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  • 1
    Electronic Resource
    Electronic Resource
    Cloning and disruption of the LEU2 gene of Kluyveromyces marxianus CBS 6556 (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 7 (1991), S. 963-970 
    Details
    ISSN: 0749-503X
    Keywords: Kluyeromyces marxianus ; LEU2 gene ; β-isopropylmalate dehydrogenase ; leu2 mutant ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The LEU2 gene, coding for β-isopropylmalate dehydrogenase, of the yeast Kluyveromyces marxianus was isolated and sequenced. An open reading frame, coding for a protein with a molecular weight of 38kDa was found. Comparison of the deduced amino acid sequence of the LEU2 gene with the corresponding enzymes of three other yeasts and two thermophilic bacteria, revealed extensive sequence similarities. The cloned gene could complement a leuB mutation of Escherichia coli and a leu2 mutation of Saccharomyces cerevisiae. Using orthogonal field alternation gel electrophoresis, the genomic copy of the gene was found to be located at chromosome VI or VII. Analysis of the 5′-untranslated region indicated the presence of a putative binding site for the LEU3 protein, which is involved in the leucine-specific regulation of transcription. We show that the cloned gene can be used for the construction of a non-reverting K. marxianus leu2 mutant.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320070908
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  • 2
    Electronic Resource
    Electronic Resource
    Control of cAMP-induced gene expression by divergent signal transduction pathways (1991)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 12 (1991), S. 25-34 
    Details
    ISSN: 0192-253X
    Keywords: Dictyostelium ; stimulation kinetics ; aggregation-related genes ; prestalk-related genes ; prespore genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A compilation of literature data and recent experiments led to the following conclusions regarding cyclic adenosine 3′:5′ monophosphate (cAMP) regulation of gene expression. Several classes of cAMP-induced gene expression can be discriminated by sensitivity to stimulation kinetics. The aggregation-related genes respond only to nanomolar cAMP pulses. The prestalk-related genes respond both to nano-molar pulses and persistent micromolar stimulation. The prespore specific genes respond only to persistent micromolar stimulation.The induction of the aggregation- and prestalk-related genes by nanomolar cAMP pulses may share a common transduction pathway, which does not involve cAMP, while involvement of the inositol 1,4,5-trisphosphate (IP3)/Ca2+ pathway is unlikely. Induction of the expression of prespore and prestalk-related genes by micromolar cAMP stimuli utilizes divergent signal processing mechanisms. cAMP-induced prespore gene expression does not involve cAMP and probably also not cyclic guanosine 3′.5′ monophosphate (cGMP) as intracellular intermediate. Involvement of cAMP-induced phospholipase C (PLC) activation in this pathway is suggested by the observation that IP3 and 1,2-diacylglycerol (DAG) can induce prespore gene expression, albeit in a somewhat indirect manner and by the observation that Li+ and Ca2+ antagonists inhibit prespore gene expression. Cyclic AMP induction of prestalk-related gene expression is inhibited by IP3 and DAG and promoted by Li+, and is relatively insensitive to Ca2+ antagonists, which indicates that PLC activation does not mediate prestalk-related gene expression. Neither prespore nor prestalk-related gene expression utilizes the sustained cAMP-induced pHi increase as intracellular intermediate.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020120107
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  • 3
    Electronic Resource
    Electronic Resource
    Hemochromatosis and pyruvate kinase deficiency (1991)
    Springer
    Annals of hematology 62 (1991), S. 188-189 
    Details
    ISSN: 1432-0584
    Keywords: Hemochromatosis ; Pyruvate kinase deficiency ; Hereditary anemia ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Hemochromatosis has been reported in several patients with chronic hemolytic anemia due to pyruvate kinase deficiency. We describe here a further patient with such an association and review the literature on the subject. We hypothesize that iron overload may occur in patients with pyruvate kinase deficiency who are also carriers of the hereditary hemochromatosis gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01703147
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  • 4
    Electronic Resource
    Electronic Resource
    Association between a restriction fragment length polymorphism at the liver/islet cell (GluT 2) glucose transporter and familial Type 2 (non-insulin-dependent) diabetes mellitus (1991)
    Springer
    Diabetologia 34 (1991), S. 734-736 
    Details
    ISSN: 1432-0428
    Keywords: Genetics ; diabetes mellitus ; restriction fragment length polymorphism ; glucose-transport ; familial
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Patients with Type 2 (non-insulin-dependent) diabetes mellitus and a strong family history of the disease may represent a sub-group where genetic factors play a pree-minent role in transmission of the disease. A defect in the liver/islet cell glucose transporter (GluT 2) could explain many of the pathophysiological features of the disease. In order to test the hypothesis that genetic variation at the GluT 2 locus contributes genetic susceptibility to Type 2 diabetes, 60 unrelated Caucasian diabetic patients with at least one affected sibling were genotyped for a Taq 1 restriction fragment length polymorphism marker. Hybridisation with a cDNA GluT 2 probe identified two alleles of sizes 13 kilobase (T1) and 19 kilobase (T2). The allele frequencies in the diabetic group with a family history were significantly different from those in a racially-matched control population of 122 subjects with no personal or family history of the disease (diabetic patients T1=0.96, T2=0.04, control subjects T1=0.89, T2=0.11, p〈 0.03). However, when the study was repeated with 54 diabetic patients with indeterminate family history, statistical significance was not reached although the allele frequencies showed a similar trend. The findings of this study support the hypothesis that a genetic variant of the liver/islet cell glucose transporter may contribute to familial susceptibility in Type 2 diabetes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00401519
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  • 5
    Electronic Resource
    Electronic Resource
    Molecular heterogeneity and genetics of Vicia faba seed storage proteins (1991)
    Springer
    Theoretical and applied genetics 81 (1991), S. 50-58 
    Details
    ISSN: 1432-2242
    Keywords: Vicia faba ; Legumin ; Vicilin ; Structure ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Legumin and vicilin were purified from seeds of Vicia faba L. var. Scuro, characterized in different electrophoretic systems, and used to produce polyclonal antibodies in rabbits. Two-dimensional electrophoretic studies showed a wide range of heterogeneity in the subunits of both legumin and vicilin. Legumin was found to be composed of 29 disulphide-linked subunit pairs with different molecular weight and/or isoelectric point. Western blot analysis of legumin of several mutants revealed molecular polymorphism based on a corresponding gene family. Three different α-major legumin patterns were found, and inheritance studies showed that the 34.3-kD legumin polypeptide is the product of one locus, Lg-1α, which is the first legumin genetic locus described in Vicia faba. Vicilin was found to be composed of as many as 59 subunits distributed in a molecular weight range of 65.7 to 42.8 kD (major polypeptides) and 37.2 to 15.2 kD (minor polypeptides), with different isoelectric points. A model is proposed that explains the possible formation of the minor subunits and the major subunits of 48.2 and 46 kD molecular weight (MW) from proteolytic cleavages and/or glycosilation of precursor polypeptides. Ten different vicilin electrophoretic patterns were observed among the analyzed accessions, which showed large molecular polymorphism that proved to be under genetic control.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00226111
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  • 6
    Electronic Resource
    Electronic Resource
    α-Amylase structural genes in rye (1991)
    Springer
    Theoretical and applied genetics 82 (1991), S. 771-776 
    Details
    ISSN: 1432-2242
    Keywords: Secale cereale ; RFLP ; α-Amylase ; Genetics ; Isozymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Rye α-Amy1, α-Amy2, and α-Amy3 genes were studied in the cross between inbred lines using wheat α-amylase cDNA probes. The α-Amy1 and α-Amy2 probes uncovered considerable restriction fragment length polymorphism, whereas the α-Amy3 region was much more conserved. The numbers of restriction fragments found and the F2 segregation data suggest that there are three α-Amy1 genes, two or three α-Amy2 genes, and three α-Amy3 genes in rye. These conclusions were supported by a simultaneous study of α-amylase isozyme polymorphism. The F2 data showed the three individual α-Amy1 genes to span a distance of 3cM at the locus on chromosome 6RL. The genes were mapped relative to other RFLP markers on 6RL. On chromosome 7RL two α-Amy2 genes were shown to be separated by 5 cM. Linkage data within α-Amy3 on 5RL were not obtained since RFLP could be detected at only one of the genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00227324
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  • 7
    Electronic Resource
    Electronic Resource
    Influence of the codon following the initiation codon on the expression of the lacZ gene in Saccharomyces cerevisiae (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 7 (1991), S. 157-165 
    Details
    ISSN: 0749-503X
    Keywords: Translation initiation ; codon usage: mRNA structure ; yeast ; lacZ fusion protein ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A set of 32 different codons were introduced in a lacZ experssion vector (pPTK400) immediately 3′ from the AUG initiation codon. Expression of the lacZ gene was determined in Saccharomyces cerevisiae by measuring the amount of β-galactosidase fusion protein using immuno-gel electrophoresis. A 5·3-fold difference in expression was found among the various constructs. It was found that there was no preference for a certain nucleotide in any position of the second codon and there was no distinct correlation between the level of tRNA corresponding to any particular second codon and expression. No correlation could be found between the local secondary structure and expression. When the overall codon usage in yeast and the codon usage in the second position of the mRNA is compared, there is no obvious significant difference in preference. This indicates that in yeast, in contrast to Escherichia coli, the codon choice at the beginning of the mRNA does not deviate from the one further downstream and is determined by the requirements for optimal translation elongation. Important determinatnts of the optimal context for an initiation codon in yeast therfore must be located mainly 5′ from this codon.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320070209
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  • 8
    Electronic Resource
    Electronic Resource
    Quantitation of readthrough of termination codons in yeast using a novel gene fusion assay (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 7 (1991), S. 173-183 
    Details
    ISSN: 0749-503X
    Keywords: Translation ; Saccharomyces cerevisiae ; lacZ fusion ; termination ; nonsense suppression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A simple quantitative in vivo assay has been developed for measuring the efficiency of translation of one or other of the three termination codons, UAA, UAG and UGA in Saccharomyces cerevisiae. The assay employs a 3-phosphoglycerate kinase-β-galactosidase gene fusion, carried on a multicopy plasmid, in which the otherwise retained reading frame is distupted by one or other of the three termination codons. Termination readthrough is thus quantitated by measuring β-galactosidase in transformed strains. Using these plasmids to quantitate the endogenous levels of termination readthrough we show that readthrough of all three codons can be detected in a non-suppressor (sup+) strain of S. cerevisiae. The efficiency of this endogenous readthrough is much higher in a [psi+] strain than in a [psi-] strain with the UGA codon being the leakiest in the nucleotide context used. The utility of the assay plasmids for studying genetic modifiers of nonsense suppressors is also shown by their use to demonstrate that the cytoplasmic genetic determinant [pse+] broadens the decoding properties of a serine-inserting UAA suppressor tRNA (SUQ5) to allow it to translate the other two termination codons in the order of efficiency UAA 〉 UAG 〉 UGA.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320070211
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  • 9
    Electronic Resource
    Electronic Resource
    Expression of the α-galactosidase from Cyamopsis tetragonoloba (guar) by Hansenula polymorpha (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 7 (1991), S. 463-473 
    Details
    ISSN: 0749-503X
    Keywords: Secretion ; methylotrophic yeast ; glycosylation ; methanol oxidase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The methylotrophic yeast Hansenula polymorpha, a host organism for the production of heterologous proteins, has been applied to produce the α-galactosidase from the plant Cyamopsis tetragonoloba (guar). The yeast/Escherichia coli shuttle expression vector used is based on the origin of replication of the endogenous 2 μm plasmid of Saccharomyces cerevisiae and the LEU2 gene of S. cerevisiae for selection in H. polymorpha. In the expression vector, the α-galactosidase is controlled by the methanol-regulated promoter from the methanol oxidase gene, MOX, of H. polymorpha. The signal sequence of SUC2 (invertase) from the yeast S. cerevisiae, was used to ensure secretion of the α-galactosidase enzyme. After transformation and stabilization, the expression vector was stably integrated in the genome. The active α-galactosidase enzyme was efficiently secreted (〉85%) and after methanol induction, the expression level was 42 mg/l. Amino-terminal sequencing of the purified α-galactosidase enzyme synthesized by H. polymorpha showed that the S. cerevisiae invertase signal sequence was correctly processed by H. polymorpha. The secreted α-galactosidase was glycosylated and had a sugar content of 9·5%. The specific activity of the α-galactosidase produced by H. polymorpha was 38 U mg-1 compared to 100 U mg-1 for guar α-galactosidase. Deglycosylation of the H. polymorpha α-galactosidase restored the specific activity completely.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320070505
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  • 10
    Electronic Resource
    Electronic Resource
    Cyclic variations in the permeability of the cell wall of Saccharomyces cerevisiae (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 7 (1991), S. 589-598 
    Details
    ISSN: 0749-503X
    Keywords: Cell wall porosity ; cell cycle ; centrifugal elutriation ; synchronous growth ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: To study cell-cycle-related variations in wall permeability of Saccharomyces cerevisiae, two approaches were used. First, an asynchronous culture was fractionated by centrifugal elutriation into subpopulations containing cell of increasing size. The subpopulations represented different stages of the cell cycle as judged by light microscopy. Cell wall porosity increased when these subpopulations became enriched with budded cells. Secondly, synchronous cultures were obtained by releasing MATa cells from alpha-factor induced G1-arrest. These cultures grew synchronously for at least two generations. The cell wall porosity incresed sharply in these cultures, shortly before buds became visible and was maximal during the initial stages of bud growth. It decreased in cells which had completed nuclear migration and before abscission of the bud had occurred. The porosity reached its lowest value during abscission and in unbudded cells.We examined the incorporation of mannoproteins into the wall during the cell cycle. SDS-extractable mannoproteins were incorporated continuously. However, the incorporation of glucanase-extractable mannoproteins, which are known to affect cell wall porosity, showed cyclic oscillations and reached its maximum after nuclear migration. This coincided with a rapid decrease in cell wall porosity, indicating that glucanase-extractable mannoproteins might contribute to this decrease.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320070606
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