• Title/Summary/Keyword: mutant strain

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Analysis of Antifungal Proteins in Streptomyces sp. SAR01 (Streptomyces sp. SAR01 균주에서의 항진균 관련 단백질 분석)

  • 이영근;김재성;조규성;장병일;추철형
    • Korean Journal of Environmental Biology
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    • v.20 no.3
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    • pp.237-244
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    • 2002
  • To analyze proteins related to antifungal activity, SAR01 strain was isolated from seaweed and identified as Streptomyces sp. from the result of FAME (fatty acid methyl ester) analysis. The isolated strain had antifungal activities against T species of plant pathogenic fungi. Antifungal activity deficient mutant (SAR 535) of Streptomyces sp. SAR01 was induced by gamma radiation $(^{60}Co,\;5kGy)$. By 2 D electrophoresis analysis, 6 protein spots were found in wild strain (SAR01) but these spots disappeared in mutant strain (SAR535). Among them, 5 proteins showed similarities to heat shock protein 70(HSP70), Fe-containing superoxide dismutase II (Fe- SODII), ribosome recycling factor (RRF), 10 kDa chnperonin (GroES) and inorganic pyrophosphatase (PPAse), respectively. It suggested that the above 6 proteins could be closely related to the antifungal activity of Streptomyces sp. SAR01.

Identification of Fluorescent Pseudomonads Producing Siderophore and Construction of Siderophore Biosynthesis Defective Mutant (Siderophore를 생성하는 Fluorescent Pseudomonads의 분리, 동정 및 돌연번이 유기)

  • Park, Yeal;Kim, Hyun Hee;Myeong-gu Yeo;Young-woo Seo;Han-cheol Koh;Young-gi Yang;Hyeon-Sook Cheong;Sung-jun Kim
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.286-290
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    • 1992
  • The present study was performed to isolate the fluorescent pseudomonads from Kwang-Ju soil and to construct a mutant strain defective in siderophore biosynthesis. The siderophore-secreting pseudomonads were screened on Blue agar (Chrome Azuol S agar) plates and one strain of them was designated to Pseudominas fluorescens (P. fluorescens) PY002. To construct a mutant defective in siderophore biosynthesis, P. fluorescens PY002 was randomly mutagenized with a transposon Tn5. The location of Tn5 integrated into chromosomal of the mutants strain was determined by Southern blot analysis. The mutagenized strain showed non-fluorescent on a King's B agar plate and were defective in iron (III) acquisition ability.

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Electrophoretic Analysis of Total Proteins in Bacillus sphaericus ts-Dl290 (Bacillus sphaericus ts-D1200 단백질의 전기영동적 분석)

  • 서정희;이형환;김영희
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.541-546
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    • 1990
  • Bacillus sphaericus ts-Dl290 was characterized by SDS-PAGE produced by the mutant at $30^{\circ}C$ and $42^{\circ}C$. The total amount of proteins produced by the mutant at $42^{\circ}C$ decreased to one-fifth of those at $30^{\circ}C$; however, when the culture was shifted down from $42^{\circ}C$ after 4 to $30^{\circ}C$, the total amount of protein decreased to one-third and the 221 kd protein did not appear, but the 155 kd appeared remarkably. When the mutant and the wild type strain were cultured in the media containing 80$\mu g$ per ml of chloramphenicol at $42^{\circ}C$, the wild type strain synthesized half amounts of the total proteins than those at $30^{\circ}C$, and the mutant produced one-tenth of the total protein amounts. When the both strains were cultured in the media containing chloramphenicol, the 155 kd protein was produced was produced in lesser amounts than those without chloramphenicol. The 150 kd protein showed lethal activity to Culex pipiens 3rd instar larvae.

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Production of Glutathione by the Yeast Mutant Saccharomyces cerevisiae Sa59 (효모변이주 Saccharomyces cerevisiae Sa59에 의한 glutathione 생성)

  • Jang, Hye-Yoon;Oh, Chul-Hwan;Oh, Nam-Soon
    • Korean Journal of Food Science and Technology
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    • v.45 no.6
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    • pp.801-804
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    • 2013
  • The glutathione contents of the selected mutants were investigated and found to be 6.1-15.8 mg/g-DCW. The glutathione content positively correlated with the antioxidant activity of the mutant strains ($R^2$=0.488). Furthermore, the glutathione content of the mutant S. cerevisiae Sa-59 was approximately 38% greater than that of the wild type strain and, therefore, this mutant strain was selected for glutathione production. The volumetric glutathione content in a shaking culture was increased by about 70% compared to the static culture. In addition, the specific glutathione content was increased by ~19%. The volumetric glutathione content and specific glutathione content were increased by approximately 16% and 66%, respectively, when 0.04% glutamate, 0.04% cysteine and 0.04% glycine were added. Furthermore, the highest antioxidant activity was 0.52 as absorbance unit at 700 nm.

Improved Plant Growth from Seed Bacterization Using Siderophore Overproducing Cold Resistant Mutant of Pseudomonas fluorescens

  • Katiyar, Vandana;Goel, Reeta
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.653-657
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    • 2004
  • The cold resistant mutants of P. fluorescens strain $PRS_{9}$ and ATCC13525 were developed which could grow equally well at $28^{\circ}C$ and $10^{\circ}C$. All the mutants were tested for siderophore production, of which $CRPF_9$ (ATCC13525 mutant) was selected, as there was a 16.8-fold increase when compared to its wild-type. Under in vitro conditions, $CRPF_9$ showed better growth promotion both in wheat (29.1% increase in root length) and mung bean (51.5% increase in root length) at $10^{\circ}C$. Greenhouse trials showed a significant increase in root (13.84cm) and shoot (15.0cm) length of $CRPF_9$-treated mung bean seeds, indicating increased rhizocompetence of the mutant. Ferric citrate was a better iron source than ferric hydroxide for plant growth.

Energy Status of Neurospora crassa Mutant nap in Relation to Accumulation of Carotenoids

  • Belozersk, Tatyana A.;Potapova, Tatyana V.;Isakova, Elena P.;Shurubor, Eugene I.;Savel'eva, Ludmila V.;Zvyagilskaya, Renata A.
    • Journal of Microbiology
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    • v.41 no.1
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    • pp.41-45
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    • 2003
  • N crassa mutant strain nap showed reduced growth rate, decreased electric membrane potential, and elevated intracellular ATP content in comparison to the wild type. Blue light induced a hyperpolarization of the membrane potential in both strains. The analysis of oxidative and phosphorylation activities of mitochondria isolated from the two strains has revealed that nap utilized more efficient oxidative pathways. The higher intracellular ATP content in the nap was presumably due to impaired transport systems of the plasma membrane, and to a lesser extent to the functioning of the fully competent respiratory chain. The excess ATP possibly accounts for carotenoid accumulation in the mutant.

Isolation of Oligosporogenous (Spo-Cry+)Mutant Containing Relatively tow Hemolytic Activity in $\delta$-Endotoxin from Bacillus thuringiensig subsp, israelensis (Bacillus thuringiensig subsp, israelensis의 용혈성 약화 및 무포자 변이주의 분리)

  • 배점순;김광현
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.273-276
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    • 1989
  • The isolate SPO 3, an oligosporogenous and crystalliferous mutant, which was isolated from Bacillus thuringiensig subsp, israelensis by heat treatment at 42$^{\circ}C$ for 24 hit. The $\delta$-endotoxin of the mutant had a relatively low hemolytic activity on human red blood cells; the $\delta$-endotoxin of the mutant had 25 times less hemolytic activity compared to that of wild strain. The loss of 28 KDa hemolytic protein subunit in $\delta$-endotoxin of the mutant was confirmed by means of double immunodiffusiori, immunoelectrophoresis, and SDS-PAGE.

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Generation of an Arginine Auxotrophic Mutant of Colletotrichum acutatum as a Recipient Host for Insertional Mutagenesis

  • Kim, Hee-Kyoung;Lee, Sun-Hee;Kim, Heung-Tae;Yun, Sung-Hwan
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.205-212
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    • 2009
  • Colletotrichum acutatum was the main cause of the recent outbreaks of anthracnose on pepper fruit in Korea. To facilitate molecular analysis of C. acutatum, we generated an arginine auxotrophic mutant of the C acutatum strain JC24 using a targeted gene replacement strategy. A 3.3-kb genomic region carrying an ortholog (designated CaARG2) of the fungal gene encoding N-acetylglutamate synthase, the first enzyme of arginine biosynthesis in fungi, was deleted from the fungal genome. The mutant exhibited normal growth only when arginine was exogenously supplied into the culture medium. Transformation of the arginine auxotrophic mutant with a plasmid DNA carrying an intact copy of CaARG2, which was smaller than the deleted region in the mutant, not only caused random vector insertions in the fungal genome, but also recovered both hyphal growth and pathogenicity of the mutant to the wild-type level. Using this new selection system, we have successfully developed a restriction enzyme-mediated integration procedure, which would provide an economically efficient random mutagenesis method in C. acutatum.

Proteomic Analysis of the GacA Response Regulator in Pseudomonas chlororaphis O6

  • Anderson, Anne J.;Kim, Young Cheol
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.162-169
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    • 2018
  • The GacS/GacA system in the root colonizer Pseudomonas chlororaphis O6 is a key regulatory system of many traits relevant to the plant probiotic nature of this bacterium. The work in this paper elucidates proteins using proteomics approach in P. chlororaphis O6 under the control of the cytoplasmic regulatory protein, GacA. A gacA mutant of P. chlororaphis O6 showed loss in production of phenazines, acyl homoserine lactones, hydrogen cyanide, and protease, changes that were associated with reduced in vitro antifungal activity against plant fungal pathogens. Production of iron-chelating siderophore was significantly enhanced in the gacA mutant, also paralleling changes in a gacS mutant. However, proteomic analysis revealed proteins (13 downregulated and 7 upregulated proteins in the mutant compared to parental strain) under GacA control that were not apparent by a proteomic study of a gacS mutant. The putative identity of the downregulated proteins suggested that a gacA mutant would have altered transport potentials. Notable would be a predicted loss of type-VI secretion and PEP-dependent transport. Study of mutants of these GacA-regulated proteins will indicate further the features required for probiotic potential in this rhizobacterium.

Survival Factor Gene FgSvf1 Is Required for Normal Growth and Stress Resistance in Fusarium graminearum

  • Li, Taiying;Jung, Boknam;Park, Sook-Young;Lee, Jungkwan
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.393-405
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    • 2019
  • Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.