• Title/Summary/Keyword: yeast mutants

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Characterization of Yellow Mutants Isolated from the Red Yeast Phaffia rhodozyma (Xanthophyllomyces dendrorhous)

  • An, Gil-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.110-115
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    • 1996
  • Yellow mutants of the astaxanthin producing red yeast Phaffia rhodozyma were obtained by nitrosoguanidine mutagenesis. The carotenoid composition of the yelow mutants, Yan-1 and Ny-1, was mainly $\beta$ -carotene (> 95$%$) and torulene (< 5$$). Therefore, the yellow mutants are carotene oxygenation deficient mutants (CODMs). CODMs produced decreased quantities of carotenoids compared to their red parents and this indicated that carotene may regulate its synthesis. CODMs, Yan-1 and Ny-4, on plates containing 50 $\mu$ M antimycin, showed highly pigmented vertical papillae. Antimycin-induced mutants purified from the papillae showed increases in carotenoid content (up to 1 mg $\beta$-carotene/g yeast). CODMs, Yan-1 and Ay-1, were more sensitive to antimycin than red strains, Ant-1 and 67-385. This was probably due to lower antioxidant activity of $\beta$-carotene than that of astaxanthin. Light increased torulene and light+antimycin further increased the torulene. Yan-1 and Ny-4 could grow with succinate, though their red parents, Ant-1 and Anf-1p, could not. However, antimycin induced mutation of Yan-1 or Ny-4 destroyed the ability to grow with succinate.

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Isolation and Characterization of Salt Tolerant Mutations in Budding Yeast Saccharomyces cerevisiae

  • Kim, Yung-Jin;Seo, Soo-Boon;Park, Shi-Young
    • Journal of Life Science
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    • v.9 no.1
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    • pp.22-25
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    • 1999
  • In order to study the mechanism for the adaptation to salt stress, we mutagenized budding yeast Saccharomyces cerevisiae with Ethylmethane sulfonate, and isolated salt-tolerant mutants. Among the salt-tolerant mutants, two strains exhibit additional temperature sensitive phenotype. Here, we report that these two salt-tolerant mutants are specific to {TEX}$Na^{+}${/TEX} rather than general osmotic stress. These mutant strains may contain mutations in the genes involved in {TEX}$Na^{+}${/TEX} home-ostasis.

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Selection of Enhanced Iron Uptake Yeast Mutants by EMS Mutagenesis. (EMS에 의하여 철 함유 능력이 증진된 효모 돌연변이주의 선별)

  • 양승남;송형석;이중림;김해영
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.435-438
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    • 2003
  • Iron required by all organisms is related with diverse biological processes. Most eukaryotes need extra iron to maintain their nutrition balance. However, extra iron supplement gives many problem to solubility in the cells. To increase the bio-availability of iron in cells, yeast was applied to carry the iron with solubility. Selection of yeast mutants with enhanced iron uptake were performed by mutagenesis using the alkylation agent EMS. Eleven mutant strains with enhanced iron uptake were selected by the measurement of iron content with atomic absorption spectrometer. The iron content in mutants was 1.5- to 2.5-fold more than that in wild-type. These mutants could be served as iron-fortified nutrients for food and feed.

A pheromone mutant of Schizosaccharomyces pombe displays nucleolar fragmentation

  • Jun, Jai-Hyun;Kim, Dae-Myung
    • BMB Reports
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    • v.41 no.3
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    • pp.248-253
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    • 2008
  • Stresses and nutritional starvation are two main external signals for the induction of sex pheromones in the fission yeast Schizosaccharomyces pombe. In an attempt to identify the components involved in transduction of starvation signals, we screened 135 temperature-sensitive (ts) mutants and isolated 6 mutants that induced the pheromone even in the presence of a nitrogen source. These mutants exhibited two distict induction phenotypes: pheromone induction at restrictive but not at permissive temperatures; and pheromone induction at both permissive and restrictive temperatures. The times required for the maximum pheromone induction at the restrictive temperature differed slightly in each mutant. In addition to the pheromone induction phenotype, the ts243 and ts304 mutants exhibited cell-division-cycle defects. The ts304 mutant cells showed an abnormal cytoplasmic DAPI staining pattern. The nucleolus of this mutant seemed to be fragmented, a phenomenon which is typically observed in aged yeast cells. The result of our genetic analysis indicated that the pheromone induction mutants belonged to 6 separate complementation groups. We designated these mutants pws1 to pws6.

Isolation and Characterization of Salt Street Signaling Components from Yeast Saccharomyces cerevisiae

  • Yun, Dae-Jin;Lee, Jiyoung;Shin, Dongjin;Lee, Boyoung
    • Journal of Life Science
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    • v.11 no.1
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    • pp.30-33
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    • 2001
  • To identify novel components involved in the salt stress signaling pathway of yeast cells, we used mTn3-mediated transposon tagging library and screened mutants displaying enhanced tolerance to NaCl. Southern blot analysis indicated that more than 80% of the sre (salt resistant) mutants possessed only one insertion of the tagged transposon, suggesting that the NaCl resistant phenotype was mediated by a single gene in the majority of the mutants. To define the role of SRE genes in the salt stress signaling pathway, we introduced NaCl stress-inducible ENA1::LacZ construct into the sre mutants and examined the expression of ${\beta}$-galactosidase activity. Interestingly, we could detect high level of ${\beta}$-galactosidase activity without any NaCl treatment in the sre-3, 4, 6 and 7 mutants. These results indicate that SRE-3, 4, and 7 gene are components of salt stress signaling pathway of yeast cells.

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Immune-Enhancing Alkali-Soluble Glucans Produced by Wild-Type and Mutant Saccharomyces cerevisiae

  • Ha Chang-Hoon;Lim Ki-Hong;Jang Se-Hwan;Yun Cheol-Won;Paik Hyun-Dong;Kim Seung-Wook;Kang Chang-Won;Chang Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.576-583
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    • 2006
  • The alkali-soluble glucan of the yeast cell wall contains $\beta-(1,3)-$ and (1,6)-D-linkages and is known to systemically enhance the immune system. In the previous study [6], in order to isolate cell wall mutants, a wild-type strain was mutagenized by exposure to ultraviolet light, and the mutants were then selected via treatment with laminarinase $(endo-\beta-(1,3)-D-glucanase)$. The mass of alkali- and water-soluble glucans produced by the mutant was measured to be 33.8 mg/g of the dry mass of the yeast cell. Our results showed that the mutants generated the amount of alkali-soluble glucan 10-fold higher than that generated by the wild-type. Structural analysis showed that the alkali-soluble glucan from the mutants was associated with a higher degree of $\beta-(1,6)-D-linkage$ than was observed in conjunction with the wild-type. Yeast cell wall $\beta-glucan$ was shown to interact with macrophages via receptors, thereby inducing the release of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide. Alkali-soluble $\beta-glucans$, both from water-soluble and water-insoluble glucan, exhibited a higher degree of macrophage activity with regard to both the secretion of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide and direct phagocytosis, than did the positive control ($1{\mu}g$ of lipopolysaccharide).

Selection of Nitrate-nonutilizing Mutants of Hypoxylon atropunctatum, A Fungal Pathogen on Oak Species

  • Chun, Se-Chul;Fenn, P.
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.247-251
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    • 2000
  • Latent infections of healthy-appearing oaks of Hypoxylon atropunctatum complicates field studies by interfering with inoculation experiments to follow pathogenesis, fungal development and reproduction of this canker rot fungus. Mutants with unique and easily scorable phenotypes would be useful for inoculation studies. There is a broad range in the capacity of wild-type isolates to utilize nitrate as a sole nitrogen sources. Several types of nitrate-nonutilization mutants (nit1, Nit3, NitM) were selected from nitrate-utilizing wild-type isolates. Also, a few mutants of Hypoxylon atropunctatum were selected that could only grow poorly on basal medium supplemented with various nitrogen sources and even on yeast extract agar. These unknown mutants need to be characterized further. Nit mutants of Hypoxylon atropunctatum were readily selected, grew well and were recovered after inoculation into oak stems. These results suggest that nit mutants could be useful for inoculation studies in trees that contain latent infections.

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A yeast Chromosomal Gene that Induces Defective Interfering Particles of L-A dsRNA Virus in $ski^-$ Host Cells ($ski^-$ 기주 세포에서 L-A dsRNA 바이러스의 defective interfering particle을 유도하는 효모 유전자)

  • 이현숙
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.75-79
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    • 1991
  • The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.

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