is probably a new ATP/ADP carrier; O50 13 product shows homology with group II tRNA synthetases; and the O50 16 product exhibits strong similarity with the N-terminal domain of the NifU proteins from several prokaryotes. The remaining nine ORFs show no significant similarity. Among these, two contiguous ORFs (O50 19 and O50 20) are very similar to each other, suggesting an ancient tandem duplication. The 33,173 bp sequence has been submitted to EMBL (Accession Number: X92441).
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    Masthead (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320120901
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    Sticky-end polymerase chain reaction method for systematic gene disruption in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 859-868 
    Details
    ISSN: 0749-503X
    Keywords: gene disruption ; Saccharomyces cerevisiae ; yeast ; function analysis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe a new procedure for the generation of plasmids containing a large promoter and terminator region of a gene of interest, useful for gene disruption. In a two-step polymerase chain reaction (PCR), a fragment, corresponding to the terminator and promoter regions separated by a 16 bp sequence containing a rare restriction site (e.g. AscI), is synthesized (T-P fragment). This PCR fragment is cloned in vectors presenting a rare blunt-end cloning site and a yeast marker for selection in Saccharomyces cerevisiae (TRP1, HIS3 and KanMX). The final plasmids are used directly for gene disruption after linearization by the enzyme (e.g. AscI) specific for the rare restriction site. This approach was used to disrupt three open reading frames identified during the sequencing of COS14-1 from chromosome XIV of S. cerevisiae.
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    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 899-906 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320120902
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  • 177
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    A set of genetic markers for the chromosomes of the imperfect yeast Arxula adeninivorans (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1209-1217 
    Details
    ISSN: 0749-503X
    Keywords: Arxula adeninivorans ; yeast ; genetic markers ; linkage analysis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nuclear genome of the anamorphic yeast Arxula adeninivorans was analysed by benomyl-induced haploidization of parasexual hybrids marked with 32 auxotrophic mutations and pulsed field gel electrophoresis followed by DNA hybridization. Twenty-seven genes have been arranged into four linkage groups by haploidization, 15 genes belong to group 1, six to group 2, and three each to groups 3 and 4. Five genes could be localized by DNA hybridization on three out of four separated chromosomes. The gene LYS2 of the largest linkage group 1 and the 25S rDNA were identified on the largest chromosome, the GAA and the TEF1 gene on chromosome 2, and the ILV1 gene of linkage group 4 on the smallest chromosome.
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    Sequencing analysis of a 40·2 kb fragment of yeast chromosome X reveals 19 open reading frames including URA2 (5′ end), TRK1, PBS2, SPT10, GCD14, RPE1, PHO86, NCA3, ASF1, CCT7, GZF3, two tRNA genes, three remnant delta elements and a Ty4 transposon (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1471-1474 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome X ; retrotransposon ; delta element ; tRNA ; Ty4 ; URA2 ; TRK1 ; PBS2 ; SPT10 ; GCD14 ; RPE1 ; PHO86 ; NCA3 ; ASF1 ; CCT7 ; GZF3 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete sequence of a 40 247 bp DNA segment located on the left arm of chromosome X of Saccharomyces cerevisiae has been determined and analysed. The sequence encodes the 5′ coding region of the URA2 gene and 18 open reading frames of at least 100 amino acids. Ten of these correspond to known genes, whereas eight correspond to new genes. In addition, the sequence contains a tRNA-Ala gene, a tRNA-Asp gene, a Ty4 transposable element and three delta elements. The sequence has been deposited in the EMBL databank under Accession Numbers: Z49405, Z49404, Z49403, Z49402, Z49401, Z49400, Z49399, Z49398, Z49397, Z49396, Z49394, Z49392, Z49391, Z49390, Z49389, Z49387, Z49386, Z49385.
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    Nucleotide sequence analysis of a 40 kb segment on the right arm of yeast chromosome XV reveals 18 open reading frames including a new pyruvate kinase and three homologues to chromosome I genes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1475-1481 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; TEA1 ; RPA43 ; RPA190 ; SGC1 ; TYE7 ; REV1 ; PUT4 ; CIN1 ; MNE ; MRE4 ; tRNAPro ; tRNAMet ; YAL034 ; YAL037 ; Ty2 ; pyruvate kinase ; ubiquitin-conjugating enzyme ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the nucleotide sequence of a 40 kb fragment from the right arm of chromosome XV of Saccharomyces cerevisiae. Subsequent analysis revealed 18 non-overlapping open reading frames (ORFs) numbered from 06257 to 06357, an ARS, two tRNA genes and a Ty2 with its flanking elements. Ten ORFs have been sequenced previously: TEA1, RPA43, RPA190, SGC1 (also called TYE7) REV1, PUT4, CIN1, MNE and MRE4 (also called MEK1). Among the others, two seem to code for a new pyruvate kinase and for a new ubiquitin-conjugating enzyme; three have interesting homology with genes located on the left arm of chromosome I. This similarity with chromosome I extends to the left of the sequence presented here (Parle et al., submitted to Yeast). The homologous genes on the two chromosomes are placed in the same relative order. The complete nucleotide sequence of 40 295 bp and the deduced ORFs were submitted to the EMBL database (Accession Number X95720).
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    The SKS1 protein kinase is a multicopy suppressor of the snf3 mutation of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1407-1419 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; SKS1 ; protein kinase ; glucose transport ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Saccharomyces cerevisiae strains carrying snf3 are defective in high affinity glucose transport, and thus are unable to grow fermentatively on media with low concentrations of glucose. A multicopy suppressor of the snf3 growth defect, SKS1 (suppressor kinase of snf3), was found to encode a putative ser/thr protein kinase homologous to Ran1p, a kinase that regulates the switch between meiosis and vegetative growth in Schizosaccharomyces pombe. Overexpression of the SKS1 open reading frame is sufficient for suppression of the growth defects of snf3 mutants. Disruption of the open reading frame eliminates this suppression; as does the mutation of the consensus ATP binding site of Sks1p. A DDSE (DNA dependent snf3 suppressor element) was found to be present in the SKS1 promoter region. The suppression by this DDSE occurs in the absence of SKS1 coding region, that is, the DDSE can suppress a snf3 sks1 double null mutant which fails to grow fermentatively on low glucose as a snf3 mutant does. Both SKS1 and its DDSE can additionally suppress the growth defects of grr1 mutants, which are also impaired in high affinity glucose transport. The snf3 genomic suppressors, rgt1, RGT2 and ssn6, are also capable of suppressing snf3 associated growth defects in a strain lacking sks1.
    Additional Material: 5 Ill.
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  • 181
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    New vectors for combinatorial deletions in yeast chromosomes and for gap-repair cloning using ‘split-marker’ recombination (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1439-1457 
    Details
    ISSN: 0749-503X
    Keywords: yeast functional analysis ; gene disruption ; gap-repair cloning ; selection gene ‘pop-out’ ; I-SceI ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: New tools are needed for speedy and systematic study of the numerous genes revealed by the sequence of the yeast genome. We have developed a novel transformation strategy, based on ‘split-marker’ recombination, which allows generation of chromosomal deletions and direct gene cloning. For this purpose, pairs of yeast vectors have been constructed which offer a number of advantages for large-scale applications such as one-step cloning of target sequence homologs and combinatorial use. Gene deletions or gap-repair clonings are obtained by cotransformation of yeast by a pair of recombinant plasmids. Gap-repair vectors are based on the URA3 marker. Deletion vectors include the URA3, LYS2 and kanMX selection markers flanked by I-SceI sites, which allow their subsequent elimination from the transformant without the need for counter-selection. The application of the ‘split-marker’ vectors to the analysis of a few open reading frames of chromosome XI is described.
    Additional Material: 7 Ill.
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  • 182
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    The sequence of 55 kb on the left arm of yeast chromosome XVI identifies a small nuclear RNA, a new putative protein kinase and two new putative regulators (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1483-1492 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XVI ; REV3 ; SVS1 ; BEM4 ; CDC60 ; KIP2 ; PEP4 ; SPK1 ; KES1 ; SNR17B ; OYE3 ; RPL37A ; PAL1 (PXA1) ; leucine zipper ; protein kinase ; WD-40 repeats ; PTPA ; PRL1 ; PAR-1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced and analysed a 55 786 bp fragment located on the left arm of chromosome XVI of Saccharomyces cerevisiae. The sequence contains 29 non-overlapping open reading frames (ORFs) longer than 300 bp, among which 12 genes have previously been sequenced: OYE3, REV3, SVS1, BEM4, CDC60, KIP2, PEP4, SPK1, PAL1, KES1, SNR17B and RPL37A. Three new ORFs, P2591, P2594 and P2597 are highly homologous to the human phosphotyrosyl phosphatase activator PTPA, to the pleiotropic regulator PRL1 of PP1 and PP2a protein phosphatases in plants and to the protein kinase PAR-1 in Caenorhabditis elegans, respectively. Three other ORFs, P2545, P2567 and P2578 have significant homology with ORFs of unknown function located on yeast chromosomes VIII, XVI and IV respectively. The sequences in nucleotides and in amino acids have been deposited in the EMBL data library under the Accession Number X96770.
    Additional Material: 2 Ill.
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  • 183
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    Cloning and identification of HEM14, the yeast gene for mitochondrial protoporphyrinogen oxidase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1421-1425 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; protoporphyrinogen oxidase ; chromosome V ; functional analysis ; HEM14 ; YER014 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A respiratory-defective mutant (C54) of Saccharomyces cerevisiae was found to have a phenotype consistent with a mutation in either mitochondrial protoporphyrinogen oxidase or ferrochelatase. The mutant is grossly deficient in hemes, accumulates protoporphyrin and is rescued by exogenous heme. The increased levels of protoporphyrin at the expense of heme is indicative of a block in one of the two last steps of the heme biosynthetic pathway. Complementation of C54 by a known ferrochelatase mutant suggested that the defect was most likely in HEM14 encoding protoporphyrinogen oxidase. A plasmid capable of complementing C54 was obtained by transformation with a yeast genomic plasmid library. A partial sequence of the insert identified the gene as reading frame YER014 of yeast chromosome V (GenBank Accession Number U18778). This reading frame codes for a protein homologous to human protoporphyrinogen oxidase. Disruption of this gene elicits a respiratory defect and accumulation of protoporphyrin. The phenotype of the null mutant together with the homology of YER014p to human protoporphyrinogen oxidase provide compelling evidence that YER014 is HEM14.
    Additional Material: 3 Ill.
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  • 184
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    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1493-1500 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320121402
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  • 185
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    The yeast Rad6 protein: A mediator of homologous recombination across the scaffold attached region at the replication origin ARS1 (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1427-1438 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; ARS1 ; RAD6 ; scaffolds/SARs ; replacement recombination ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Here we show that the ubiquitin-conjugating enzyme Rad6p plays a crucial role in locus-specific replacement recombination in the TRP1-ARS1 region. In rad6-1 strains, where this ubiquitination activity is modified, homologous recombination across a 150 bp continuous region is completely abolished. Our results unambiguously identified the ARS1 scaffold attached region (SAR) as being the region where this impediment for replacement recombination is located, since a merging of the location of the recombination impediment and binding properties in a scaffold exchange assay with deletion mutations was observed. Our observations strongly support the notion of torsionally separated chromosomal domains being organized by SARs and scaffold proteins, and being dynamically realigned as a consequence of ubiquitination and proteolysis.
    Additional Material: 5 Ill.
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  • 186
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    Functional analysis of YCL09C: Evidence for a role as the regulatory subunit of acetolactate synthase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1511-1518 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; acetolactate synthase ; ILV6 ; EC 4.1.3.18 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have analysed the function of the open reading frame (ORF) YCL09C. The deletion of this ORF from chromosome III does not affect the physiology of the corresponding yeast strain enough to give a distinct phenotype. Nevertheless a computational analysis reveals high homology between this ORF and the enterobacterial genes encoding the regulatory subunit of acetolactate synthase. We have therefore tested the possibility that ycl09cp is the regulatory subunit of yeast acetolactate synthase by in vitro enzymatic analysis. The acetolactate synthase was previously shown to be retroinhibited by its final product valine. In Escherichia coli this retro-control is assured by the regulatory subunit. Using a yeast strain carrying a complete deletion of YCL09C, we have observed the loss of such retro-inhibition. These results together with the computational predictions show that YCL09C encodes the regulatory subunit of yeast acetolactate synthase.
    Additional Material: 5 Ill.
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  • 187
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    Analysis of a 26 756 bp segment from the left arm of yeast chromosome IV (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1549-1554 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome IV ; BPL1 ; RPB1 ; ARF2 ; L35 ; PPH21 ; rho GDP dissociation factor ; YL41B ; RGT2 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a 26·7 kb DNA segment from the left arm of Saccharomyces cerevisiae chromosome IV is presented. An analysis of this segment revealed 11 open reading frames (ORFs) longer than 300 bp and one split gene. These ORFs include the genes encoding the large subunit of RNA polymerase II, the biotin apo-protein ligase, an ADP-ribosylation factor (ARF 2), the ‘L35’-ribosomal protein, a rho GDP dissociation factor, and the sequence encoding the protein phosphatase 2A. Further sequence analysis revealed a short ORF encoding the ribosomal protein YL41B, an intron in a 5′ untranslated region and an extended homology with another cosmid (X83276) located on the same chromosome. The potential biological relevance of these findings is discussed. The sequence was submitted to the EMBL database under Accession Number X96876.
    Additional Material: 2 Ill.
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  • 188
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    Removal of an intron with unique 3′ branch site creates an amino-terminal protein sequence directing the scERV1 gene product to mitochondria (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1501-1510 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; intron splicing ; TACTAAC box ; mitochondrial localization ; mammalian homologues ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The yeast scERV1 gene is of special interest because it has a dual function in mitochondrial biogenesis and in the regulation of the cell cycle. The recent discovery that the yeast scERV1 gene has a structural and functional human homologue initiated a detailed comparison of the genes and their structures. In addition the homologous ALR (augmenter of liver regeneration) genes from rat and mouse have just been identified and it has been found that the mammalian proteins have a specific function in liver regeneration and in spermatogenesis. It now turns out that the organization of the 5′ regions of these genes is much more complicated than expected. The latest research has discovered an additional intron in the 5′ region of the mouse gene and possible amino-terminal extensions of the reading frames. In this work, reinvestigation of the 5′ region of the yeast gene identifies a putative intron with an unusual 3′ branch site. It is shown that a small intron of 83 nucleotides is present in this genomic region. Analysis of cDNA clones demonstrates that the intron is correctly removed from the messenger RNA and that therefore the unusual 3′ branch site is probably functional. Furthermore, studies with antibodies directed against recombinant scERV1 protein demonstrate that the gene product is associated with mitochondria, in agreement with its involvement in mitochondrial biogenesis. Complementation experiments with mutants and different 5′ deletions of the gene identify the corresponding promotor for transcription and the start codon for translation.
    Additional Material: 4 Ill.
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  • 189
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    Performance of industrial scale bioreactors with modified RUSHTON turbine agitators (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 43-56 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The data presented here with respect to the behaviour of industrial scale stirred tank bioreactors equipped with modified RUSHTON turbine agitators in the biosynthesis processes of antibiotics are valid for that case that the power consumption is the same as it is in standard RUSHTON turbine agitators.Each modified RUSHTON turbine agitator was obtained through the variation of the blade surface by adding perforations so that the ratio between the perforation surface area and the full surface area (or the surface fraction of the perforations) is 0.36.In the fermentations of Streptomyces aureofaciens, Streptomyces rimosus and Penicillium chrysogenum producing tetracycline, oxytetracyline and penicillin, respectively, in bioreactors equipped with modified RUSHTON turbine agitators, the relative antibiotic production is increased by more than 30% compared to standard bioreactors.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160107
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  • 190
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    Abbau industrieller Schadstoffe. Köln: Verlag TÜV Rheinland GmbH (Loseblatt-Ausgabe im Ordner); DM 198,-. ISBN 3-8249-0124-2 (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 42-42 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160106
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  • 191
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    Enhancement of the biological degradation of soils contaminated with oil by the addition of compost (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 19-30 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fundamentals of the biological treatment of contaminated soils were investigated in bioreactors with the aim to optimize the processes of biological soil treatment in order to achieve the highest possible degree of degradation within the shortest period of time. Preinvestigations using test systems at different scales have provided information about the possibilities of enhancing the decomposition processes which are dependent on various factors, such as milieu conditions, additives, etc., that must be known before remedial actions are taken.The investigations made so far have shown that compost is a favourable additive when oil-contaminated soils are biologically treated. The degradation of contaminants can be enhanced by the addition of compost. This positive effect is attributed to various mechanisms. In this paper, results of a variety of test systems at different scales are presented.In test series, different amounts of biocompost were added to investigate the influence on the degradation of diesel fuel. It was demonstrated that by increasing the compost content - the cumulative O2 consumption caused by the degradation of the diesel fuel contaminants increased.It could be shown that the reduction of the diesel fuel contaminants in the soil was independent of the compost age and amounted to approximately 94% of the initial quantity.The addition of biocompost could also enhance the degradation of real contaminants. After a test period of 162 days in set-ups with compost addition, more than 75% of the lubricating oil contaminants disappeared, while less than 37% of the contaminants disappeared in set-ups without compost addition.Moreover, by the addition of compost, the formation of pellets during the dynamic treatment of soil materials could be reduced.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160104
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  • 192
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    Media for the detection of Bacillus larvae spores in honey (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 57-64 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bacillus larvae, the causative agent of American foulbrood in honey bees completes its life cycle of germination, outgrowth and sporulation in young honey bee larvae by killing them and often bringing about the destruction of the entire hive. While B. larvae germinates and outgrows on complex organic media in vitro, the literature suggests, for reasons that are not at all clear, that a relatively large number of spores of B. larvae are required to yield each visible colony (colony forming units, CFU) on media. Various researchers have reported that from 16 to 3,000 or more spores of B. larvae are required to yield a single colony on an agar plate. HANSEN in Denmark designed a useful method of spreading approximately 80 mg of honey directly on the surface of a PETRI plate containing “J” agar medium to determine if B. larvae spores are present in the honey. In the present study, selected media were tested for the ability to recover B. larvae spores in honeys in the form of visible colonies (CFU) using HANSEN's strek method. A modification of a medium (TMYGP) developed by DINGMAN and STAHLY, (T-HCL-YGP agar), recovered considerably more viable B. larvae spores in the form of visible colonies (CFU) than HANSEN's “J” medium. When “J” medium was fortified with 0.1% sodium pyruvate, it was comparable to modified T-HCL-YGP medium in its recovery of B. larvae spores. Brain heart infusion agar (BHIA) with the addition of thiamine recovered more spores in the form of viable colonies than did “J” medium but it was not as efficient as T-HCL-YGP medium. Serial dilution from 100 to 10,000 times of weighed samples of honey with deionized water led to higher spore counts (CFU per g honey) than that indicated by undiluted honeys plated at 80 mg levels directly onto the surface of media by the HANSEN procedure.
    Additional Material: 2 Tab.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160108
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  • 193
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    Genetik. Grundlagen und Konzepte. Berlin, Heidelberg, New York, London, Paris, Tokyo, Hong Kong: Springer-Verlag, 1995 776 pages, 455 figures, 63 tables; DM 78,-ISBN 3-540-56075-0 (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 64-64 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160109
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  • 194
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    Relationship between citric acid and extracellular acid phosphatase production by Aspergillus niger (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 207-213 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Optimum conditions for citric acid production by Aspergillus niger in submerged culture were established. Some correlation was found between the ability of the fungus to produce citric acid and its capability to accumulate extracellular acid phosphatase. Mutagenization and passage on sodium citrate (10-25%) medium gave rise to eleven A. niger strains able to grow when the concentration was 25%. Two mutants showed an increase in citric acid production (8.8-25.7%), accompanied by the highest activity of acid phosphatase (43.1-46.6%).
    Additional Material: 3 Tab.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160221
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  • 195
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    Wir möchten zum. 9. Heidelberger Zytometrie Symposium einladen (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 80-80 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160114
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  • 196
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    Masthead (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996) 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160401
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  • 197
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    Recovery of bioproducts. EFB study report of the working party on down-stream processing. London: Society of Chemical Industry, 130 pages £ 25.00 (softcover). ISBN 0-901001-79-1 (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 236-236 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160403
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  • 198
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    Qualitätsmanagement im Gesundheitswesen. Köln: Verlag TÜV Rheinland, 1996 Ringhefter mit 1. Ergänzungslieferung; DM 148,00. ISBN 3-8249-0253-2 (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 245-246 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160405
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  • 199
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    PCR. Grundlagen und Anwendungen der Polymerase-Kettenreaktion. Stuttgart, Jena, New York: Gustav Fischer Verlag 123 pages, 27 figures, 5 tables; DM 44,-. ISBN 3-437-20509-9 (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 256-256 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160407
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  • 200
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    Substrate specificity of laccase from Lentinus edodes (1996)
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 257-270 
    Details
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: In previous studies, the white-rot basidiomycete Lentinus edodes, strain SC-495, was proved to be a “selective” lignin degrader and its extracellular crude preparations arising from solid-state cultures were successfully employed in biopulping experiments on annual plants. This fungus produced extracellular laccase as the predominant phenoloxidase when growing in solid-state fermentation on corn stalks. Laccase from this strain was purified and partially characterized, as an initial approach towards the study of its ligninolytic complex. Laccase was purified 69.6-fold by anion-exchange chromatography and two affinity-chromatography steps with an overall yield of 7.45%. The native enzyme exhibited a molecular mass of 74 kDa, an isoelectric point of 3.42 and a carbohydrate content of 7.5%. The absorption spectrum of laccase showed a maximum at 605 nm, typical of blue-copper oxidases. The optimum pH and temperature for the activity of laccase were 4.0-4.2 and 50°C, respectively. Kinetic experiments, performed with a wide range of phenolic compounds, showed that the reaction rate and the substrate affinity greatly varied depending on the nature of substituents and their reciprocal positions on the aromatic ring. In particular, the enzyme showed high affinity to phenolic compounds bearing methoxyl or methyl groups, but no affinity to those bearing the nitro group directly attached to the benzene ring, nor to non-phenolic lignin-related compounds, such as trans-cinnamic acid or 3,4-dimethoxycinnamic acid. The huge differences in terms of reactivity of the enzyme towards phenolic compounds suggests that a preliminary systematic screening should be advisable when using laccase in effluent treatment applications.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/abio.370160408
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