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  • 101
    Electronic Resource
    Electronic Resource
    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1385-1392 
    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.320121302
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  • 102
    Electronic Resource
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    Sequence of a 39 411 bp DNA fragment covering the left end of chromosome VII of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1555-1562 
    Details
    ISSN: 0749-503X
    Keywords: chromosome sequencing ; Saccharomyces cerevisiae ; ADH4 ; FZF1 ; HKB ; RTG2 ; HFM1 ; PDE1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a DNA fragment of 39 411 bp which includes part of the left telomere of chromosome VII of Saccharomyces cerevisiae. We have identified 19 open reading frames (ORFs); six correspond to known yeast genes (ADH4, FZF1, HKB, RTG2, HFM1 and PDE1), nine have similarity with other genes and four exhibit no significant similarity with any known gene. The average size of these ORFs seems to be related to their location, the eight ORFs nearest the telomere being shorter than the 11 others. These two groups of genes are separated by a region of 4·5 kb devoid of significant ORFs. One ORF, NRF120, is a new member of the seripauperine family, represented once in all sequenced yeast chromosomes, in a subtelomeric location. This sequence has been entered in the EMBL data library under accession number X94357.
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  • 103
    Electronic Resource
    Electronic Resource
    Analysis of a 22 956 bp region on the right arm of Saccharomyces cerevisiae chromosome XV (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1563-1573 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome XV ; ORFs ; predictable functions ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We present here the sequence analysis of a DNA fragment (cosmid pUOA1258) located on the right arm of chromosome XV. The 22 956 bp sequence reveals 14 open reading frames (ORFs) longer than 300 bp and the 201 bp RPS33 gene. Among the 14 large ORFs, two overlapping frames are likely to be non-expressed and one corresponds to the known GLN4 gene encoding glutaminyl-tRNA synthetase. Two ORFs, O3571 and O3620, encode putative transcriptional regulators with a Zn(2)-Cys(6) DNA binding domain characteristic of members of the GAL4 family. Among the nine remaining ORFs, five (O3568, O3575, O3590, O3615 and O3625) present significant similarity to proteins of unknown function and four (O3580, O3595, O3630 and O3635) lack homology to sequences present in the databases screened. This sequence has been deposited in the GenBank database under Accession Number U55021.
    Additional Material: 6 Ill.
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  • 104
    Electronic Resource
    Electronic Resource
    Analysis of a 23 kb region on the left arm of yeast chromosome IV (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1587-1592 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; chromosome IV ; Saccharomyces cerevisiae ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the complete nucleotide sequence of a 23 kb segment from the left arm of chromosome IV, which is carried by the cosmid 1L10. This sequence contains the 3′ coding region of the STE7 and RET1 (COP1) genes, and 13 complete open reading frames longer than 300 bp, of which ten correspond to putative new genes and three (CLB3, MSH5 and RPC53) have been sequenced previously. The sequence from cosmid 1L10 was obtained entirely by a combined subcloning and walking primer strategy.
    Additional Material: 4 Ill.
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  • 105
    Electronic Resource
    Electronic Resource
    Sequence and analysis of a 26·9 kb fragment from chromosome XV of the yeast Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1575-1586 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome sequencing ; chromosome XV ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the nucleotide sequence of a fragment of chromosome XV of Saccharomyces cerevisiae cloned into cosmid pEOA048. The analysis of the 26 857 bp sequence reveals the presence of 19 open reading frames (ORFs), and of one RNA-coding gene (SNR17A). Six ORFs correspond to previously known genes (MKK1/SSP32, YGE1/GRPE/MGE1, KIN4/KIN31/KIN3, RPL37B, DFR1 and HES1, respectively), all others were discovered in this work.Only five of the new ORFs have significant homologs in public databases, the remaining eight correspond to orphans (two of them are questionable). O5248 is a probable folylpolyglutamate synthetase, having two structural homologs already sequenced in the yeast genome. O5273 shows homology with a yeast protein required for vanadate resistance. O5268 shows homology with putative oxidoreductases of different organisms. O5257 shows homology with the SAS2 protein and another hypothetical protein from yeast. The last one, O5245, shows homology with a putative protein of Caenorhabditis elegans of unknown function. The present sequence corresponds to coordinates 772 331 to 799 187 of the entire chromosome XV sequence which can be retrieved by anonymous ftp (ftp. mips. embnet. org).
    Additional Material: 5 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.320121501
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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. 1593-1600 
    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.320121502
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  • 108
    Electronic Resource
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    Identification of proteins of the yeast protein map using genetically manipulated strains and peptide-mass fingerprinting (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1519-1533 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces ; yeast protein map ; protein identification ; mass spectrometry ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this study we used genetically manipulated strains in order to identify polypeptide spots of the protein map of Saccharomyces cerevisiae. Thirty-two novel polypeptide spots were identified using this strategy. They corresponded to the product of 23 different genes. We also explored the possibilities of using peptide-mass fingerprinting for the identification of proteins separated on our gels. According to this strategy, proteins contained in spots are digested with trypsin and the masses of generated peptides are determined by matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS). The peptide masses are then used to search a yeast protein database for proteins that match the experimental data. Application of this strategy to previously identified polypeptide spots gave evidence of the feasibility of this approach. We also report predictions on the identities of nine unknown spots using MALDI-MS.
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  • 109
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    A novel cell wall protein specific to the mycelial form of Yarrowia lipolytica (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1535-1548 
    Details
    ISSN: 0749-503X
    Keywords: Yarrowia lipolytica ; cell wall ; mycelium ; cDNA ; YWP1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A cDNA clone specifying a cell wall protein was isolated from a Yarrowia lipolytica cDNA library. The cDNA library was constructed in the expression vector λgt11, with the RNA isolated from actively growing mycelial cells. The deduced amino acid sequence shows that the encoded protein contains an N-terminal hydrophobic signal peptide. We have designated this protein YWP1 for Yarrowia lipolytica cell Wall Protein. Northern hybridization identified YWP1 transcript only when Y. lipolytica was growing in the mycelial form. The encoded protein seems to be covalently bound to the glucan cell wall since it is not released from the cell walls by sodium dodecyl sulphate extraction, but it is solubilized following partial degradation of β-glucan by Zymolyase digestion. The protein is localized in the outer surface on the tip of the growing mycelial cells and is found partially cryptic in sub-apical locations, suggesting that it participates directly in the mycelial wall architecture.
    Additional Material: 7 Ill.
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  • 110
    Electronic Resource
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    Publisher's note (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1601-1601 
    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.320121602
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  • 111
    Electronic Resource
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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.320121601
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  • 112
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    List of referees (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1602-1602 
    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.320121603
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  • 113
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    The DNA sequence of cosmid 14-5 from chromosome XIV reveals 21 open reading frames including a novel gene encoding a globin-like domain (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1071-1076 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; genome sequencing ; SLA2 ; ZWF1 ; BLH1 ; KEX2 ; SIN4 ; URE2 ; globin ; DnaJ/kw ; zinc-finger ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this paper is described the DNA sequence of cosmid 14-5 from chromosome XIV of Saccharomyces cerevisiae. The sequence is 38 855 bases long and contains 21 open reading frames (ORFs) plus three internal ORFs. Six ORFs correspond to known yeast genes (SLA2, ZWF1, BLH1, KEX2, SIN4 and URE2); two other ORFs had already been sequenced because they are adjacent to known genes; the remaining 12 ORFs are novel genes. Of these, one ORF (N1142) is particularly interesting since it shows a significant similarity to mammalian globin. Another ORF (N1254) displays two zinc finger motifs as well as a DNAJ motif. The cosmid sequence has been submitted to the EMBL data library under Accession Number Z69381.
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  • 114
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    The sequence of a 20·3 kb DNA fragment from the left arm of Saccharomyces cerevisiae chromosome IV contains the KIN28, MSS2, PHO2, POL3 and DUN1 genes, and six new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1077-1084 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; genome sequencing ; chromosome IV ; KIN28 ; MSS2 ; PHO2 ; POL3 ; DUN1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence of a 20 300 bp DNA fragment from the left arm of Saccharomyces cerevisiae chromosome IV. This segment contains 13 complete open reading frames (ORFs) and part of another ORF, altogether covering 84·2% of the entire sequence, five of which correspond to the previously characterized KIN28, MSS2, PHO2, POL3/CDC2 and DUN1 genes. One putative protein, D2358p, shares considerable homology with an O-sialoglycoprotein endopeptidase from Pasteurella haemolytica serotype A1. The putative product of D2325 contains the characteristic consensus motif of triacylglycerol lipases. D2320p and D2352p have a putative ‘leucine-zipper’ structure and a RNA-binding region Rnp-1 signature, respectively. The sequence data have been submitted to EMBL data library under Accession Number X95644.
    Additional Material: 3 Ill.
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  • 115
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    The sequence of a 30 kb fragment on the left arm of chromosome XV from Saccharomyces cerevisiae reveals 15 open reading frames, five of which correspond to previously identified genes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1091-1095 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; genome sequencing ; PHO80 ; TIR2 ; SLG1 ; ATP-dependent permease ; subtilisin-like protease III ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence of a 30 469 bp long DNA fragment on the left arm of chromosome XV of Saccharomyces cerevisiae. The fragment contains 15 open reading frames (ORFs) of at least 300 bp. Five previously sequenced yeast genes, PHO80, TIR2, SLG1, the gene encoding the subtilisin-like protease III precursor and the gene coding for ATP-dependent permease, are found among these ORFs. By DNA sequence comparison, two ORFs identified previously reported expressed sequence tags from yeast. Of the proteins encoded by the remaining eight ORFs, six show similarities to proteins from different organisms and two lack detectable similarity with any amino acid sequence described in public data banks. The DNA sequence has been deposited in GenBank under Accession Number U43491.
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  • 116
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    Sequencing and analysis of a 35·4 kb region on the left arm of chromosome IV from Saccharomyces cerevisiae reveal 23 open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1085-1090 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome IV ; SNQ2 ; SES1 ; GCV1 ; RPL2B ; HEX2/SRN1 ; RPS18A ; tRNA-Val12a ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete DNA sequence of cosmid clone 31A5 containing a 35 452 bp segment from the left arm of chromosome IV from Saccharomyces cerevisiae, was determined from an ordered set of subclones in combination with primer walking on the cosmid. The sequence contains 23 open reading frames (ORFs) of more than 100 amino acid residues and the tRNA-Val2a gene. Five ORFs corresponded to the known yeast genes SNQ2, SES1, GCV1, RPL2B and RPS18A. The DNA sequence for RPS18A is interrupted by an intron. One ORF corresponded to a part of the yeast gene HEX2 at the end of the cosmid insert. Four ORFs encoded putative proteins which showed strong homologies to other previously known proteins, three of yeast origin and one of non-yeast origin. Two ORFs were classified as having borderline homologies: one had similarity to two protein families and another to two protein products of unknown function from other species. The remaining 11 ORFs bore no significant similarity to any published protein. The complete DNA sequence has been submitted to the EMBL data library, Accession Number X95966.
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  • 117
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    Sequence of 29 kb around the PDR10 locus on the right arm of Saccharomyces cerevisiae chromosome XV: Similarity to part of chromosome I (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 999-1004 
    Details
    ISSN: 0749-503X
    Keywords: MYO2 ; SNC2 ; PDR10 ; SCD5 ; FTB1 ; MIP1 ; VMA4 ; MRS2 ; ALA1 ; KRE5 ; TEA1 ; YAL034c ; duplicate chromosomal region ; ABC transporter ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report a 29,445 bp sequence from the right arm of yeast chromosome XV. It contains the genes MYO2, SNC2, PDR10, SCD5 (also called FTB1), MIP1, VMA4, MRS2, ALA1, KRE5, TEA1, and a homologue of YAL034c. Several discrepancies with previously published sequences were found. PDR10 encodes a protein highly similar to the pleiotropic drug resistance protein Pdr5p. This sequence contig forms part of a region of extended similarity to part of the left arm of chromosome I, which is a relic of an ancient duplicated chromosomal region.
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  • 118
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    Sequencing analysis of a 4·1 kb subtelomeric region from yeast chromosome IV identifies HXT15, a new member of the hexose transporter familyM.B. and D.S. contributed equally to his paper. (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1005-1011 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; yeast ; chromosome IV ; HXT15 ; hexose transporter ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The DNA sequence of a 4·1 kb region of Saccharomyces cerevisiae chromosome IV was determined. This region contains a single open reading frame which codes for a member of the hexose transporter family. This new gene has been named HXT15 according to yeast gene data bases. The sequence has been entered in the EMBL data library under Accession Number X92891.
    Additional Material: 4 Ill.
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  • 119
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    Sequencing of a 35·71 kb DNA segment on the right arm of yeast chromosome XV reveals regions of similarity to chromosomes I and XIIIMS dedicates this paper to the memory of his friend and colleague Angelos Kalogeropoulos who died on April 19th 1996. (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1021-1031 
    Details
    ISSN: 0749-503X
    Keywords: SLY41 ; SPS4 ; COT1 ; FAA1 ; PMT3 ; PRO2 ; MYO2 ; EST sequence ; intron-containing ORFs ; transmembrane domains ; RNA-binding domains ; duplicated chromosomal region ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In a shotgun approach we sequenced the cosmid pEOA284 containing a fragment derived from the right arm of chromosome XV of Saccharomyces cerevisiae. An analysis of the sequence revealed that it contained open reading frames (ORFs) corresponding to the known genes SLY41, SPS4, COT1, FAA1, PMT3, PRO2 and MYO2. Of the 18 unknown ORFs, five are contained totally within, and two, O6105 and O6163, partially overlap other ORFs. ORF O6116 and O6139 have putative introns. Regions of similarity with chromosomes I and XIII have been uncovered. Interestingly, most of the paired ORFs encode proteins of the same gene family. The relatedness of these ORFs suggests gene duplication. The sequence has been entered into the public data libraries under Accession Number X90565.
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  • 120
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    Sequence analysis of a 13·4 kbp fragment from the left arm of chromosome XV reveals a malate dehydrogenase gene, a putative Ser/Thr protein kinase, the ribosomal L25 gene and four new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1013-1020 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; MDH2 gene ; Ser/Thr protein kinases ; ribosomal genes ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A 13 421 bp fragment located near the left telomere of chromosome XV (cosmid pEOA461) has been sequenced. Seven non-overlapping open reading frames (ORFs) encoding polypeptides longer than 100 residues have been found (AOB859, AOC184, AOE375, AOX142i, AOE423, AOA476 and AOE433). An additional ORF (AOE131) is found within AOA476. Three of them (AOC184, AOA476 and AOE433) show no remarkable identity with proteins deposited in the data banks. ORF AOB859 is quite similar to a hypothetical yeast protein of similar size located in chromosome VI, particularly within the C-terminal half. AOE375 encodes a new member of the glycogen synthase kinase-3 subfamily of Ser/Thr protein kinases. AOX142i is the gene encoding the previously described ribosomal protein L25. AOE423 codes for a protein virtually identical to the MDH2 malate dehydrogenase isozyme. However, our DNA sequence shows a single one-base insertion upstream of the reported initiating codon. This would produce a larger ORF by extending 46 residues the N-terminus of the protein. The existence of this insertion has been confirmed in three different yeast strains, including FY1679. The complete nucleotide sequence of the 13·4 kbp fragment has been deposited at the DNA databases (Accession Number U41293).
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  • 121
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    DNA sequence analysis of a 10 624 bp fragment of the left arm of chromosome XV from Saccharomyces cerevisiae reveals a RNA binding protein, a mitochondrial protein, two ribosomal proteins and two new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1041-1045 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; ribosomal proteins ; RNA binding protein ; mitochondrial protein ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the sequence of a 10 624 bp DNA segment located in the left arm of chromosome XV of Saccharomyces cerevisiae. The sequence contains eight open reading frames (ORFs) longer than 100 amino acids. Two of them do not present significant homology with sequences found in the databases. The product of ORF o0553 is identical to the protein encoded by the gene SMF1. Internal to it there is another ORF, o0555 that is apparently expressed. The proteins encoded by ORFs o0559 and o0565 are identical to ribosomal proteins S19.e and L18 respectively. ORF o0550 encodes a protein with an RNA binding signature including RNP motifs and stretches rich in asparagine, glutamine and arginine. The nucleotide sequence determined has been deposited in the EMBL data library under Accession Number X95258.
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  • 122
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    A putative helicase, the SUA5, PMR1, tRNALys1 genes and four open reading frames have been detected in the DNA sequence of an 8·8 kb fragment of the left arm of chromosome VII of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1033-1040 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome VII ; ROK1 ; PMR1 ; SUA5 ; tRNALys1 ; ATP-dependent RNA helicase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence of an 8·8 kb segment of DNA from the left arm of chromosome VII of Saccharomyces cerevisiae. The sequence reveals seven open reading frames (ORFs) G1651, G1654, G1660, G1663, G1666, G1667 and G1669 greater than 100 amino acids in length and the tRNALys1 gene. ORF G1651 shows 100% identity with the ROK1 protein which is a putative RNA helicase of the ‘DEAD box’ protein family. ORF G1654 exhibits a motif highly conserved in ATP/GTP binding proteins generally referred to as ‘P-loop’. From FastA analysis, G1660 and G1666 were found to be previously sequenced genes, respectively SUA5 and PMR1. The three other ORFs identified are partially (G1663) or completely (G1667 and G1669) overlapping with the PMR1 sequence on the complementary strand. This feature, together with their low codon adaptation indexes and the absence of significant homology with known proteins suggest that they do not correspond to real genes. The nucleotide sequence of the 8·8 kb fragment is available through the EMBL data library under the Accession Number X85757.
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  • 123
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    Identification of a putative methylenetetrahydrofolate reductase by sequence analysis of a 6·8 kb DNA fragment of yeast chromosome VII (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1047-1051 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome VII ; methylenetetrahydrofolate reductase ; SCS3 ; SUP44 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence analysis of a 6·8 kb DNA fragment from Saccharomyces cerevisiae chromosome VII. This sequence contains five open reading frames (ORFs) greater than 100 amino acids. There is also an incomplete ORF flanking one of the extremes, G2868, which is the 3′ end of the SCS3 gene (Hosaka et al., 1994). The translated sequence of ORF G2882 shows similarity to the human methylenetetrahydrofolate reductase (Goyette et al., 1994). ORF G2889 shows no significant homologies with the sequences compiled in databases. ORF G2893 corresponds to the gene SUP44, coding for the yeast ribosomal protein S4 (All-Robin et al., 1990). G2873 and G2896 are internal ORFs. The whole sequence of the fragment is available at the EMBL nucleotide sequence database, GenBank and Data Bank of Japan under the Accession Number X94106.
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  • 124
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    Sequence analysis of a 12 801 bp fragment of the left arm of yeast chromosome XV containing a putative 6-phosphofructo-2-kinase gene, a gene for a possible glycophospholipid-anchored surface protein and six other open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1053-1058 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; 6-phosphofructo-2-kinase ; glycophospholipid-anchored surface protein ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The DNA sequence of a 12,801 bp fragment located near the left telomere of chromosome XV has been determined. Sequence analysis reveals eight open reading frames (ORFs) encoding polypeptides larger than 100 residues. ORFs AOE129 and AOAA121 are in opposite strands and they overlap at their 3′ ends. AOE397 has similarity with phosphofructokinase genes from other organisms and may code for a second 6-phosphofructo-2-kinase of Saccharomyces cerevisiae. Sequence of AOA471 shows significant similarity with yeast genes coding for glycophospholipid-containing proteins. AOD1341 would code for a 1341 amino acids long protein with a predicted ATP/GTP-binding site and a transmembrane domain. The nucleotide sequence reported here has been submitted to the EMBL data library under Accession Number X95465.
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  • 125
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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.320121101
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  • 126
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    DNA sequence analysis of the VPH1-SNF2 region on chromosome XV of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1059-1064 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; Saccharomyces cerevisiae ; chromosone XV ; DNA ; VPH1 ; PAC1 ; MOD5 ; CAP20 ; ORF1 ; SNF2 ; DFR1 ; DHFR ; heat shock protein ; protein disulfite isomerase ; tRNA-ala ; sigma ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a 37 000 base pair region from the left arm of chromosome XV of Saccharomyces cerevisiae has been determined and analysed. This region contains 21 open reading frames (ORFs) coding for proteins of more than 100 amino acids. Six ORFs correspond to the genes PAC1, VPH1, MOD5, CAP20, ORF1 and SNF2 already described. Eight ORFs show some similarities to known genes from yeast and other organisms. They include genes coding for serine/threonine protein kinases, a multidrug resistance family homologue, a protein related to dihydrofolate reductase, a cluster of heat shock-like proteins and a gene coding for an enzyme related to protein disulfide isomerase. Finally seven ORFs do not show any similarities with a known gene. In addition we found a new ala-tRNA (UGC) gene located next to a sigma sequence. The sequence has been deposited in the EMBL databank under Accession Number X89633.
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    The sequence of 23 kb surrounding the SNF3 locus on the left arm of yeast chromosome IV reveals the location of five known genes and characterizes at least six new open reading frames including putative genes for ribosomal protein L35 and a sugar transport protein (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1065-1070 
    Details
    ISSN: 0749-503X
    Keywords: MGT1 ; SHM1 ; ASF2 ; WEB1 ; SNF3 ; ARF1 ; L35 ribosomal protein ; sugar transport protein ; Saccharomyces carlsbergensis, sake, diastaticus ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of 22,846 bp of the left arm of chromosome IV is described. Twelve open reading frames (ORFs) greater than 100 triplets were detected, one of which extends into an adjacent cosmid. Two of the ORFs may contain an intron. One of these is an L35 ribosomal protein gene. Five ORFs (D1204, D1214, D1219, D1234 and D1244) encode previously sequenced genes (MGT1, SHM1, ASF2, SNF3 and ARF2, respectively). The nucleotide sequence of a sixth ORF (D1229) is quite similar to the WEB1 gene, which appeared in the DNA databases shortly after finishing the sequence reported here. It is not clear whether or not WEB1 and D1229 represent one and the same gene. The co-linearity of the reported DNA sequences with the genome of strains from Saccharomyces cerevisiae subspecies carlsbergensis, sake and diastaticus was assessed by comparative PCR with overlapping primer sets. The nucleotide sequence data reported in this paper will appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under Accession Number X83276. © 1996 John Wiley & Sons, Ltd.
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  • 128
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    Sequence analysis of a 14·2 kb fragment of Saccharomyces cerevisiae chromosome XIV that includes the ypt53, tRNALeu and gsr m2 genes and four new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 599-608 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XIV ; ypt53 ; tRNALeu ; gsr m2 ; S7 RPR ; transmembrane protein ; norA ; glutamic acid rich protein ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: As part of the EU yeast genome program, a fragment of 14 262 bp from the left arm of Saccharomyces cerevisiae chromosome XIV has been sequenced. This fragment corresponds to cosmid 14-14b and is located roughly 130 kb from the centromere. It contains four new open reading frames which encode potential proteins of more than 99 amino acids, as well as the ypt53, tRNALeu and gsr m2 genes. The putative protein N2212 is similar to the ribosomal protein S7 from humans. N2215 contains several predicted transmembrane elements. N2231 contains regions which are rich in acidic, as well as basic, residues which could form α-helical structures. Similar regions are found in a variety of proteins including glutamic acid rich protein, trichohyalin, caldesmon, Tb-29 and several cytoskeleton-interacting proteins. The sequence has been entered in the EMBL data library under Accession Number X85811.
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  • 129
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    Isolation and characterization of Schizosaccharomyces pombe fragile mutants (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 555-564 
    Details
    ISSN: 0749-503X
    Keywords: Schizosaccharomyces pombe ; fragile mutants ; cell wall structure ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Three Schizosaccharomyces pombe fragile mutants requiring the presence of an osmotic stabilizer to grow, that lyse when transferred into hypotonic solutions and that secrete to the extracellular medium more protein than the parental strain were isolated. In the three mutants, the fragile phenotype segregated in a Mendelian fashion, indicating a single chromosomal gene mutation, and behaved as a recessive character. By complementation analysis, the three fragile mutants fell in a single complementation group, defining the same gene (SRB1). Mutations of this gene are responsible for alterations in the cells such as fragile character, increase in the cell wall porosity, changes in the cell morphology and floc-forming ability. The study of the three srb1 alleles indicated that the degree of these alterations is proportional to a significant decrease in the galactomannan fraction of the mutants cell wall. The data presented in this report suggest that the product of the SRB1 gene is critical for the maintenance of the integrity and structure of Sz. pombe cell wall.
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    ERG1, encoding squalene epoxidase, is located on the right arm of chromosome VII of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 609-613 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; ERG1 ; squalene epoxidase ; chromosome VII ; sterol biosynthesis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ERG1 gene of Saccharomyces cerevisiae encodes squalene epoxidase, a key enzyme in the ergosterol pathway. ERG1 is an essential gene. Disruption of the gene with URA3 results in a lethal phenotype when cells are grown under aerobic conditions, even in the presence of ergosterol. However, cells are viable in the presence of ergosterol under anaerobic growth conditions during which ergosterol is taken up by cells. Physical and genetic mapping data reveal that ERG1 is located on the right arm of chromosome VII proximal to QCR9 at a distance of 14·6 cM from ADE3.
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  • 131
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    PetCR46, a gene which is essential for respiration and integrity of the mitochondrial genome (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 577-582 
    Details
    ISSN: 0749-503X
    Keywords: Chromosome III ; YCR46C ; nuclear petite ; mitochondrial DNA ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the frame of the European Pilot Project for the functional analysis of newly discovered open reading frames (ORFs) from Saccharomyces cerevisiae chromosome III, we have deleted entirely the YCR46C ORF by a one-step polymerase chain reaction method and replaced it by the HIS3 marker in the strain W303. The deletion has been checked by meiotic segregation and Southern blot analyses. Characterization of the deleted strain indicates that YCR46C is essential for respiration and maintenance of the mitochondrial genome since its deletion leads to the appearance of 100% of cytoplasmic petites. Hybridization with molecular probes from mtDNA of individual clones of such petites showed that about 50% did hybridize (rho- clones) while others did not (possibly rho° clones). The wild-type gene has been cloned and shown to complement the deletion. The gene, which probably codes for a mitochondrial ribosomal protein, has been called petCR46.
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  • 132
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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.320120601
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  • 133
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    Review: The Cct eukaryotic chaperonin subunits of Saccharomyces cerevisiae and other yeasts (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 523-529 
    Details
    ISSN: 0749-503X
    Keywords: chaperonin ; Cct complex ; protein folding ; Candida albicans ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: All eight of the CCT1-CCT8 genes encoding the subunits of the Cct chaperonin complex in Saccharomyces cerevisiae have been identified, including three that were uncovered by the systematic sequencing of the yeast genome. Although most of the properties of the eukaryotic Cct chaperonin have been elucidated with mammalian systems in vitro, studies with S. cerevisiae conditional mutants revealed that Cct is required for assembly of microtubules and actin in vivo. Cct subunits from the other yeasts, Candida albicans and Schizosaccharomyces pombe, also have been identified from partial and complete DNA sequencing of genes. Cct8p from C. albicans, the only other completely sequenced Cct protein from a fungal species other than S. cerevisiae, is 72% and 61% similar to the S. cerevisiae and mouse Cct8 proteins, respectively. The C. albicans CCT8 sequence has been assigned the Accession Number U37371 in the GenBank/EMBL database.
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    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 615-622 
    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.320120602
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  • 135
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    Cytochromes P450 of the sophorose lipid-producing yeast Candida apicola: Heterogeneity and polymerase chain reaction-mediated cloning of two genes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 565-575 
    Details
    ISSN: 0749-503X
    Keywords: Biosurfactant ; glycolipid ; cytochrome P450 ; Candida apicola ; alkane assimilation ; fatty acid hydroxylation ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Candida apicola belongs to a group of yeasts producing high amounts of surface-active extracellular glycolipids consisting of sophorose and long-chain-ω- and (ω-1)-hydroxy fatty acids. The involvement of cytochrome P450 in the synthesis of sophorose lipid by the hydroxylation of long-chain fatty acids was suggested from a simultaneous increase of cellular P450 content. Hydroxylation studies indicated the existence of multiple P450 forms capable of hydroxylating not only long-chain fatty acids, but also n-alkanes.In this report, two different P450 DNA fragments amplified in a polymerase chain reaction with heterologous primers and chromosomal DNA of Candida apicola were used as homologous probes for the isolation of full-length clones from a genomic library. The open reading frames of both genes encode proteins of 519 amino acids with calculated molecular weights of 58,656 and 58,631, respectively, that contain N-terminal membrane anchor sequences and hallmark residues, in common with other eukaryotic P450s. The deduced amino acid sequences of the C. apicola P450 genes are 84·4% identical. They share 34·5 to 44·1% identity with the proteins of the yeast family CYP52 and about 25% identity with fatty acid hydroxylases of higher eukaryotes (family CYP4A) and of Bacillus megaterium (CYP102). Southern hybridization experiments revealed the existence of further P450-related genes in C. apicola. According to the P450 nomenclature system, the cloned genes were named CYP52E1 and CYP52E2, establishing a new subfamily in yeast family CYP52. The sequences were deposited in the EMBL/GenBank Library under the Accession Numbers X76225 and X87640.
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    Structural and phylogenetic analysis of the actin gene from the yeast Phaffia rhodozyma (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 641-651 
    Details
    ISSN: 0749-503X
    Keywords: Phaffia rhodozyma ; actin gene ; DNA sequencing ; phylogenetic studies ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The gene coding for actin from Phaffia rhodozyma was cloned and sequenced. The Phaffia actin gene contains four intervening sequences and the predicted protein consists of 375 amino acids. The structural features of the Phaffia actin introns were studied and compared with actin introns from seven fungi and yeasts with ascomycetous and basidiomycetous affinity. It was shown that the architecture of the Phaffia introns most resembles that of the basidiomycete Filobasidiella neoformans (perfect stage of Cryptococcus neoformans), whereas least resemblance occurs with the ascomycetous yeasts. Based on the intron structure, the ascomycetous yeasts can be accommodated in one group in that their splice site sequences are very similar and show less homology with the other fungi investigated, including Phaffia. It was demonstrated that the Phaffia actin introns cannot be spliced in Saccharomyces cerevisiae, which shows that the differences found in intron structure are significant. Alignment of the Phaffia actin gene with the actin sequences from the yeasts and fungi investigated showed a high level of homology both on the DNA level and on the protein level. Based on these alignments Phaffia showed highest homology with F. neoformans and both organisms were accommodated in the same cluster. In addition, the actin gene comparisons also supported the distant relationship of Phaffia with the ascomycetous yeasts. These results supported the usefulness of actin sequences for phylogenetic studies. The sequence presented here has been submitted to the EMBL data library under Accession Number X89898.
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    A simplified method for the repeated replacement of yeast chromosomal sequences with in vitro mutations (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 667-672 
    Details
    ISSN: 0749-503X
    Keywords: CYH2S ; counterselection ; PCR ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A strategy for gene replacement in Saccharomyces cerevisiae has been modified to facilitate the repeated substitution of a chromosomal locus with in vitro generated variant sequences, so that the resulting locus contains only the desired mutation and is free of extraneous vector DNA. The construction of an internally deleted chromosomal target locus carrying the counterselectable CYH2S marker and a second positively selectable marker has been simplified; the design of the locus has been altered to increase the frequency of authentic gene replacements obtained upon the subsequent integration of in vitro mutated DNA. The modified chromosomal target locus is amenable to replacement using either of two transformation protocols: (i) integration of a second positively selectable plasmid carrying mutant sequences to form a tandem intermediate structure at the locus; upon counterselection on cycloheximide, all vector sequence is excised to give the desired replacement at high frequency (〉70%); (ii) single-step integration of a linear segment of mutated genomic DNA by selection for cycloheximide resistance. A subsequent screen for the loss of the positively selectable target locus marker detects the desired replacement at modest frequency (〉2%). Polymerase chain reaction using multiple primers in a single amplification reaction is useful for monitoring these variously modified chromosomal loci.
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    Functional characterization of the repeated UASINO element in the promoters of the INO1 and CHO2 genes of yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 653-665 
    Details
    ISSN: 0749-503X
    Keywords: inositol ; choline ; transcription regulation ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In yeast, INO1 and CHO2 gene expression is subject to repression in response to inositol and choline supplementation. The response by both genes to inositol is controlled by a single set of regulatory factors and the highly conserved and repeated UASINO element (consensus: 5′ CATGTGAAAT 3′) that is found in multiple copies in both promoters. However, none of the native elements found in the INO1 and CHO2 promoters constitutes an exact match to the consensus element and the functionality of individual elements from these two promoters has not been tested. In this study, the function of individual putative UASINO elements from both promoters was tested by placing promoter fragments into a reporter construct which lacked a UAS element but contained the TATA element and start of transcription from the yeast CYC1 gene fused to the Escherichia coli lacZ gene. In addition, a set of oligonucleotides containing the consensus UASINO element with the first position systematically modified was also tested for UASINO function. These studies indicated that elements that contain a C or an A as the first base at the 5′ end are functional to varying degrees. The majority of potential UASINO elements from the INO1 promoter were found to be inactive, whereas all of the elements from the CHO2 promoter tested were active. These results are discussed in light of the differential regulation of the two promoters.
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    Sequence analysis of the CEN12 region of Saccharomyces cerevisiae on a 43·7 kb fragment of chromosome XII including an open reading frame homologous to the human cystic fibrosis transmembrane conductance regulator protein CFTR (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 693-708 
    Details
    ISSN: 0749-503X
    Keywords: CEN12 ; DNM1 ; MMM1 ; DRS1 ; SOF1 ; SCD25 ; DPS1/ATS/APSG ; TGL1 ; hMRP1 ; hCFTR ; yeast ; cystic fibrosis ; multidrug resistance ; Pumilio ; MPT5 ; HTR1 ; YGL3 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the framework of the European Union BIOTECH project for systematically sequencing the Saccharomyces cerevisiae genome, we determined the nucleotide sequence of a 43·7 kb DNA fragment spanning the centromeric region of chromosome XII. A novel approach was the distribution of sublibraries prepared by the DNA coordinator (J. Hoheisel, Heidelberg, FRG), using a new hybridization-based DNA mapping method, in order to facilitate ordered sequencing. The sequence contains 22 open reading frames (ORFs) longer than 299 bp, including the published sequences for ATS/DPS1, SCD25, SOF1, DRS1, MMM1, DNM1 and the centromeric region CEN12. Five putative ORF products show similarity to known proteins: the leucine zipper-containing ABC transporter L1313p to the yeast Ycf1p metal resistance protein, to the yeast putative ATP-dependent permease Yhd5p, to the yeast putative proteins Yk83p and Yk84p, to the human cystic fibrosis transmembrane conductance regulator protein (hCFTR) and to the human multidrug resistance-associated protein hMRP1; L1325p to the Drosophila melanogaster Pumilio protein, to the putative yeast regulatory protein Ygl3p and to the yeast protein Mpt5p/Htr1p; L1329p to human lipase A and gastric lipase, to rat lingual lipase and to the putative yeast triglyceride lipase Tgl1p; L1341p to the putative yeast protein Yhg4p; and the leucine zipper-containing L1361p to the two yeast proteins 00953p and Ym8156.08p and to the Arabidopsis thaliana protein HYP1. Eight ORFs show no homology to known sequences in the database, three small ORFs are internal and complementary to larger ones and L1301 is complementary overlapping the ATS/DPS1 gene. Additionally three equally spaced ARS consensus sequences were found. The nucleotide sequence reported here has been submitted to the EMBL data library under the accession number X91488.
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    HST1, a new member of the SIR2 family of genes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 631-640 
    Details
    ISSN: 0749-503X
    Keywords: silencing ; mating type ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A novel Saccharomyces cerevisiae gene, HST1, was identified from among anonymous cDNAs and the complete corresponding genomic clone was isolated and sequenced. HST1 is very closely related to SIR2, showing 71% sequence identity over 84% of its length. Polymerase chain reaction with degenerate primers on S. cerevisiae DNA identified three additional SIR2-related genes designated HST2, HST3 and HST4. The sequences of HST2, HST3 and HST4 correspond to sequences previously released by the S. cerevisiae genome sequencing project as U33335, NCBI gi:965078; X87331, NCBI gi:829135; and Z48784, YD9346.03, respectively. Disruption of HST1 has shown no phenotype with respect to mechanisms in which SIR2 has a role, namely, regional silencing of HMLα, or in rDNA recombination. The sequence of HST1 has been deposited in the DDBJ, EMBL and GenBank at NCBI database under Accession Number L47120.
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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.320120701
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    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 715-722 
    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.320120702
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  • 143
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    Review: To bud until death: The genetics of ageing in the yeast, Saccharomyces (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 623-630 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Individual cells of the budding yeast, Saccharomyces cerevisiae, have a limited division capacity and undergo characteristic changes as they senesce, primarily increasing both their cell size and cell cycle time. The mortality curve for ageing yeast cells can be described by the Gompertz equation, the classical definition for an ageing population. Recent work from several laboratories has demonstrated that genes can determine the yeast lifespan. Studies with the UTH genes have implicated changes in transcriptional silencing during yeast ageing, but the roles of the RAS2, LAG1 and PHB1 genes in regulating yeast longevity are still unclear. What is becoming clearer, however, is that yeast ageing is more than just a bud scar phenomenon.
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    Isolation and characterization of a Δ-9 fatty acid desaturase gene from the oleaginous yeast Cryptococcus curvatus CBS 570 (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 723-730 
    Details
    ISSN: 0749-503X
    Keywords: Δ-9 fatty acid desaturase ; cryptococcus curvatus ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The oleaginous yeast Cryptococcus curvatus is of industrial interest because it can accumulate triacylglycerols up to 60% of the cell dry weight. We are aiming at genetic modification of fatty acid biosynthesis for the production of tailor-made triacylglycerols in C. curvatus. As a first step in the development of a transformation and expression system a gene encoding the Δ-9 fatty acid desaturase of C. curvatus (CBS 570) was cloned. The 1470 bp gene encodes a protein of 493 amino acids with a calculated molecular mass of 55 kDa. The gene shows strong similarity to previous cloned Δ-9 desaturase genes from rat and Saccharomyces cerevisiae, 62 and 72%, respectively. Expression of the Δ-9 desaturase gene was studied. Supplementation of the growth medium with oleic acid (C18:1(c9)) showed a strong repression (90%) on the mRNA level, while supplementation with petroselinic acid (C18:1(c6)) had no effect on the amount of mRNA.
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    A Candida albicans gene encoding a DNA ligase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 893-898 
    Details
    ISSN: 0749-503X
    Keywords: Candida albicans ; IME1 ; CDC9 ; IME2 ; ATP-dependent DNA ligase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A DNA ligase-encoding gene (Ca CDC9) was cloned from Candida albicans by complementation of an ime-1 mutation in Saccharomyces cerevisiae. In this system, IME1 function was assayed using a S. cerevisiae strain with a ime2-promoter-lacZ gene fusion such that following transformation with a C. albicans genomic library, the presence of positive clones was indicated upon the addition of X-gal to sporulation media. Transforming fragments were subcloned in pGEM7 and sequenced. Sequence homology with several ATP-dependent DNA ligases from viruses, fission yeast, human, baker yeast and bacteria was observed. The sequence has been deposited in the EMBL data bank under the Accession Number X95001.
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    Sequence analysis of a 14·6 kb DNA fragment of Saccharomyces cerevisiae chromosome VII reveals SEC27, SSM1b, a putative S-adenosylmethionine-dependent enzyme and six new open reading frames (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 887-892 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VII ; SEC27 ; SSM1b ; S-adenosylmethionine-dependent enzyme ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a fragment from the left arm of Saccharomyces cerevisiae chromosome VII has been determined. Analysis of the 14,607 bp DNA segment reveals nine open reading frames (ORFs) longer than 300 bp. G2827 is the SEC 7 gene, an essential coatomer complex subunit. G2834 encodes SSM1b, a ribosomal protein. The G2838 product shows homology to hypothetical yeast proteins, YIF0 and YE09, of unknown function. The G2830 product shows homology with the cell division protein FtsJ from Escherichia coli, with two hypothetical proteins from yeast, YCF4 and YBR1, and with R74.7, a hypothetical protein from Caenorhabditis elegans. Two of the ORFs are completely internal to longer ones and a third is partially embedded in G2850. The remaining ORFs give no significant homology with proteins in the databases. The sequence has been deposited at the EMBL database under Accession Number X92670.
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    Enrichment of yeast protein tyrosine kinase activity by substrate affinity chromatography (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 833-838 
    Details
    ISSN: 0749-503X
    Keywords: Protein tyrosine kinases ; protein purification ; affinity chromatography ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The direct biochemical analysis of protein tyrosine kinases from yeast has been difficult due to their very low activity in crude cell lysates. Here we present a procedure for the enrichment and partial purification of protein tyrosine kinases from Saccharomyces cerevisiae based on single-step substrate affinity chromatography using a synthetic random co-polymer of glutamic acid and tyrosine. Fractionation of cell lysates on a poly-glutamic acid: tyrosine (4 : 1)-Sepharose affinity column resulted in a 4000-fold increase in tyrosine kinase activity. Active fractions contain at least six potential protein kinases as judged by in situ phosphorylation assay and Western blot analysis using anti-phosphotyrosine. We propose that this protocol may also be useful for the initial identification and purification of tyrosine kinases from other organisms exhibiting low levels of this enzymatic activity in cell lysates.
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  • 148
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    Identification and characterization of cytosolic Hansenula polymorpha proteins belonging to the Hsp70 protein family (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 849-857 
    Details
    ISSN: 0749-503X
    Keywords: heat shock proteins ; molecular chaperones ; stress tolerance ; peroxisome biogenesis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated two members of the Hsp70 protein family from the yeast Hansenula polymorpha using affinity chromatography. Both proteins were located in the cytoplasm. One of these, designated Hsp72, was inducible in nature (e.g. by heat shock). The second protein (designated Hsc74) was constitutively present. Peptides derived from both Hsp72 and Hsc74 showed sequence homology to the cytosolic Saccharomyces cerevisiae Hsp70s, Ssa1p and Ssa2p. The gene encoding Hsp72 (designated HSA1) was cloned, sequenced and used to construct HSA1 disruption and HSA1 overexpression strains. Comparison of the stress tolerances of these strains with those of wild-type H. polymorpha revealed that HSA1 overexpression negatively affected the tolerance of the cells to killing effects of temperature or ethanol, but enhanced the tolerance to copper and cadmium. The tolerance for other chemicals (arsenite, arsenate, H2O2) or to high osmolarity was unaffected by either deletion or overexpression of HSA1. The nucleotide sequence of HSA1 was submitted to the EMBL data library and given the Accession Number Z29379.
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    Review: Subcellular traffic of the plasma membrane H+-ATPase in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 907-916 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 2 Ill.
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    Characterization of cwl1+, a gene from Schizosaccharomyces pombe whose overexpression causes cell lysis (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 983-990 
    Details
    ISSN: 0749-503X
    Keywords: fission yeast ; dominant genetics ; cell wall regulation ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: From a Schizosaccharomyces pombe genomic library we have isolated the gene cwl1+ that causes cell lysis when it is overexpressed in the absence of an osmotic stabilizer. Southern hybridization showed that cwl1+ exists as a single copy in the S. pombe genome. The cwl1+ gene nucleotide sequence revealed a putative open reading frame of 924 bp encoding a polypeptide of 308 amino acids with a calculated Mr of 27 000. The cwl1+ DNA hybridizes to a major RNA transcript of 1·5 kb whose 5′ end maps at a position 452 bp upstream from the predicted translation start. Comparison of the amino acid sequence with those included in the current databases, showed no significant similarity to any known sequences. Cells overexpressing the cwl1+ gene under the control of the S. pombe nmt inducible promoter displayed a reduced cell wall content, were unable to separate after division and lysed drastically in the absence of osmotic stabilizer. Disruption of the cwl1+ gene caused no noticeable phenotype. The sequence has been deposited in the EMBL data library under Accession Number X9445.
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    A useful colony colour phenotype associated with the yeast selectable/counter-selectable marker MET15 (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 939-941 
    Details
    ISSN: 0749-503X
    Keywords: methionine ; sectored colonies ; colour marker ; MET15 ; Saccharomyces cerevisiae ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Strains of Saccharomyces cerevisiae bearing null alleles of the met15 gene are methionine auxotrophs and become darkly pigmented in the presence of Pb2+ ions (Ono et al. (1991). Appl. Env. Microbiol. 57, 3183-3186). We describe the cloning of a useful fragment of the MET15 locus which complements both the methionine requirement and the colony colour phenotype. This colony colour phenotype is very useful for genetic screens and may be applicable for use in other yeast species. The combination of the size of MET15, along with its counter-selectability and the colour of met15 mutations make this perhaps the most versatile yeast genetic marker.
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    Sequence of the GLT1 gene from Saccharomyces cerevisiae reveals the domain structure of yeast glutamate synthase (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1359-1366 
    Details
    ISSN: 0749-503X
    Keywords: glutamate synthase ; S. cerevisiae ; chromosome IV ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Glutamate synthase (GOGAT) and glutamine synthetase play a crucial role in ammonium assimilation and glutamate biosynthesis in the yeast Saccharomyces cerevisiae. The GOGAT enzyme has been purified and the GOGAT structural gene (GLT1) has been cloned, showing that this enzyme is a homotrimeric protein with a monomeric size of 199kDa.We report the GLT1 nucleotide sequence and the amino acid sequence of its deduced protein product. Our results show that there is a high conservation with the corresponding genes of Escherichia coli, Medicago sativa (alfalfa) and Zea mais (maize). Binding domains for glutamine, cofactors (FMN and NADH) and the cysteine clusters (which comprise the iron-sulfur centres) were tentatively identified on the basis of sequence comparison with GOGAT sequences from E. coli, alfalfa and maize. The sequence of GLT1 has been deposited in the EMBL data library under Accession Number X89221.
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    Molecular genetics of ICL2, encoding a non-functional isocitrate lyase in saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1285-1295 
    Details
    ISSN: 0749-503X
    Keywords: ICL2 ; glyoxylate cycle ; deletion analysis ; transcription ; gluconeogenesis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this work, we identified an open reading frame 5′ to the yeast HALI gene, that shares a 38% identity in the deduced amino acid sequence with gluconeogenic enzyme isocitrate lyase, encoded by ICL1. We therefore termed the new gene ICL2. The latter is not capable of complementing an icl1 deletion for growth on ethanol neither in its original context, nor when expressed under the control of the glycolytic PFK2 promoter. Nevertheless, fusions of the 5′-non-coding region of ICL2 to the lacZ reporter gene revealed that the gene is transcribed and that the transcriptional regulation is similar to that of other gluconeogenic genes, i.e. high-level expression on ethanol that is drastically reduced on glucose media. Therefore, we attribute the lack of complementation to a lack of function of the encoded protein as an isocitrate lyase. The deduced amino acid sequences of Icl1 and Icl2 differ in a conserved motif used to identify isocitrate lyases, the hexapeptide KKCGHM, where the second lysine residue of Icl1 is replaced by an arginine in Icl2. However, we here demonstrated by in vitro mutagenesis of ICL1 that such an exchange, even though it affects Icl activity to some degree, does not lead to a complete lack of function. Thus, the results presented in this work argue for ICL2 encoding a non-functional isocitrate lyase and provide evidence that lysine 216 of Icl1 is not essential for catalysis. This sequence is deposited as accession number Z48951 entered on4 April 1995 by Barrel et al.
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    Evidence for a selective and electroneutral K+/H+-exchange in Saccharomyces cerevisiae using plasma membrane vesicles (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1301-1313 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; plasma membrane purification ; vesicles reconstitution ; K+/H+-exchange ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The existence of a K+/H+ transport system in plasma membrane vesicles from Saccharomyces cerevisiae is demonstrated using fluorimetric monitoring of proton fluxes across vesicles (ACMA fluorescence quenching). Plasma membrane vesicles used for this study were obtained by a purification/reconstitution protocol based on differential and discontinuous sucrose gradient centrifugations followed by an octylglucoside dilution/gel filtration procedure. This method produces a high percentage of tightly-sealed inside-out plasma membrane vesicles. In these vesicles, the K+/H+ transport system, which is able to catalyse both K+ influx and efflux, is mainly driven by the K+ transmembrane gradient and can function even if the plasma membrane H+-ATPase is not active. Using the anionic oxonol VI and the cationic DISC2(5) probes, it was shown that a membrane potential is not created during K+ fluxes. Such a dye response argues for the presence of a K+/H+ exchange system in S. cerevisiae plasma membrane and established the non-electrogenic character of the transport. The maximal rate of exchange is obtained at pH 6·8. This reversible transport system presents a high selectivity for K+ among other monovalent cations and a higher affinity for the K+ influx into the vesicles (exit from cells). The possible role of this K+/H+ exchange system in regulation of internal potassium concentration in S. cerevisiae is discussed.
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    Identification of a gene encoding a homocitrate synthase isoenzyme of Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1315-1320 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; lysine ; homocitrate synthase ; nifV gene ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In Saccharomyces cerevisiae, most of the LYS structural genes have been identified except the genes encoding homocitrate synthase and α-aminoadipate aminotransferase. Expression of several LYS genes responds to an induction mechanism mediated by the product of LYS14 and an intermediate of the pathway, α-aminoadipate semialdehyde (αAASA) as an inducer. This activation is modulated by the presence of lysine in the growth medium leading to an apparent repression. Since the first enzyme of the pathway, homocitrate synthase, is feedback inhibited by lysine, it could be a major element in the control of αAASA supply.During the sequencing of chromosome IV of S. cerevisiae, the sequence of ORF D1298 showing a significant similarity with the nifV gene of Azotobacter vinelandii was reported. Disruption and overexpression of ORF D1298 demonstrate that this gene, named LYS20, encodes a homocitrate synthase. The disrupted segregants are able to grow on minimal medium and exhibit reduced but significant homocitrate synthase indicating that this activity is catalysed by at least two isoenzymes. We have also shown that the product of LYS20 is responsible for the greater part of the lysine production.The different isoforms are sensitive to inhibition by lysine but only the expression of LYS20 is strongly repressed by lysine. The N-terminal end of homocitrate synthase isoform coded by LYS20 contains no typical mitochondrial targeting sequence, suggesting that this enzyme is not located in the mitochondria.
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    Intergenic flip flop, a method for systematic gene disruption and cloning in yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1351-1357 
    Details
    ISSN: 0749-503X
    Keywords: gene deletion ; gap repair ; polymerase chain reaction ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have developed a strategy named Intergenic Flip Flop which, for each gene, allows us to produce in one experiment both a disrupting cassette and a plasmid for gap repair. The same method can also be used to insert a reporter gene downstream from the promoter. This approach extends the polymerase chain reaction (PCR)-based strategy proposed by Maftahi et al. 1996. Our method consists of PCR amplification of the two flanking intergenic regions of the open reading frame (ORF) of interest, using two sets of oligonucleotides. Each PCR product is flanked by two short defined nucleotidic sequences with a unique restriction site, allowing subsequent hybridization between them. The association of the two amplimers by the complementary sequences either in the same orientation as in genomic DNA or in the opposite orientation, allows the generation, after PCR, of two distinct cassettes which can be cloned into suitable vectors. When the amplimer in the head-to-tail orientation is cloned in a vector containing a selective marker for yeast such as G418 resistance, it provides a disrupting cassette after cleavage at the unique restriction site introduced by the PCR between the two intergenic amplimers. The amplimer with a direct orientation cloned into a yeast vector, after cleavage at the unique restriction site between the intergenic regions, permits cloning by gap repair of the gene of interest in yeast. Finally, a reporter gene can be inserted in the same plasmid. We report here the successful application of this strategy to an ORF of chromosome XIV of Saccharomyces cerevisiae: N1216.
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    CtCdc55p and CtHal3p: Two putative regulatory proteins from Candida tropicalis with long acidic domains (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1321-1329 
    Details
    ISSN: 0749-503X
    Keywords: Candida tropicalis ; CDC55 ; HAL3 ; protein phosphatase ; acidic domain ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The salt-tolerance gene HAL3 from Saccharomyces cerevisiae encodes a novel regulatory protein (Hal3p) which modulates the expression of the ENA1 sodium-extrusion ATPase (Ferrando et al., Mol. Cell. Biol. vol.15, 1995, pp.5470-5481). Hal3p contains an essential acidic domain rich in aspartates at its carboxyl terminus. We have isolated two cross-hybridizing genes from a genomic library of Candida tropicalis. One of the genes (CtHAL3) is a true homolog of HAL3 and it partially complements the salt sensitivity of a S. cerevisiae hal3 mutant. The activity of CtHAL3 was equivalent to that of an open reading frame (YKL088w) identified by genome sequencing of S. cerevisiae and with homology to HAL3. The other cross-hybridizing gene (CtCDC55) is a CDC55 homolog, encoding a protein with an internal acidic domain not present in the S. cerevisiae CDC55 product. Cdc55p is a regulatory subunit of protein phosphatase 2A and CtCDC55 complements the cold sensitivity of a S. cerevisiae cdc55 mutant. The presence of acidic domains in different putative regulatory proteins may suggest a role for this type of domain in molecular interactions. Sequences have been deposited in the EMBL data library under Accession Numbers X88899 (CtCDC55) and X88900 (CtHAL3).
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    Reduced pyruvate decarboxylase and increased glycerol-3-phosphate dehydrogenase [NAD+] levels enhance glycerol production in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1331-1337 
    Details
    ISSN: 0749-503X
    Keywords: pyruvate decarboxylase ; glycerol-3-phosphate dehydrogenase ; glycerol production ; Saccharomyces cerevisiae ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This investigation deals with factors affecting the production of glycerol in Saccharomyces cerevisiae. In particular, the impact of reduced pyruvate-decarboxylase (PDC) and increased NAD-dependent glycerol-3-phosphate dehydrogenase (GPD) levels was studied. The glycerol yield was 4·7 times (a pdc mutant exhibiting 19% of normal PDC activity) and 6·5 times (a strain exhibiting 20-fold increased GPD activity resulting from overexpression of GPD1 gene) that of the wild type. In the strain carrying both enzyme activity alterations, the glycerol yield was 8·1 times higher than that of the wild type. In all cases, the substantial increase in glycerol yield was associated with a reduction in ethanol yield and a higher by-product formation.The rate of glycerol formation in the pdc mutant was, due to a slower rate of glucose catabolism, only twice that of the wild type, and was increased by GPD1 overexpression to three times that of the wild-type level. Overexpression of GPD1 in the wild-type background, however, led to a six- to seven-fold increase in the rate of glycerol formation. The experimental work clearly demonstrates the rate-limiting role of GPD in glycerol formation in S. cerevisiae.
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    Mutational analysis of the gene for Schizosaccharomyces pombe RNase MRP RNA, mrp1, using plasmid shuffle by counterselection on canavanine (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1393-1405 
    Details
    ISSN: 0749-503X
    Keywords: Plasmid shuffle ; RNase MRP RNA ; Schizosaccharomyces pombe ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Reverse genetics in fission yeast is hindered by the lack of a versatile established plasmid shuffle system. In order to screen efficiently and accurately through plasmid-borne mutations in the essential gene for the RNA component of RNase MRP, mrp1, we have developed a system for plasmid shuffling in fission yeast using counterselection on canavanine. The system takes advantage of the ability of the Saccharomyces cerevisiae CAN1 gene to complement a Schizosaccharomyces pombe can1-1 mutation. Two general use plasmids were constructed that allow directional cloning and initial selection for histidine before counterselection by canavanine. The strain constructed for plasmid shuffling carries auxotrophic markers for ade6, leu1, ura4 and his3 along with the can1-1 mutation. Using this system we examined several partial deletions and point mutations in conserved nucleotides of Schizosaccharomyces pombe RNase MRP RNA for their ability to complement a chromosomal deletion of the mrp1 gene. The degree of background canavanine resistance as well as plasmid-plasmid recombination encountered in these experiments was sufficiently low to suggest that the system we have set up for counterselection by canavanine in fission yeast using multicopy plasmids will be widely useful.
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    Cloning and sequencing of the LYS1 gene encoding homocitrate synthase in the yeast Yarrowia lipolytica (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1459-1469 
    Details
    ISSN: 0749-503X
    Keywords: Yarrowia lipolytica ; homocitrate synthase ; α-isopropyl malate synthase ; mitochondria ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The α-aminoadipate pathway for the biosynthesis of lysine is present only in fungi and euglena. The first step in the pathway is the condensation of acetyl-CoA and α-ketoglutarate into homocitrate, and this step is carried out by the enzyme homocitrate synthase (EC 4.1.3.21). In spite of extensive genetic analysis, no mutation affecting this step has been isolated until now in model organisms such as Saccharomyces cerevisiae or Neurospora crassa, although identification of mutations affecting the structural gene (LYS1) for homocitrate synthase was reported in the yeast Yarrowia lipolytica several years ago. Here we used these mutants for the cloning and sequencing of the Yarrowia LYS1 gene. The LYS1 gene encodes a predicted 446 amino acid polypeptide, with a molecular mass of 48 442 Da. The Lys1p sequence displays two regions, one near the N-terminal section and the other in the central region, that contain conserved signatures found in prokaryotic homocitrate synthases (nifV genes of Azotobacter vinelandii and Klebsiella pneumoniae), as well as in all α-isopropyl malate synthases so far described. A putative mitochondrial targeting signal of 41-45 amino acids is predicted at the N-terminus. The Lys1p sequence shows 84% identity at the amino acid level with the putative product of open reading frame D1298 of S. cerevisiae. Northern blot hybridizations revealed a LYS1 transcript of approximately 1·7 kb in Y. lipolytica. Deletion of the LYS1 gene resulted in a Lys- phenotype. Our results indicate that we cloned the structural gene for homocitrate synthase in Y. lipolytica, and that the enzyme is encoded by a single gene in this yeast. The sequence presented here has been submitted to the EMBL data library under Accession Number Z49114.
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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.320121401
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    HAM1, the gene controlling 6-N-hydroxylaminopurine sensitivity and mutagenesis in the yeast Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 17-29 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; 6-N-hydroxylaminopurine ; hypermutability ; molecular cloning and sequencing ; purine metabolism ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ham1 mutant of yeast Saccharomyces cerevisiae is sensitive to the mutagenic and lethal effects of the base analog, 6-N-hydroxylaminopurine (HAP). We have isolated a clone from a centromere-plasmid-based genomic library complementing HAP sensitivity of the ham1 strain. After subcloning, a 3·4 kb functional fragment was sequenced. It contained three open reading frames (ORFs) corresponding to proteins 353, 197 and 184 amino acids long. LEU2+ disruptions of the promoter and N-terminal part of the gene coding 197 amino acids long protein led to moderate and strong sensitivity to HAP, respectively, and were allelic to the original ham1-1 mutation. Thus this ORF represents the HAM1 gene. The deduced amino acid sequence of HAM1 protein was not similar to any protein sequence of the SwissProt database. The HAM1 gene was localized on the right arm of chromosome X between cdc8 and cdc11. Spontaneous mutagenesis was not affected by the ham1::LEU2 disruption mutation.
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    Analysis of a 26 kb region on the left arm of yeast chromosome XV (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 67-76 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; ORFs ; predictable functions ; regulatory elements ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the framework of the EC programme for sequencing yeast chromosome XV, we have determined the nucleotide sequence of a 26 kb region. Subsequent analysis revealed 13 non-overlapping open reading frames, three of which correspond to known yeast genes. A pair of tRNA genes associated with remnant Ty elements were localized in this region. From structural parameters and/or similarity searches with entries in the current data libraries, a preliminary functional assessment of several of the putative novel gene products can be made. The gene density in this region amounts to one gene in 2 kb. Protein coding regions occupy 61% of the total DNA sequence. Within the intergenic regions, potential regulatory elements can be predicted. The data obtained here may serve as a basis for a more detailed biochemical analysis of the novel genes. The complete nucleotide sequence of the 26 kb segment as depicted in Figure 1 has been deposited at the EBI data library under Accession Number X91067.
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    Cloning and characterization of the Pichia pastoris PRC1 gene encoding carboxypeptidase Y (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 31-40 
    Details
    ISSN: 0749-503X
    Keywords: Pichia pastoris ; carboxypeptidase Y ; PRC1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We purified a 58 kDa serine protease from culture-supernatant of Pichia pastoris and found that the NH2-terminal amino acid sequence of this protease is closely homologous to that of mature protein of Saccharomyces cerevisiae carboxypeptidase Y (CPY), which is encoded by the PRC1 gene. Using the S. cerevisiae PRC1 gene as a hybridization probe, a cross-hybridizing fragment of P. pastoris genomic DNA was identified and the gene, PRC1, encoding CPY, was cloned. The open reading frame of the P. pastoris PRC1 gene consists of 1569 bp encoding a protein of 523 amino acids. The molecular mass of the protein is calculated to be 59·44 kDa without sugar chains. The protein comprises 20 amino acids of pre (signal)-peptide, 87 amino acids of pro-peptide and 416 amino acids of mature peptide, and has four N-glycosylation sites. The NH2-terminal amino acid sequence of mature peptide is completely identical with that of the protease purified from the culture-supernatant. There is 61% identity between the amino acid sequences of P. pastoris Prc1p and S. cerevisiae Prc1p. Chromosomal disruption of the PRC1 gene resulted in the loss of CPY activity. Over-expression of the PRC1 gene under regulation of the P. pastoris AOX1 promoter resulted in accumulation of a large amount of active CPY in the intracellular fraction, and secretion of a slightly larger molecule that is thought to be pro-CPY. The nucleotide sequence data reported in this paper will appear in the EMBL Nucleotide Sequence Databases under the Accession Number X87987.
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    Separate roles for N- and C-termini of the STE4 (β) subunit of the Saccharomyces cerevisiae G protein in the mediation of the growth arrest. Lack of growth-arresting activity of mammalian βγ complexes (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 41-51 
    Details
    ISSN: 0749-503X
    Keywords: STE4 ; STE18 ; G proteins ; signal transduction ; yeast ; structure-function ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mating pheromone signal transduction in Saccharomyces cerevisiae involves a G protein composed to Scg1p (Gpa1p), Ste4p and Ste18p subunits, homologous to the α, β and γ subunits of mammalian G proteins. Growth arrest in G1 phase is activated by the Ste4p/Ste18p complex via a downstream pathway and it is negatively controlled by the Scg1p subunit. Here we explored whether mammalian β or γ subunits could functionally substitute for their yeast homologues. While no evidence was obtained for functional replacement of Ste4p and Ste18p, we found that overexpression of Ste18p potentiated the effect of hybrid proteins in which the N terminus of the Ste4p subunit was replaced by that of the mammalian β, ste4 mutants having deletions in the N terminus showed a decreased activity in signalling to the downstream effector of the pheromone response. This defect was totally cured by overexpression of Ste18p, indicating that the truncated forms of Ste4p have retained their ability to form an active complex with Ste18p. Removal of six amino acids from the C terminus of Ste4p rendered a completely inactive subunit and this defect persisted in hybrids where the C terminus was placed by that of the β subunit, indicating that the C terminus of Ste4p is essential to trigger the effector of the yeast pheromone response pathway.
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  • 166
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    Erratum to: Sequence, map position and genome organization of the RPL 17B gene, encoding ribosomal protein L17b in Saccharomyces cerevisiae (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 91-91 
    Details
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 167
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    Sequencing of a 23 kb fragment from Saccharomyces cerevisiae chromosome VI (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 77-84 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VI ; SEC4 ; MSH4 ; TYA ; TYB ; SPB4 ; DEG1 ; NIC96 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Plasmid clone gapB and lambda phage clone 4682, which contain fragments of Saccharomyces cerevisiae chromosome VI, were analysed. A 23 kb sequence was determined and ten open reading frames (ORFs) were revealed. Among them, five ORFs were identical to five yeast genes (SEC4, MSH4, SPB4, DEG1 and NIC96), two were identical to transposable elements (TYA and TYB), one (gapBorfF003) was highly homologous to a yeast expressed sequence tag, and another (4682orfF002) was predicted to be a nuclear protein. Sequence data have been submitted to DDBJ/EMBL/GenBank data library under Accession Number D44604 (clone gapB) and D44600 (clone 4682), respectively.
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  • 168
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    Nucleotide sequence analysis of a 32,500 bp region of the right arm of Saccharomyces cerevisiae chromosome IV (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 85-90 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; genome sequencing ; chromosome IV ; DOA4 ; UBC5 ; UBC3 ; DNA52 ; ribosomal protein gene ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a region containing 32.5 kb of the right arm of chromosome IV of Saccharomyces cerevisiae. Twenty open reading frames (ORFs) greater than 100 amino acids could be identified in this region. Six ORFs correspond to known yeast genes, including DOA4, UBC5 and UBC3, the gene products of which are involved in ubiquitin metabolism. UBC5 is preceded by the two tRNA genes tRNA-Arg2 and tRNA-Asp. Six genes were discovered with homologies to non-yeast genes or with homologies to other yeast ORFs. One of these could be identified as ribosomal protein gene RPS13. The putative function of eight ORFs remains unclear because comparison to different DNA or protein databases revealed no significant patterns. The sequence from cosmid 2F21 was obtained entirely by a combined subcloning and walking primer strategy, and has been deposited in the EMBL data library under Accession Number X84162.
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  • 169
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    The yeast growth control gene GRC5 is highly homologous to the mammalian putative tumor suppressor gene QM (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 53-65 
    Details
    ISSN: 0749-503X
    Keywords: growth control ; cytoskeleton ; QM ; S. cerevisiae ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We isolated the Saccharomyces cerevisiae GRC5 (growth control) gene by functional complementation in vivo of a ts (temperature sensitive) mutation. Phenotypic analysis suggested involvement of GRC5 in cell growth and proliferation. Mutant cells arrest their cell cycles after one to three cell divisions predominantly as mother cells with a large bud. In the region of the septum, a massive accumulation of cell wall material is observed. The mother and daughter nuclei are well separated and spindles are no longer present, while the cytoskeleton is of aberrant appearance. Arrested cells do not perform protein synthesis and are unable to mate. Furthermore, grc5-1ts cells rapidly lose viability at the restrictive temperature (37°C) only on full media, but not under nitrogen-starvation conditions, indicating that proper response to this nutrient limitation is still intact in mutant cells after cell cycle arrest. The sequence of GRC5 translates into a basic protein of 221 amino acids with a corresponding Mr of 25·4 kDa. GRC5 is a member of the highly conserved QM gene family, members of which have been reported from plants, invertebrates and vertebrates. The amino acid sequence of GRC5 over its entire length is more than 60% identical to the human QM protein, expression of which is associated with loss of the tumorigenic phenotype in a cell line derived from Wilms' tumor, a malignancy of the embyronic kidney. Here, we show that GRC5 is an essential yeast gene, the function of which as inferred from analysis of the grc5-1ts mutant is crucial for establishment of proper cytoskeletal structure and regulation of growth in yeast cells.
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  • 170
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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.320120101
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  • 171
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    Current awareness on yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 93-100 
    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.320120102
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  • 172
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    Review: Biosynthesis and function of yeast vacuolar proteases (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 1-16 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; lysosome ; sorting ; proteolytic activation ; endocytosis autophagocytosis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The yeast vacuole, which is equivalent to the lysosome of higher eukaryotes, is one of the best characterized degradative organelles. This review describes the biosynthesis and function of yeast vacuolar proteases. Most of these enzymes are delivered to the vacuole via the early compartments of the secretory pathway and the endosome, while one of them is directly imported from the cytoplasm. The proteases are synthesized as precursors which undergo many post-translational modifications before the final active form is generated. Proteolytic activation by removal of propeptides is performed by proteinase A and/or proteinase B in an intricate cascade-like fashion. Recent developments in the analysis of the functions of vacuolar proteolysis are described. Substrates of the vacuolar proteases are mostly imported via endocytosis or autophagocytosis, and vacuolar proteolysis appears to be mainly important under nutritional stress conditions and sporulation.
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  • 173
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    Systematic mapping of autonomously replicating sequences on chromosome V of Saccharomyces cerevisiae using a novel strategy (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 101-113 
    Details
    ISSN: 0749-503X
    Keywords: ARS ; genome analysis ; chromosome V ; physical mapping ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have developed a new procedure for easy and rapid identification of autonomously replicating sequences (ARSs) and have applied it to the analysis of chromosome V of Saccharomyces cerevisiae. The procedure makes use of the ordered λ phage clone bank of this chromosome that we have constructed, and includes transposition of a mini-transposon and selection of transposon-containing derivatives, isolation of their DNA and circularization at their cos-ends, transformation of yeast cells with the circularized DNA, and scoring transformation frequency. The transposon used was derived from Tn5supF, contained the yeast LEU2 gene, and was placed, together with the hyperactive transposase gene, on a mini-F plasmid for stable maintenance in Escherichia coli K-12. Sixteen regions of chromosome V showing ARS activity were identified, of which 12 were newly found in this work. Thus, the procedure will be useful for systematic genomic scale analysis of ARSs in yeast and related organisms in which ordered clone banks have been established. The average distance between adjacent ARS-containing regions was approximately 40 kb. Two-dimensional gel electrophoretic analysis of chromosome replication indicated that one of the newly identified ARSs was functional as an actual in situ replication origin, at least under the conditions employed.
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  • 174
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    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.
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  • 175
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    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.
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  • 176
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    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.
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  • 177
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    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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  • 178
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    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.
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  • 179
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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.320120201
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  • 180
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    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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  • 181
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    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.
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  • 182
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    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.
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  • 183
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    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.
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  • 184
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    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.
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  • 185
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    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.
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    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.
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    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.
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  • 188
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    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.
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    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.
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  • 190
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    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.
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    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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    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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    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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    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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    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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