bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phylogenetic study of ribosomal DNA of cactophilic Pichia species by restriction mapping
Authors: Shen Randy , Lachance Marc‐André 
Published: May 2005

AbstractThe rDNAs of strains of the cactophilic Pichia species P. amethionina, P. antillensis, P. barkeri, P. cactophila, P. caribaea, P. deserticola, P. heedii, P. kluyveri, P. norvegensis, P. opuntiae, P. pseudocactophila, P. thermotolerans and their varieties and anamorphs were mapped with 15 restriction endonucleases, and compared to P. membranaefaciens and P. salictaria as possible non‐cactophilic relatives. The existence of species complexes among those taxa was confirmed. P. membranaefaciens was a plausible ancestral species, and its closest relative in the cactophilic group was P. deserticola. These two species appeared to be moderately related to P. heedii and to P. barkeri, but the latter was shown clearly to belong to the P. kluyveri complex, in spite of a 6 mol% G+C difference in their nuclear DNAs. P. cactophila and P. pseudocactophila ostensibly emerged from P. norvegensis, a facultatively cactophilic yeast. The P. amethionina, P. cactophila and P. opuntiae species complexes appeared independent from one another and from all other species studied. P. salictaria did not appear to be related to P. amethionina.
Sequence of the open reading frame of the FL01 gene from Saccharomyces cerevisiae
Authors: Teunissen A. W. R. H. , Holub E. , Van Der Hucht J. , Van Den Berg J. A. , Steensma H. Y. 
Published: May 2005

AbstractThe cloned part of the flocculation gene FLO1 of Saccharomyces cerevisiae (Teunissen, A.W.R.H., van den Berg, J.A. and Steensma, H.Y. (1993). Physical localization of the flocculation gene FLO1 on chromosome I of Saccharomyces cerevisiae, Yeast, in press) has been sequenced. The sequence contains a large open reading frame of 2685 bp. The amino acid sequence of the putative protein reveals a serine‐ and threonine‐rich C‐terminus (46%), the presence of repeated sequences and a possible secretion signal at the N‐terminus. Although the sequence is not complete (we assume the missing fragment consists of repeat units), these data strongly suggest that the protein is located in the cell wall, and thus may be directly involved in the flocculation process.
A yeast gene encoding a putative RNA helicase of the “DEAD”‐box family
Authors: Strahl‐Bolsinger Sabine , Tanner Widmar 
Published: May 2005

AbstractAn unknown open reading frame from Saccharomyces cerevisiae was identified and sequenced. The predicted amino acid sequence shows high homology to the DEAD‐box family of proteins. Gene disruption revealed that the gene is not essential for yeast but necessary for normal cell growth.
Endomitotic diploidization of Saccharomyces cerevisiae by heat treatment during spore germination
Authors: Tani Yoshinobu , Tomohiro Yukio , Miyata Akira , Furukawa Kensuke , Hayashida Shinsaku 
Published: May 2005

AbstractDiploid cells with ability to mate, hereafter referred to as diploid mater cells, were obtained at significant frequencies by the heat treatment of haploid spores at the early germination stage in Saccharomyces cerevisiae heterothallic strain CG5M (a/α diploid cells heterozygous for five auxotrophic markers). The highest frequency (ca. 11%) of diploidization was obtained from viable cells after heat treatment at 55°C for 10 min when spores were precultivated for 30 min in liquid medium to initiate the germination. The diploid mater cells obtained were homozygous for mating type and for the auxotrophic markers. The diploidization of a spore is thus concluded to be due to endomitotic events in germinating heat‐treated spores.
Expression of HIV‐1 nef in yeast: The 27 kDa nef protein is myristylated and fractionates with the nucleus
Authors: Macreadie Ian G. , Ward Alister C. , Failla Paul , Azad Ahmed A. , Grgacic Elizabeth , McPhee Dale 
Published: May 2005

AbstractThe nef gene of human immunodeficiency virus type 1 (HIV‐1) has been expressed in the yeast Saccharomyces cerevisiae to produce native Nef proteins. The proteins of Mr 27 kDa and 25 kDa, produced by translation from the first and second start codons of the nef gene react with human HIV‐1 antisera. Under low‐level steady‐state expression conditions, Nef27 undergoes myristylation and is targeted to the nuclear fraction while Nef25 is not myristylated and not nuclear localized. When produced rapidly and to high levels, Nef27 is initially present in the cytoplasm as a soluble myristylated protein that later fractionates with the nucleus.
Assay of trehalose with acid trehalase purified from Saccharomyces cerevisiae
Authors: Kienle Iris , Burgert Mark Us , Holzer Helmut 
Published: May 2005

AbstractAn enzymatic end‐point assay of trehalose using acid trehalase from yeast is described. After quantitative hydrolysis of trehalose by acid trehalase, the resulting glucose is assayed with the commercially available glucose oxidase/peroxidase dye system. Pre‐existing glucose is determined in a control reaction from which acid trehalase is omitted. When intact cells are analysed for trehalose, pre‐existing glucose can be washed out with ice‐cold water without reducing the trehalose content of the cells. A convenient method for extraction of trehalose from intact yeast cells is heating for 20 min at 95°C followed by centrifugation. The specificity of the assay is determined by the specificity of the acid trehalase preparation used. As described previously (Mittenbühler, K. and Holzer, H., 1988, J. Biol. Chem. 263, 8537–8543; Mittenbühler, K., 1988, Thesis, University of Freiburg), the following sugars and sugar derivatives do not form glucose when incubated with purified acid trehalase: sucrose, cellobiose, mellobiose, raffinose, maltose, lactose, glucose‐6‐phosphate, glucose‐1‐phosphate, galactose. The application of the new trehalose assay to yeast cells grown to different growth stages and at various temperatures is presented.
RIM2, MSI1 and PGI1 and located within an 8 kb segment of Saccharomyces cerevisiae chromosome II, which also contains the putative ribosomal gene L21 and a new putative essential gene with a leucine zipper motif
Authors: Démolis Nadine , Mallet Laurent , Bussereau Françoise , Jacquet Michel 
Published: May 2005

AbstractWe report the DNA sequence of an 8 kb segment localized on the right arm of chromosome II from Saccharomyces cerevisiae. The sequence reveals the presence of eight open reading frames (ORFs). Three of them, YBR1402, YBR1405 and YBR1406 are previously sequenced genes, respectively the RIM2 (replication in mitochondria), MSI1 (multicopy suppressor of IRA1 gene) and PGI1 (phosphoglucoisomerase) genes. The predicted product of the ORF YBR1401 could be the putative yeast ribosomal protein L21. A new essential gene, YBR1403, has been identified by disruption; it possesses a leucine zipper motif.
TA‐repeat microsatellites are closely associated with ARS consensus sequences in yeast chromosome III
Authors: Valle Giorgio 
Published: May 2005

AbstractMicrosatellites are repeats of very short sequences of DNA, interspersed in the genome. In this paper, the occurrence of the two‐base repeat microsatellites has been investigated in the DNA sequence of yeast chromosome III. Only AT‐repeats were found at a significantly high frequency. Some of the regions with the highest concentration of AT‐repeats were located and further analysed, showing a close association with the core consensus of autonomously replicating sequences.
Linguistic analysis of chromosome III DNA sequence of Saccharomyces cerevisiae
Authors: Kalogeropoulos Angelos 
Published: May 2005

AbstractThe analysis of the Saccharomyces cerevisiae chromosome III DNA sequence by computer (‘in silico’) permits the definition of its linguistic characteristics. These characteristics include the designation of non‐randomly occurring oligonucleotides, their distribution along the chromosome, and the distribution of some particular homopolymers. All these elements may contribute to the understanding of the organization of information on the chromosome.
Cloning and sequence of ADP‐ribosylation factor 1 (ARF1) from Schizosaccharomyces pombe
Authors: Erickson F. Les. , Hannig Ernest M. , Krasinskas Alyssa , Kahn Richard A. 
Published: May 2005

AbstractA gene encoding a homologue of the ADP‐ribosylation factor (ARF) family of small GTP binding proteins was cloned from a Schizosaccharomyces pombe cDNA library by a functional screen of suppressors of sensitivity to 3‐aminotriazole in a gcn3 null strain of Saccharomyces cerevisiae. Two independent isolates each contained the full coding region of the ARF1 gene. The encoded SpARF1 protein has a predicted molecular weight of 20 618 and is 88% and 79% identical to human and S. cerevisiae ARF1 proteins, respectively. As independent isolates were obtained, this effect of the SpARF1 appears to be a real phenomenon, but cannot currently be easily understood within the context of the evidence for a role(s) for ARF proteins in the protein secretory pathway.
Comparison of the biochemical and biological functions of tyrosine phosphatases from fission yeast, budding yeast and animal cells
Authors: Hannig Gerhard , Ottilie Sabine , Schievella Andrea R. , Erikson Raymond L. 
Published: May 2005

AbstractIn a previous communication, we have shown that two protein tyrosine phosphatases (PTPases) from fission yeast, pyp1+ and pyp2+, act as novel inhibitors of mitosis upstream of the wee1+ lmik1+ pathway (Ottilie et al., 1992). Here we describe that both genes possess intrinsic PTPase activity as judged by in vitro PTPase assays using 32P‐labeled Raytide as a substrate, and that 32P‐labeled p107wee1 is an in vitro substrate for pyp1. To compare the biological activity of pyp1 and pyp2 to that of other known PTPases, we expressed the budding yeast PTP1 and human placental phosphatase 1B (PTP1B) genes in either a cdc25–22 or wee1–50 genetic background and established that, in contrast to pyp1+ and pyp2+, Saccharomyces cerevisiae PTP1 and human PTP1B complement the cdc25 mutant, opposing the wee1+ lmik1+ pathway.
The nucleotide sequence of a 2·1 kb fragment from chromosome VI of Saccharomyces cerevisiae identifies a tRNAGly gene, part of a delta element and a palindromic sequence
Authors: Van Heusden G. Paul H. , De Koning Wim J. , Van Der Aart Quirina J. M. , Van Den Berg Johan A. , Steensma H. Yde 
Published: May 2005

AbstractThe nucleotide sequence was determined of a 2·1 kb DNA fragment located at approximately 35 kb to the right of the centromere of chromosome VI from Saccharomyces cerevisiae. Analysis revealed the presence of a tRNAGLy gene, part of a delta element and a remarkable palindromic sequence. The longest open reading frame found encodes a putative protein of 195 amino acids. Although the fragment was isolated by hybridization to a human diacylglycerol kinase cDNA, no evidence was obtained for the presence of a gene encoding diacylglycerol kinase.
The complete sequence of a 6794 bp segment located on the right arm of chromosome II of Saccharomyces cerevisiae. Finding of a putative dUTPase in a yeast
Authors: Doignon Francois , Biteau Nicolas , Aigle Michel , Crouzet Marc 
Published: May 2005

AbstractThe DNA sequence of a 6794 bp fragment located at about 100 kb from the right telomere of chromosome II from Saccharomyces cerevisiae has been determined. Sequence analysis reveals five open reading frames. One is the ARO4 gene encoding the 3‐deoxy‐D‐arabinoheptulosonate 7‐phosphate synthase. Another presents strong homology with the S5 ribosomal protein from bacteria. The open reading frame YBR1705 shows significant homology with dUTPase, suggesting for the first time the existence of such an enzyme in S. cerevisiae.
The ERG3 gene in Saccharomyces cerevisiae is required for the utilization of respiratory substrates and in heme‐deficient cells
Authors: Smith Steven J. , Parks Leo W. 
Published: May 2005

AbstractERG3 is the structural gene in Saccharomyces cerevisiae for the sterol Δ5 desaturase that introduces the C5=6 unsaturation in ergosterol biosynthesis. The ERG3 gene has been mapped on chromosome XII, 13·7 centimorgans from GAL2 toward SPT8. The essentially of the gene is dependent on the conditions used for the cultivation of the mutants. Insertionally inactivated mutants of ERG3 fail to grow without ‘sparking’ levels of Δ5 sterols in heme‐deficient cells, and are unable to grow on the respiratory substrates glycerol and ethanol.
Transformation in the methylotrophic yeast Pichia methanolica utilizing homologous ADE1 and heterologous Saccharomyces cerevisiae ADE2 and LEU2 genes as genetic markers
Authors: Hiep Tran Tuan , Noskov Vladimir N. , Pavlov Yuri I. 
Published: May 2005

AbstractDevelopment of transformation systems for methylotrophic yeasts is the starting point for research aimed at developing molecular genetics of these genera and will be the key to their further successful use in biotechnology. We transformed Pichia methanolica using selector genes ADE2 and LEU2 from Saccharomyces cerevisiae and ADE1 (homologue of S. cerevisiae ADE2 gene) from P. methanolica which was cloned and sequenced in our laboratory (Hiep et al., 1991). Lithium transformation of P. methanolica strains was inefficient with intact plasmids. Linearization of plasmids at a unique restriction site within the ADE1 gene prior to transformation substantially increased its frequency. Transformation with linear ADE1, ADE2 or LEU2 gene fragments was even more effective. Introduced DNA fragments either circularized in vivo, irrespective of the structures of their ends, giving unstable transformants; or integrated at different sites of the host genome. Using this transformation system, we obtained a disruption of the ADE1 gene on the chromosome by inserting the S. cerevisiae LEU2 gene. The disruption mutation ade1::LEU2 was used to study the mechanism of intragenic recombination in P. methanolica.
Fluorescent staining with bromocresol purple: A rapid method for determining yeast cell dead count developed as an assay of killer toxin activity
Authors: Kurzweilová Helena , Sigler Karel 
Published: May 2005

AbstractA method is described for detecting yeast cells with plasma membrane damage, based on cell staining with bromocresol purple (BCP) which has a convenient fluorescence after entering the cells at pH below 5·2. The method was used to determine the activity of Saccharomyces cerevisiae pore‐forming killer toxin K1 in commonly used lethal units. The BCP test is rapid and as precise as the plating test.
Sequence and function analysis of a 2·73 kb fragment of Saccharomyces cerevisiae chromosome II
Authors: Miosga Thomas , Zimmermann Friedrich K. 
Published: May 2005

AbstractThe nucleotide sequence of a fragment of 2728 base pairs of Saccharomyces cerevisiae chromosome II has been determined. The sequence contains two open reading frames, one of them being incomplete. Deletion mutants of YBR11.21 are viable. YBR11.20 is identical to the recessive omnipotent suppressor SUP45 (SUP1).
A series of yeast shuttle vectors for expression of cDNAs and other DNA sequences
Authors: Brunelli Joseph P. , Pall Martin L. 
Published: May 2005

AbstractExpression/shuttle vectors for the yeast Saccharomyces cerevisiae have usually been large plasmids with only one or a small number of sites that are suitable for cloning and expression. We report here the construction and properties of a series of 12 expression vectors with multiple (four to eight) unique sites in their polylinkers which allow directional cloning and expression of DNA sequences under four different promoters. Eleven of these plasmids replicate at high copy number in Escherichia coli, and all have the yeast TRP1 gene, and the 2 μm origin including REP3 sequence, allowing selection and high copy number replication in yeast. Six of the plasmids are designed for the construction and selection of cDNA libraries from various eukaryotic organisms, allowing directional cloning and expression of cDNAs. All of these six have similar polylinkers containing a unique promoter proximal EcoRI site and a unique promoter distal XhoI site, allowing for directional cloning and expression of ‘ZAP’‐type cDNAs. cDNAs that complement a wide variety of yeast mutants can be selected from libraries constructed in this way. The four alternative promoters, ADH2, PGK, GAL10 and SV40 were compared for their relative activity, both in E. coli and in yeast. All yeast promoters showed substantial activity in E. coli with ADH2 showing the highest activity. ADH2 also was well‐regulated in yeast, showing very high relative activity under derepressing conditions. cDNAs selected by genetic complementation from libraries constructed in these vectors should be easily subclonable into other vectors, allowing expression in different eukaryotic organisms, DNA sequencing or site‐directed mutagenesis.
A series of yeast/Escherichia coli λ expression vectors designed for directional cloning of cDNAs and cre/lox‐mediated plasmid excision
Authors: Brunelli Joseph P. , Pall Martin L. 
Published: May 2005

AbstractA series of Saccharomyces cerevisiae/Escherichia coli λ/plasmid expression vectors have been constructed which allow easy excision of the plasmid sequences from λ. Features of six are described, and two designated λPG15 and λAD5, are characterized in detail. Transcription of cloned sequences is controlled by the alternative promoters, ADH2, PGK, GAL10 and SV40 early, and by the CYC1 transcriptional terminator. Unique EcoRI and XhoI restriction sites in the intervening polylinker make these λ vectors compatible for directional cloning of ‘ZAP’‐synthesized cDNAs. Inserted DNAs have been previously shown to have high levels of the genetic activity in both S. cerevisiae and E. coli, allowing these vectors to be used for genetic complementation in both species. Plasmid recovery from the λ vector is mediated by the activity of the cre‐encoded enzyme upon lox sequences flanking the plasmid and adjoining the λ arms. The plasmids contain the yeast 2 μm origin and E. coli pBR322 origin, the URA3 or TRP1 yeast selectable markers, and ampicillin‐resistance marker in E. coli. The usefulness of the λPG15 and the λAD5 cloning vectors was demonstrated by constructing large Neurospora crassa cDNA libraries. The λPG15–N. crassa library was used to infect purE, purC and trpC mutants of E. coli, and complemented and/or suppressed prototrophic colonies were selected. The flexibility and power of this system for cloning of cDNAs is discussed.
Sequencing and analysis of 51·6 kilobases on the left arm of chromosome XI from Saccharomyces cerevisiae reveals 23 open reading frames including the FAS1 gene
Authors: Wiemann Stefan , Voss Hartmut , Schwager Christian , Rupp Thomas , Stegemann Joseph , Zimmermann Jürgen , Grothues Dietmar , Sensen Christoph , Erfle Holger , Hewitt Neil , Banrevi Andrea , Ansorge Wilhelm 
Published: May 2005

AbstractWe have sequenced two segments containing a total of 51·6 kb of the left arm from chromosome XI of Saccharomyces cerevisiae. The first segment of 38·5 kb contains 18 open reading frames (ORFs) of more than 100 amino acid residues. Five ORFs encode known yeast genes, including the fatty acid synthase gene (FAS1). Three new yeast genes were discovered with homologies to non‐yeast genes and ten new genes without homologies to any known sequences. The second segment of 13 kb contains five ORFs with two known yeast genes and three unknown genes. The sequences from cosmid pUKG041 were obtained entirely with the walking primer strategy resulting in a very low overall sequence redundancy of 2·8 and an average reading length of 443 bases.
Low‐Affinity glucose carrier gene LGT1 of Saccharomyces cerevisiae, a homologue of the Kluyveromyces lactis RAG1 gene
Authors: Prior C. , Fukuhara H. , Blaisonneau J. , Wesolowski‐Louvel M. 
Published: May 2005

AbstractA low‐affinity glucose transporter gene of Saccharomyces cerevisiae was cloned by complementation of the rag1 mutation in a strain of Kluyveromyces lactis defective in low‐affinity glucose transport. Gene sequence and effects of null mutation in S. cerevisiae were described. Data indicated that there are multiple genes for low‐affinity glucose transport.
The sequence of a 17.5 kb DNA fragment on the left arm of yeast chromosome XI identifies the protein kinase gene ELM1, the DNA primase gene PRI2, a new gene encoding a putative histone and seven new open reading frames
Authors: Purnelle Bénédicte , Tettelin Hervé , Van Dyck Luc , Skala Jacek , Goffeau André 
Published: May 2005

AbstractA 17.5 kb DNA fragment of chromosome XI, located between the genetic loci mif2 and mak11 was sequenced and analysed. Ten open reading frames were identified. Two of them are the previously sequenced genes ELM1 and PRI2, two (YKL253 and YKL256) show homologies to proteins from other organisms and one (YKL262) to yeast and mouse histone.
Analysis of the rad3‐101 and rad3‐102 mutations of saccharomyces cerevisiae: Implications for structure/function of rad3 protein
Authors: Montelone Beth A. , Malone Robert E. 
Published: May 2005

AbstractThe mutations rad3‐101 and rad3‐102 (formerly rem1‐1 and rem1‐2) of the essential RAD3 gene of Saccharomyces cerevisiae confer a phenotype of semidominant enhancement of spontaneous mitotic recombination and mutation frequencies, but not extreme sensitivity to ultraviolet (UV) light. These properties differ from the previously published observations of other rad3 mutations, which are very UV‐sensitive but do not alter recombination frequencies significantly. We have located the position of DNA sequence changes from wild‐type RAD3 to the rad3‐101 and rad3‐102 mutations and have demonstrated that these sequence changes are necessary and sufficient to confer the (Rem−) mutant phenotype when transferred into otherwise wild‐type RAD3 plasmids. The Rem− mutations are not located in the same region. It is possible that the two regions of the gene in which these mutations map define portions of the molecule which are in contact when folded in the native configuration. To begin to test this hypothesis, we have constructed two double mutant alleles, one with rad3‐101 and rad3‐102, and one with the UV‐sensitive rad3‐1 mutation and rad3‐102. We find that plasmids carrying these double mutant alleles of RAD3 are no longer able to confer a hyper‐recombinational phenotype and do not complement the UV‐sensitivity of the excision‐defective rad3‐2 allele. We conclude that the double mutant alleles are non‐functional for excision repair, and may be null. We have also constructed new rad3 alleles by oligonucleotide‐directed mutagenesis and have tested their effects on spontaneous mutation and mitotic recombination and on UV repair.