• Title/Summary/Keyword: PEGE

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Separation of large DNA molecules by pulsed field gel electrophoresis (역전장 전기영동장치를 이용한 대형 DNA 분리에 관한 연구)

  • Joo, Yi-seok;Casey, Thomas A;Yoon, Yong-dhuk
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.81-85
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    • 1993
  • Gel electrophoresis has proven to be one of the most useful of DNA separation and purification. The new technique of pulsed field gel electrophoresis (PFGE) is high resolution separation of large size DNA moleculs. Conventional continuous gel electrophresis can not be separation of large DNA fragments(20~50 k base). Field inversion gel electrophoresis(FIGE) is very useful for large DNA molecules. We have found that a pulse ratio ; 2 : 1, time ; 24hrs., volts ; $10^{volts}/_{cm}$, start ; 0.45sec, end ; 1sec, is most effectively resolves DNA fragment in the 6~50k base.

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Megabase-sized DNA isolation and electrophoretic karyotype of fusarium oxysporum schlecht

  • Park, Min-Seon;Min, Byung-Re
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.132-135
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    • 1995
  • To investigate the electrophoretic karytype of Fusarium oxysporum, intact chromosomal DNA was separated by pulsed-field gel electrophoresis (PEGE). DNA extraction from nulcei, mycelia and protoplasts were compared with one another and with the quantity and the suitability for PFGE separation in agarose gel. As a result, the most useful extracting method for intact DNA was found to be that from protoplasts. By varying the electrophoretic conditions, 8 chromosomal DNA bounds were resolved. Using the Schizosaccharomyces pombe and Saccharomyces cerevisiae as size standards, the size of Fusarium oxysporum chromosomes was estimated to range from approximately 0.6 Mb TO 6.7 Mb, and total genome size was 26.7 Mb. The suitability of electrophoretic karyotyping as a tool for strain characterization is discussed.

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Structural Analysis of Class I Integron Gene Cassette and Assessment of Genetic Relationships by PFGE of Salmonella enterica Serovar Typhimurium Isolated in Gyeongbuk Area (경북지역에서 분리된 Salmonella enterica Serovar Typhimurium의 Class I Integron Gene Cassette 특성과 PEGE 유형분석)

  • Sohn, Chang-Kyu;Lee, Jung-A;Lee, Do-Young;Hun, Wan;Jung, Jung-Kyo
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.12-18
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    • 2006
  • Thirty five Salmonella enterica serovar Typhimurium strains were isolated from diarrheic patients and pigs in Gyeongbuk area from 2003 to 2004. All 35 strains (17 strains from diarrheic patients and 18 from pigs) were resistant to more than one drug and most of strains isolated from pigs were resistant to ampicillin, ohloram-phenicol, streptomycin, sulfamethoxazole-trimethoprime, tetracyclin and nalidixic acid. Each isolate was also screened or the presence of class I, II and III integron gene cassettes. Among 35 strains,3 out of 17 strains isolated from diarrheic patients, carried dhfrX-orfF-aadA2 integron gene cassette and among 18 strains isolated from diseased pigs, 11 strains carried dhfrX-orfF-aadA2 integron gene cassette and 1 strain carried aadA2 integron only. But any class II and class II integron gene cassette were not detected in 35 strains. Thirty five strains were divided by five pulsotypes. Thirty one strains out of thirty five were pulsotype A. Among the remaining 4 strains, one each strain belonged to pulsotype B, C, D and pulsotype E. This data of pulsotypes showed that the widespread of pulsotype A, Salmonella enterica serovar Typhimurium in human and pigs in Gyeongbuk area may have been caused by the dissemination of a few epidemic strains in this area. Thirteen strains contain dhfrX-orfF-aadA2 integron gene cassette showed pulsotype A and one strain contains dhfrX-orfF-aadA2 integron gene cassette showed pulsotype B. One strain contains aadA2 integron showed pulsotype E. But fifteen strains do not contain any integron showed pulsotype A.