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PubMed2005

DNA typing methods for differentiation of yeasts related to dry-cured meat products.

RFLP analysis of the ITS and 18S rDNA, RAPD-PCR using mini- and microsatellite primers and RFLP analysis of mitochondrial DNA were examined to discriminate yeasts related to dry-cured meat products at species and strain level. Seven species and 35 strains of yeasts usually found in dry-cured meat products were tested. RFLP analysis of the ITS1-5.8S rDNA-ITS2 and 18S rDNA did not allow the separation at species level of all of the species tested. RAPD with a M13 primer was found to be useful for differentiation of Rhodotorula mucilaginosa, Candida zeylanoides, Yarrowia lipolytica, Debaryomyces hansenii and Saccharomyces cerevisiae. However, no differences were observed between Debaryomyces polymorphus and Pichia carsonii. RAPD analysis with microsatellite primers (GACA)(4), (GTG)(5) and (GAC)(5) enabled discrimination at species and strain level. However, the degree of discrimination by means of RAPD-PCR depends highly on the primers used. Thus, the PCR fingerprinting with primer (GACA)(4) enabled a higher level of discrimination than primers (GAC)(5) and (GTG)(5). The RFLP analysis of mtDNA allowed the discrimination at the species and strain level except for R. mucilaginosa, where no polymorphisms were observed in the strains tested. RAPD analysis with primer (GACA)(4) and the restriction analysis of mtDNA used in the present work are useful for the differentiation at species and strain level of yeasts related to dry-cured meat products.

DNA Primers↗
PubMed1980

Effect of acriflavine on the hexokinase isozyme pattern of a yeast, Hansenula jadinii.

Dried cells of a yeast, Hansenula jadinii, that had been cultured aerobically with acriflavine, contained three hexokinase isozymes and metabolized glucose at 0.6 M to produce ATP to phosphorylate nucleotides in the presence of a high concentration of phosphate. Dried cells cultured aerobically without acriflavine contained two hexokinase isozymes and could not metabolize glucose under the same conditions. Two of the isozymes of the yeast cultured with acriflavine were similar to isozymes of the yeast cultured without acriflavine. However, the third isozyme was resistant to a high phosphate concentration and caused regeneration of ATP through glycolysis and phosphorylation of nucleotides.

Acridines↗
PubMed1976

Subcellular localization of diffusible ions in the yeast Saccharomyces cerevisiae: quantitative microprobe analysis of thin freeze-dried sections.

In yeast cells, of which the intracellular potassium had been partly replaced by rubidium or caesium, the intracellular ion distribution was studied by means of energy-dispersive X-ray microanalysis. The cells were rapidly frozen and thin sections were cut at low temperature on a cryo-ultramicrotome without the use of a trough liquid. By this dry cryosectioning procedure, complete retention of the diffusbile ions in the cells was obtained. Unless the sections had been exposed to moisture, no signs of redistribution were apparent. For quantitative determinations a gelatin standard, containing known amounts of the elements of interest, was prepared in the same way as the cells. The concentrations of potassium, rubidium, caesium and chloride in the nucleus, the cytoplasm and the vacuole could be measured. The intracellular distributions of potassium, rubidium and caesium were very similar. The concentrations of these ions in the cytoplasm were about equal to those in the nucleus and twice those in the vacuole. The total concentration in the cytoplasm was 180-190 mmol/kg fresh weight, in the nucleus 190-200 mmol/kg fresh weight and in the vacuole 75-90 mmol/kg fresh weight. The permeability of the yeast cell for chloride is markedly lower than for the cations.

Cell Nucleus↗