• Title/Summary/Keyword: CHEF-PFGE

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Study of Electrophoretic Karyotypes of Fusarium Section Liseola (Fusarium 균의 section Liseola에 대한 핵형 연구)

  • 밍병례;안미선;최영길
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.192-196
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    • 1999
  • CHEF-PFGE(Contour-Clarnped Homogeneous Electric field- Pulsed Field Gel Electrophoresis) was used to identify electrophoretic karyotype for eight strains belonging to the Fzisoriuni section Liseolo. Chromosome numbers were nine to thirteen bands, ranging in size Cram 0.75 to 6.45 Mb. The total genome size was eslimated to range from 38.19 Mb to 43.12 Mb and numerous chromosome-length polymorphisms (CLPs) were observed. For the chromosome localizalion of the gene, 1GS sequence(2.6 Kb) of rDNA from F: moniliforme, chs-2 gene(2.8 Kb) and 4 - 3 gene(3.8 Kb) from Neuuospora cmssa were wed as probes.

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Comparison of Biotyping, Serotyping and Molecular Typing of Yersinia enterocolitica Isolated from Spring water in Seoul (서울시내 약수에서 분리한 Yersinia enterocolitica의 생물형, 혈청형 및 분자학적 형별비교)

  • 이영기;최성민;오수경;신재영
    • Journal of environmental and Sanitary engineering
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    • v.14 no.4
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    • pp.99-109
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    • 1999
  • Enteropathogenic Yersina enterocolitica is an important cause of human and animal disease. Phenotypic and genotypic characteristics currently used to identify Yersinia enterocolitica are not necessarily sufficient to differentiate pathogenic from non-pathogenic strains or to analyze the epidemiology of yersiniae at a molecular level. To improve the characterization of Yersinia enterocolitica, A total of 65 isolates of Yersinia enterocolitica were examined with bioserotyping, antibiotic susceptibilities, PFGE, PCR-ribotyping. Genomic DNA pattern generated by PFGE are highly specific for different strains of an organism and have significant value in epidemiologic investigations. The PFGE analysis of Not I-digested chromosomal DNA of Y. enterocolitica were performed with a CHEF Mapper(Bio-Rad, USA). Not I generated 19 restriction endonuclease digestion profiles(REDP). PCR-ribotyping, performed with primers complementry to conserved regions of 16S and 23S rRNA gene, generated 13 ribotypes. PCR-ribotyping can be considered a good technich for subtyping strains of Y.enterocolitica.

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Electrophoretic Karyotyping by PFGE in the Genus Fusarium (Fusarium속에서 PFGE를 이용한 Electrophoretic Karyotyping)

  • Min, Byung-Re;Jung, Jin-Sook;Choi, Yong-Keel
    • The Korean Journal of Mycology
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    • v.26 no.2 s.85
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    • pp.135-143
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    • 1998
  • Contour-clamped homogeneous electric field gel electrophoresis was used to establish electrophoretic karyotype for 10 species of Fusarium sections Sporotrichiella, Liseola, Gibbosum, Discolor and Martiella. Intact chromosomal DNA was isolated from fungal protoplast and separated under various conditions according to their size in order to improve DNA separation. The numbers of chromosome-sized DNA molecules for individual species ranged from 5-13, with individual chromosomes ranging from 0.78 Mb to 7.20 Mb in size. The total genome DNA size of each species was estimated at about 18.32 Mb to 48.20 Mb. Comparison of karyotype profiles following Southern hybridization analysis with a randomly selected genomic probe of F. oxysporum formae speciales litii was carried out.

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Electrophoretic Karyotypes of Fusarium oxysporum f. sp. lycopersici (Fusarium oxysporum f. sp. lycopersici의 Electrophoretic Karyotype)

  • Kim, Young-Tae;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.112-118
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    • 1999
  • Strains of Fusarium oxysporum f. sp. lycopersici isolated from Korea, Japan and U.S.A. were used for electrophoretic karyotype (EK) analysis. Chromosome separations on FastLane agarose gels (FMC BioProducts, Rockland, ME), called pulsed field gel electrophoresis (PFGE), were performed by CHEF-DRII apparatus (Bio-Rad Laboratories, Melville, NY) using TAE as a running buffer. To obtain optimal condition for separation of chromosome sized DNAs, variable running conditions such as field strengths, swithching intervals, and running time were applied in CHEF gel electrophoresis. We were able to resolve 9 to 11 chromosome sized DNAs ranging in size from 0.76 to 6.41 Mb in isolates from Korea and estimate that the total genome size was ranging from 35.29 to 38.92 Mb. Distinct differences in length range and genome size exist among isolates from different countries. Isolates from Japan and U.S.A. were resolved 9 to 11 chromosome sized DNAs ranging in size from 1.24 to 6.85 Mb and estimated that the total genome size was ranging from 35.32 to 43.87 Mb. Isolates from variable provinces in Korea had the same or similar chromosomal polymorphism and showed different chromosomal DNA patterns compared to isolates from the other countries.

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