• Title/Summary/Keyword: yeast mutants

Search Result 144, Processing Time 0.028 seconds

Isolation of Conditional Lethal Temperature-sensitive Mutants of Bacillus sphaericus (Bacillus sphaericus의 치사감온성 돌연변이체의 분리)

  • Kim, Young Han;Lee, Hyung Hoan
    • Microbiology and Biotechnology Letters
    • /
    • v.13 no.1
    • /
    • pp.41-49
    • /
    • 1985
  • Bacillus sphaericus was mutagenized with UV light irradiation and dimethyl sulfate. Thirty-five conditional lethal temperature-sensitive(ts) mutants were isolated at the nonpermissive temperature of $42^{\circ}C$ and classified into three groups by their growth characteristics on the nutrient broth, peptone glucose yeast extract agar and mineral salts agar. First was the lethal ts group, 24 mutants, which did not grow at the nonpermissive temperature, the second, 9 mutants, was the less growth is group whose growth was restricted to one-half, and the third, 2 mutants, was the cold lethal ts group whose growth was restricted at the permissive temperature($25^{\circ}C$and $30^{\circ}C$)

  • PDF

Methionine Analogue Probes Functionally Important Residues in Active Site of Methionyl-tRNA Synthetase

  • Jo, Yeong-Joon;Lee, Sang-Won;Jo, Myung-Kyun;Lee, Jee-Woo;Kang, Mee-Kyoung;Yoon, Jeong-Hyeok;Kim, Sung-Hoon
    • BMB Reports
    • /
    • v.32 no.6
    • /
    • pp.547-553
    • /
    • 1999
  • Aminoacyl-tRNA synthetases are essential enzymes catalyzing the attachment of specific amino acids to cognate tRNAs. In the present work, the substrate analogue L-methionine hydroxamate was used to identify functional residues located in the active site of the E. coli methionyl-tRNA synthetase (MetRS). This compound inhibited bacteria, yeast, and human MetRS activities to a similar degree, suggesting a conserved active site structure and mechanism between MetRSs of different phylogenetic domains. Mutants of the E. coli MetRS resistant to methionine hydroxamate were also isolated. These mutants contained a substitution either at T10, Y15, or Y94. These residues are highly conserved among the different MetRSs and the mutants showed decreased aminoacylation activity, suggesting their functional and structural significances. The putative roles of these residues are discussed on a structural basis.

  • PDF

Effect of Medium Composition on the Bacterial Cellulose Production by Gluconacetobacter hansenii PJK (배지조성이 Gluconacetobacter hansenii PJK의 Bacterial Cellulose 생산에 미치는 영향)

  • 정재용;박연희;박중곤
    • KSBB Journal
    • /
    • v.18 no.2
    • /
    • pp.94-99
    • /
    • 2003
  • The effect of medium composition on the production of bacterial cellulose (BC) by Gluconacetobacter hansenii PJK was investigated. The addition of yeast extract and peptone in the medium increased the production yield (Y/sub p/s/) of BC. The amount of BC produced by G. hansenii PJK was constant if the initial pH of the medium was in the range 4.5 to 6.0. Strains from the supernatant of the culture medium produced more BC than those from inside the BC. BC production was dependent on glucose metabolism, and the addition of fructose or lactate as a carbon source converted cells to Cel/sup -/ mutants. Cel/sup -/ mutants produced by the addition of fructose or lactate to the medium caused 73% or 30% decreases in BC production, respectively. The addition of succinate, which is one of the constituents of the TCA cycle, did not affect the production of BC.

Mitochondrial Porin Isoform AtVDAC1 Regulates the Competence of Arabidopsis thaliana to Agrobacterium-Mediated Genetic Transformation

  • Kwon, Tackmin
    • Molecules and Cells
    • /
    • v.39 no.9
    • /
    • pp.705-713
    • /
    • 2016
  • The efficiency of Agrobacterium-mediated transformation in plants depends on the virulence of Agrobacterium strains, the plant tissue culture conditions, and the susceptibility of host plants. Understanding the molecular interactions between Agrobacterium and host plant cells is crucial when manipulating the susceptibility of recalcitrant crop plants and protecting orchard trees from crown gall disease. It was discovered that Arabidopsis voltage-dependent anion channel 1 (atvdac1) mutant has drastic effects on Agrobacterium-mediated tumorigenesis and growth developmental phenotypes, and that these effects are dependent on a Ws-0 genetic background. Genetic complementation of Arabidopsis vdac1 mutants and yeast porin1-deficient strain with members of the AtVDAC gene family revealed that AtVDAC1 is required for Agrobacterium-mediated transformation, and there is weak functional redundancy between AtVDAC1 and AtVDAC3, which is independent of porin activity. Furthermore, atvdac1 mutants were deficient in transient and stable transformation by Agrobacterium, suggesting that AtVDAC1 is involved in the early stages of Agrobacterium infection prior to transferred-DNA (T-DNA) integration. Transgenic plants overexpressing AtVDAC1 not only complemented the phenotypes of the atvdac1 mutant, but also showed high efficiency of transient T-DNA gene expression; however, the efficiency of stable transformation was not affected. Moreover, the effect of phytohormone treatment on competence to Agrobacterium was compromised in atvdac1 mutants. These data indicate that AtVDAC1 regulates the competence of Arabidopsis to Agrobacterium infection.

Screening and Characterization of Red Yeast Xanthophyllomyces dendrorhous Mutants

  • Kim, Jeong-Hwan;Kim, Chan-Wha;Chang, Hyo-Ihl
    • Journal of Microbiology and Biotechnology
    • /
    • v.14 no.3
    • /
    • pp.570-575
    • /
    • 2004
  • Three different strains of carotenoid accumulating XantlwphyUomyces dendrorhous mutants, JH1, JH2, and JH3, were isolated by NTG (N-methyl-N'-nitro-N-nitrosoguanidine) mutagenesis, which might potentially be useful for animal feed as well as for studies on the regulation and biosynthesis of astaxanthin. Mutants were selected based on the capability of growth and carotenoid production on the YM agar plate containing chemical inhibitor, $\beta$-ionone. Astaxanthin-overproducing mutant JH1 produced 4.032 mg astaxanthinlg dry cell weight, and this value was about 15-folds higher than that of wild-type. $\beta$-Carotene-overproducing mutant JH2 produced 0.273 mg $\beta$-carotene/g dry cell weight, and this was 4-folds increase from that of wild-type. In contrast, JH3 was a white-colored mutant that was unable to produce carotenoid pigment.

Genes involved in mating processes of saccharomyces cerevisiae (효모의 접합과정에 관여하는 유전자의 연구)

  • 장광엽;박문국;정봉우
    • Korean Journal of Microbiology
    • /
    • v.27 no.3
    • /
    • pp.210-215
    • /
    • 1989
  • In order to elucidate and characterize the signal transduction pathway(s) whereby yeast cells respond to mating pheromone, we have isolated mutants which are able to conjugate in the absence of the alpha-factor receptor. Sixty-one suppressors of a ste2-deletion mutation which also confer a ts conditional "start" arrest phenotypw have been subjected to genetic analysis. The mutants could be assigned to three complementation groups designated CDC70, CDC72 and CDC73, which are unlinked to each other as well as to the previously identified start genes. Quantitation of mating ability of the cdc70, cdc72 and cdc73 mutations in a ste2-deletion background gives levels ranging from 0.1% to 0.3% of wild type, depending on the allele and the gene. The results indicate that the signals from mating pheromone might be mediated by the CDC70, CDC72 and CDC73 products. products.

  • PDF

Potential for Efficient Synthesis of GSH Utilizing GCS1 and GLR1 Mutant Strains of Candida albicans

  • Jaeyoung SON;Min-Kyu KWAK
    • The Korean Journal of Food & Health Convergence
    • /
    • v.10 no.2
    • /
    • pp.7-11
    • /
    • 2024
  • Glutathione (GSH) is a vital compound composed of glutamic acid, cysteine, and glycine, crucial for cellular functions including oxidative stress defense and detoxification. It has widespread applications in pharmaceuticals, cosmetics, and food industries due to its antioxidant properties and immune system support. Two primary methods for GSH synthesis are enzymatic and microbial fermentation. Enzymatic synthesis is efficient but costly, while microbial fermentation, particularly using yeast strains like Candida albicans, offers a cost-effective alternative. This study focuses on genetically modifying C. albicans mutants, specifically targeting glutathione reductase (GLR1) and gamma-glutamylcysteine synthetase (GCS1) genes, integral to GSH synthesis. By optimizing these mutants, the research aims to develop a model for efficient GSH production, potentially expanding its applications in the food industry.

Genome-wide Drug-induced Haploinsufficiency Screening of Fission Yeast for Identification of Hydrazinocurcumin Targets

  • Baek, Seung-Tae;Kim, Dong-Uk;Han, Sang-Jo;Woo, Im-Sun;Nam, Mi-Young;Kim, Li-La;Heo, Kyung-Sun;Lee, Hye-Mi;Hwang, Hye-Rim;Choi, Shin-Jung;Won, Mi-Sun;Lee, Min-Ho;Park, Song-Kyu;Lee, Sung-Hou;Kwon, Ho-Jeong;Maeng, Pil-Jae;Park, Hee-Moon;Park, Young-Woo;Kim, Dong-Sup;Hoe, Kwang-Lae
    • Journal of Microbiology and Biotechnology
    • /
    • v.18 no.2
    • /
    • pp.263-269
    • /
    • 2008
  • Hydrazinocurcumin (HC), a synthetic derivative of curcumin, has been reported to inhibit angiogenesis via unknown mechanisms. Understanding the molecular mechanisms of the drug's action is important for the development of improved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screening of fission yeast gene deletion mutants has been applied to identify drug targets of HC. As a first step, the 50% inhibition concentration $(IC_{50})$ of HC was determined to be $2.2{\mu}M$. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and $4{\mu}M$. A second screening was performed to detect sensitivity to HC on the plates. The first screening revealed 178 candidates, and the second screening resulted in 13 candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficient genes gave eight target genes. Analysis of the specific targets of HC has shown that they are related to septum formation and the general transcription processes, which may be related to histone acetyltransferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay.

Role of MAPK Signaling Pathways in Regulating the Hydrophobin Cryparin in the Chestnut Blight Fungus Cryphonectria parasitica

  • So, Kum-Kang;Kim, Dae-Hyuk
    • Mycobiology
    • /
    • v.45 no.4
    • /
    • pp.362-369
    • /
    • 2017
  • We assessed the regulation of cryparin, a class II hydrophobin, using three representative mitogen-activated protein kinase (MAPK) pathways in Cryphonectria parasitica. Mutation of the CpSlt2 gene, an ortholog of yeast SLT2 in the cell wall integrity (CWI) pathway, resulted in a dramatic decrease in cryparin production. Similarly, a mutant of the CpBck1 gene, a MAP kinase kinase kinase gene in the CWI pathway, showed decreased cryparin production. Additionally, mutation of the cpmk1 gene, an ortholog of yeast HOG1, showed decreased cryparin production. However, mutation of the cpmk2 gene, an ortholog of yeast Kss1/Fus3, showed increased cryparin production. The easy-wet phenotype and accumulation of the cryparin transcript in corresponding mutants were consistent with the cryparin production results. In silico analysis of the promoter region of the cryparin gene revealed the presence of binding motifs related to downstream transcription factors of CWI, HOG1, and pheromone responsive pathways including MADS-box- and Ste12-binding domains. Real-time reverse transcriptase PCR analyses indicated that both CpRlm1, an ortholog of yeast RLM1 in the CWI pathway, and cpst12, an ortholog of yeast STE12 in the mating pathway, showed significantly reduced transcription levels in the mutant strains showing lower cryparin production in C. prasitica. However, the transcription of CpMcm1, an ortholog of yeast MCM1, did not correlate with that of the mutant strains showing downregulation of cryparin. These results indicate that three representative MAPK pathways played a role in regulating cryparin production. However, regulation varied depending on the MAPK pathways: the CWI and HOG1 pathways were stimulatory, whereas the pheromone-responsive MAPK was repressive.

Cell Cycle Regulation in the Budding Yeast

  • Nguyen, Cuong;Yoon, Chang-No;Han, Seung-Kee
    • Proceedings of the Korean Society for Bioinformatics Conference
    • /
    • 2005.09a
    • /
    • pp.278-283
    • /
    • 2005
  • Cell cycle is regulated cooperatively by several genes. The dynamic regulatory mechanism of protein interaction network of cell cycle will be presented taking the budding yeast as a sample system. Based on the mathematical model developed by Chen et at. (MBC, 11,369), at first, the dynamic role of the feedback loops is investigated. Secondly, using a bifurcation diagram, dynamic analysis of the cell cycle regulation is illustrated. The bifurcation diagram is a kind of ‘dynamic road map’ with stable and unstable solutions. On the map, a stable solution denotes a ‘road’ attracting the state and an unstable solution ‘a repelling road’ The ‘START’ transition, the initiation of the cell cycle, occurs at the point where the dynamic road changes from a fixed point to an oscillatory solution. The 'FINISH' transition, the completion of a cell cycle, is returning back to the initial state. The bifurcation analysis for the mutants could be used uncovering the role of proteins in the cell cycle regulation network.

  • PDF