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  • 1
    ISSN: 1520-6033
    Source: ACS Legacy Archives
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 613 (1990), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The cellular β-galactosidase activities produced by thelac'Z gene ofEscherichia coli, cloned on YEp, YRp, or YCp-type plasmids in host cells ofSaccharomyces cerevisiae with different ploidies, and which was expressed by a modified jeastHIS5 promoter, showed characteristic differences depending on the plasmid. But for any given plasmid, the isogenic diploid and tetraploid transformants showed slightly lower enzyme activities than their respective haploid transformants. This was due to the similar copy numbers of the plasmids in host cells. Since the cell number per unit volume of the culture decreased with increasing cell ploidy, the enzyme activity per unit volume of the culture decreased significantly. The holding stability of plasmids increased with increasing ploidy of the host cell, especially that of the YRp plasmid. On the YRp plasmid, thelac'Z gene showed higher productivity withTRP1 thanLEU2 as the selection marker for the plasmid.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The cellular β-galactosidase activities produced by thelac'Z gene ofEscherichia coli, cloned on YEp, YRp, or YCp-type plasmids in host cells ofSaccharomyces cerevisiae with different ploidies, and which was expressed by a modified jeastHIS5 promoter, showed characteristic differences depending on the plasmid. But for any given plasmid, the isogenic diploid and tetraploid transformants showed slightly lower enzyme activities than their respective haploid transformants. This was due to the similar copy numbers of the plasmids in host cells. Since the cell number per unit volume of the culture decreased with increasing cell ploidy, the enzyme activity per unit volume of the culture decreased significantly. The holding stability of plasmids increased with increasing ploidy of the host cell, especially that of the YRp plasmid. On the YRp plasmid, thelac'Z gene showed higher productivity withTRP1 thanLEU2 as the selection marker for the plasmid.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 28 (1988), S. 155-159 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A YCp type promoter-probe vector, pVC701, replicable inSaccharomyces cerevisiae andEscherichia coli hosts was constructed. pVC701 has a DNA fragment bearing thePHO5 gene encoding repressible acid phosphatase (rA-Pase; EC 3.1.3.2.) without its promoter region. The clonedPHO5 gene can be expressed by insertion of a DNA fragment having promoter function at theEcoRI site on the 5′-flanking region ofPHO5. rAPase activity caused by thePHO5 expression is easily detected by staining the transformant colonies with diazo-coupling reagent. These were confirmed by insertion of aHIS5 DNA fragment ofS. cerevisiae having promoter function at theEcoRI cloning site in conditions of histidine starvation. Numerous DNA fragments exhibiting promoter function were isolated by employing pVC701. Most of them expressed thePHO5 gene constitutively, while one of them conferred galactose-inducible and glucose-repressible expression.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0983
    Keywords: Key wordsSaccharomyces ; Tup1-Ssn6 repression ; IME1 ; Sin4-Rgr1 repression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ime1 plays a pivotal role in the initiation of meiosis in a/α diploid cells of Saccharomyces cerevisiae. In the absence of glucose and nitrogen, IME1 expression is greater in a/α cells than in either a or α cells and therefore only a/α, but not a/a or α/α, cells are committed to sporulation. It is known that IME1 expression is positively regulated by Mck1, Rim1, Ime4 and the Swi-Snf complex but other factors may also be involved. In addition, Rme1 is assumed to repress IME1 expression. To provide more details of the repression of expression of IME1, we have isolated mutants in which the IME1p-PHO5 fusion gene integrated at the ura3 locus is expressed in α cells under nutritionally rich conditions. We found that mutations occurred in TUP1, SSN6, SIN4 and RGR1, among which TUP1 and SSN6 were identified for the first time as negative regulators of IME1 expression. Deletion of the Rme1-binding site from the IME1 promoter did not result in activation of the expression of IME1 under nutritionally rich conditions, suggesting that Rme1 does not function as a DNA-binding protein with the Tup1-Ssn6 repression complex. We also demonstrated that the 294-bp fragment from nucleotide position –914 to –621 and the 301-bp fragment from nucleotide position –1215 to –915 of the IME1 promoter region contain elements acting as URS and UAS in TUP1 + and tup1 mutant cells, respectively. These findings indicate that IME1 is negatively regulated by the Tup1-Ssn6 repressor complex through two distinct upstream regions in conjunction with unidentified DNA-binding proteins.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0983
    Keywords: Arsenate resistance ; Phosphatase regulation ; Pi-transporter ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract ThePHO84 gene inSaccharomyces cerevisiae encodes a Pi transporter, mutation of which confers constitutive synthesis of repressible acid phosphatase (rAPase), in medium containing repressible amounts of Pi, and an arsenate-resistant phenotype. We selected an arsenate-resistant mutant showing the constitutive synthesis of rAPase on nutrient plates containing 4.5 mM arsenate. This mutant has double mutations designated aspho86 andpho87. The mutant transcribesPHO84 even in the repressible condition but has a severe defect in Pi uptake. The constitutive rAPase+ phenotype of thepho86 pho87 mutant was partially suppressed by an increased dosage of thePHO84 gene. ThePHO87 gene was found to be identical withYCR524, according to the published nucleotide sequence of chromosome III, which encodes a protein of 923 amino-acid residues with a highly charged N-terminal half followed by a C-terminal half consisting of 12 membrane-spanning segments as in Pho84p. These and the other findings suggest that the Pho86p and Pho87p proteins collaborate with Pho84p in Pi uptake.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two mating-type alleles, a and α, are interchangeable with each other due to the specific mutagenic action of the homothallic genes in Saccharomyces. However, a haploid segregant having the α mating-type potency but inconvertible to homothallism by the mutagenic action of the homothallic genes was segregated from a strain of S. diastaticus. The inconvertibility was strictly specific to the α mating-type clone in its pedigree. The genetic analyses of the inconvertible α clones indicated that the inconvertibility was not due to the loss of the specific homothallic genes nor to a specific cytoplasmic inhibitor for the mating-type conversion. The most possible explanation is the presence of an α mating-type allele which is insensitive or resistant to the specific mutagenic action of the homothallic genes.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Wild-type cells of Saccharomyces cerevisiae cultivated in low-Pi medium actively accumulate inorganic phosphate (Pi), while the same cells cultivated in high-Pi medium do not. A recessive constitutive mutant (phoT), for repressible acid phosphatase (EC 3.1.3.2) synthesis, is described. It shows severely reduced potency of Pi uptake, while the recessive constitutive mutants, phoR and phoU, in the same system show wild-type potency as.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary When the pH of growth medium containing a limited amount of inorganic phosphate is kept below 3.0, cells of Saccharomyces cerevisiae produce repressible alkaline phosphatase but no repressible acid phosphatase. The same cells produce acid phosphatase immediately on shifting the medium pH to 4.0 or above. Like intact cells, spheroplasts prepared from cells grown at pH 3.0 or 4.5 in medium with a limited amount of inorganic phosphate in suspension begin production of acid phosphatase immediately after pH shift from below 3.0 to 4.0, whereas spheroplasts from cells grown in inorganic phosphate-rich medium showed a prolonged lag period (3 h). The enzyme formation on the pH shift was sensitive to cycloheximide. No significant differences could be detected in cellular growth or in incorporation of 3H-L-lysine or 14C-adenine between cells cultivated at pH 3.0 and 4.5. These results along with the fact that the expression of structural genes of repressible acid and alkaline phosphatases is controlled by a common genetic regulatory system, at least in part, indicate that the genetic regulatory system operates to express the structural genes even at low pH, though the expression of repressible acid phosphatase is interrupted. Coupled experiments of temperature and pH shifts with the temperature-sensitive mutants of the regulatory genes suggest that the acidic pH affects the function of the cytoplasmic products of those genes in the expression of the structural gene. Based on these observations, a revised model involving the simultaneous functioning of the regulatory factors was suggested for the genetic regulation of repressible acid phosphatase senthesis.
    Type of Medium: Electronic Resource
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