Selection of auxotrophic mutants inSaccharomyces cerevisiae by a snail enzyme digestion method

Piedra, D; Herrera, Luís

doi:10.1007/bf02876959

Cited by 11 publications

(4 citation statements)

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“…Several lines of evidence suggest that stationary-phase cells are in a physiological state qualitatively different from that of cycling cells. Quiescent cells display resistance to killing by heat (29,33) and by cell wall-degrading enzymes (5,25). They differ also from cycling cells by their RNase activity (22,32), polyadenylate content (30), and the structure of their folded genomes (26,27).…”

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Protein synthesis during transition and stationary phases under glucose limitation in Saccharomyces cerevisiae

Boucherie¹

1985

J Bacteriol

102

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Metabolic changes have been investigated during continuous growth of yeast cells inoculated in glucose-containing medium until the ceUls entered the stationary phase in response to glucose exhaustion. Well in advance of glucose exhaustion, a transition phase was observed, characterized by a decrease in the growth rate and a progressive reduction of protein and RNA accumulation, Two-dimensional gel analysis of the proteins synthesized during this stage showed that the pattern of proteins remained similar to that of log-phase ceUls. When the cells entered the stationary phase, protein accumulation was 10% of that in log-phase cells, and incorporation of labeled RNA precursor was undetectable. Analysis of protein synthesis gave evidence that the synthesis of 95% of the proteins present in log-phase ceUls was arrested in stationary-phase cells. Among the 20 proteins whose synthesis continues throughout the stationary phase were identified actin, aldehyde dehydrogenase, enolase, hexokinase, glyceraldehyde-3-phosphate dehydrogenase, and five heat shock proteins. In addition, the synthesis of six new proteins was observed. The occurrence of these new proteins in stationary-phase ceUs is presumed to result from the release of carbon catabolite repression due to glucose exhaustion.

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confidence: 99%

Protein synthesis during transition and stationary phases under glucose limitation in Saccharomyces cerevisiae

Boucherie¹

1985

J Bacteriol

102

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“…Exponential phase cells are sensitive to cell walkdegrading enzymes, whereas stationary phase cells are quite resistant (Piedra & Herrera, 1976). Cells cultured at different mass doubling times in the chemostat were treated with cell wall-degrading enzymes and the percentage of cells in the population that survived the treatment was noted ( Table 1).…”

Section: Characteristics Of Phase-light Cells From Chemostat Culturementioning

confidence: 99%

Differentiation of Saccharomyces cerevisiae at Slow Growth Rates in Glucose-limited Chemostat Culture

1982

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~~ ~Saccharomyces cerevisiae was grown in a glucose-limited chemostat at dilution rates equivalent to mass doubling times of 2 to 16 h. Phase contrast microscopy revealed that cultures with mass doubling times greater than 4.5 h contained two sub-populations of cells, phase-dark and phaselight. The latter were shown to be predominantly unbudded daughter cells, which were not proliferating, and had properties similar to those of stationary phase cells in batch culture. Phase-dark cells were equivalent to cells growing exponentially in batch culture, The results suggest that the formation of phase-light cells represents cell cycle specific differentiation into stationary phase. The consequences of this phenomenon for the interpretation of results obtained with S. cerevisiae in the chemostat are discussed.

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“…Many other methods are based on selective killing of prototrophs (see Fincham et al, 1979). When techniques for the isolation of protoplasts became available, cell wall degrading enzymes were proposed as tools for the isolation of auxotrophic mutants (Delgado et al 1979;Ferenczy et al 1975;Piedra and Herrera 1976;Sipiczky and Ferenczy 1978). However, enrichment of auxotrophs by such enzymes has mainly been applied to yeasts and not to filamentous fungi.…”

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Mitotic mapping in linkage group V of Aspergillus niger based on selection of auxotrophic recombinants by Novozym enrichment

Debets

Swart²,

Bos³

1989

Can. J. Microbiol.

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This paper describes a procedure which allows the quantitative selection of auxotrophs of the fungus Aspergillus niger by enzymatic killing of immobilized germinating prototrophic conidiospores. We have applied this procedure to linkage analysis on the basis of mitotic cross-over in this fungus. Starting with a heterozygous diploid strain, we could select auxotrophic homozygous diploid recombinants quantitatively. We estimated the frequency of crossing-over after correction for clonal distribution of recombinants, and localized four auxotrophic markers as well as the centromere on chromosome V of this fungus. The Novozym enrichment procedure proved to be useful in genetic analysis and for the construction of recombinant genotypes in the case of closely linked auxotrophic markers. The determination of gene order and the estimation of distances on the basis of benomyl-induced recombinant haploid segregants may lead to incorrect conclusions. Genetic analysis on the basis of homozygous recombinants, however, can provide reliable estimates of map distances.

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Selection of auxotrophic mutants inSaccharomyces cerevisiae by a snail enzyme digestion method

Cited by 11 publications

References 11 publications

Protein synthesis during transition and stationary phases under glucose limitation in Saccharomyces cerevisiae

Protein synthesis during transition and stationary phases under glucose limitation in Saccharomyces cerevisiae

Differentiation of Saccharomyces cerevisiae at Slow Growth Rates in Glucose-limited Chemostat Culture

Mitotic mapping in linkage group V of Aspergillus niger based on selection of auxotrophic recombinants by Novozym enrichment

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