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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 298 (1982), S. 391-393 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The S. cerevisiae strain RJB-1 was obtained during a search for nonsense mutations in the strain FM11 (rf. 6). At 37 C, RJB-1 arrested with the budded, mononucleate morphology illustrated in Fig. 1. When crossed with a wild-type strain, RJB-1 yielded a temperature-insensitive diploid, and 42 ...
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; G1 cdc mutants ; tα-factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants in four G1 cdc strains of Saccharomyces cerevisiae were isolated which failed to show division arrest in the presence of α-factor. The cell cycle properties, terminal arrest morphology and mating competence of these mutants at the restrictive temperature were examined. The G1 specific arrest of the cdc 36 and cdc39 mutants is dependent upon the availability of an intact mating factor response system in Mat a cells. Cdc28 and cdc37 mutants exert their cell cycle blocks independently of the mating factor pathway. It is likely that the nature of the primary growth defect in cdc36 and cdc39 mutants is such that the α-factor pathway is activated in the absence of the pheromone at the restrictive temperature and that G1 arrest is a secondary consequence of a non-cycle specific event in such mutants.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 269 (1977), S. 145-147 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] A number of authors5"7 have shown that if one knows the percentage of cells in an asynchronous culture that are past a particular event in the cell cycle, the stage in the cycle that the event occurs can be determined. The execution point of a temperature-sensitive cell cycle mutant can be ...
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 10 (1986), S. 943-945 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; α-factor ; Protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary When Mat a cells are treated with α-factor prior to being protoplasted and fused, the frequency of karyogamy is higher than in unarrested controls.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0983
    Keywords: S. cerevisiae ; Secretion ; Interferon alpha-factor ; Glycosylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Studies with three interferon molecules, IFN-α2, IFN-β 1, and a “hybrid” interferon, IFNX-430 are described which illustrate that both the expression and secretion characteristics of heterologous proteins in yeast cells reflect properties of the proteins themselves. Recombinant DNA techniques have also been used to demonstrate that the efficient processing of mature heterologous proteins from the yeast α factor secretion leader can be affected by sequences on the carboxyl side of the initial cleavage site. Secretion studies with heterologous proteins in S. cerevisiae are aimed at maximising yield, the percentage of extracefular product and correct amino terminus sequence. The results presented here show that all three factors are susceptible to currently unpredictable properties of the foreign sequence. This situation, in turn, means that heterologous proteins can be used as tools in the biochemical dissection of the yeast secretion process.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Protoplast fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The percentage of hybrids formed during protoplast fusion in Saccharomyces cerevisiae is determined by the percentage of protoplasts at the GI/S boundary of the cell cycle.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 11 (1969), S. 785-804 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The physiology of Aspergillus nidulans strain 224 has been studied under conditions of batch- and glucose-limited chemostat-culture and the effect of different steady state growth rates and dissolved oxygen tensions (DOT) examined. Measurements of the specific activities of selected glucose enzymes, the extent of oxygen uptake inhibition by glycolytic inhibitors, and radiorespirometric analyses were made in order to follow the variations in glucose catabolism, which occurred under these conditions. Greatly increased activity of the hexosemonophosphate (HMP) pathway was found during: (i) exponential growth of batch cultures; (ii) at near maximum specific growth rates (μ = 0.072 hr-1) (DOT = 156 mm Hg); and (iii) at low DOT levels (〈30 mm Hg) (μ = 0.050 hr-1) in chemostat cultures. These changes in glucose eatabolism have been discussed in terms of the biosynthetic demands of the fungus under the influence of changing growth pressures. Preliminary studies also have been made of transition state behavior following stepwise alteration of the DOT. A new steady state was established after 4-5 culture doublings during which period an “overshoot” in HMP pathway activity occurred; these kinetics are indicative of a derepression of certain glucose enzymes. Low molecular weight phenols are synthesized during the exponential phase in batch cultures and these are further metabliized to a major secondary metabolite, melanin, at the onset of stationary phase conditions. The kinetics of tyrosinase production in steady state chemostats differs from those that might be predicted for an enzyme associated solely with secondary metabolism. A primary physiological role for this oxidase in Aspergillus nidulans has been postulated.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Yeast ; Copy number ; Thymidine kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 2 μm circle-based chimaeric plasmid containing the yeast LEU2 and the Herpes Simplex Virus type 1 thymidine kinase (HSV-1 TK) genes was constructed. Transformants grown under selective conditions for the LEU2 gene harboured the plasmid at about 15 copies per cell whilst selection for the HSV-1 TK gene led to an increase to about 100 copies per cell. Furthermore, the plasmid copy number could be controlled by the stringency of selection for the TK gene, and the increase in TK gene dosage was reflected in an increase in intracellular thymidine kinase activity. The mitotic stability of the plasmid in “high-copy” and “low-copy” number cells was determined. “High-copy” number cells showed a greater mitotic stability. The relationship of TK expression to plasmid copy number may be useful for the isolation of plasmid copy number mutants in yeast and the control of heterologous gene expression.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1617-4623
    Keywords: Yeast ; HSV TK ; Promoter ; Interferon ; Secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A system is described that facilitates the selection of yeast promoters of unknown origin using a functional heterologous gene assay. A multicopy plasmid was constructed with a unique cloning site 5′ to the Herpes Simplex type 1 thymidine kinase gene (TK). A yeast genomic DNA library was introduced into this site and used to transform yeast (which naturally lacks a TK gene). Only transformants carrying DNA fragments with promoter activity can express TK activity and thereby grow on medium containing thymidine and folate antagonists. On the basis of TK assays of yeast cell homogenates from some 100 TK + transformants, a particularly promising plasmid containing a yeast promoter was selected for further analysis. The position of the transcriptional start of the mRNA of this promoter was identified and the promoter sequence established. A fragment containing the promoter was taken and engineered to produce an expression vector and an expression/secretion vector. We report on the expression and secretion of interferon —X430 (IFN-X 430) (a derivative of IFN beta) from yeast cells containing such plasmids.
    Type of Medium: Electronic Resource
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