Journal of Microbiology and Biotechnology

  • 제13권2호
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  • Pages.282-286
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  • 2003
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Cloning of a Potentially Strain-Specific DNA Probe of prevotella intermedia ATCC 25611 by Inverted Dot Blot Hybridization Screening Method

  • Kook, Joong-Ki (Department of Oral Biochemistry, Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Han, Jin-Ju (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Kim, Hwa-Sook (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Seong, Jin-Hyo (Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Kim, Dong-Kie (Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Baek, Dong-Heon (Department of Oral Microbiology and Immunology, College of Dentistry, Dankook University) ;
  • Choe, Son-Jin (Department of Oral Microbiology and Immunology, College of Dentistry, Seoul National University)
  • 발행 : 2003.04.01
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초록

The purpose of this study was to isolate a specific DNA probe for the strain ATTC 25611 of the species Prevotella intermedia by using a new rapid screening mothod. The whole-genomic DNA of P. intermedia ATCC 25611 was isolated and purified. The HindIII-digested genomic DNAs from the strain were cloned by the random cloning method. To screen the strain-specific DNA probe, inverted dot blot hybridization tests were performed. In this assay, 20 ng of recombinant plasmids containing the HindIII-digested genomic DNA fragment were boiled and blotted onto a nylon membrane, and hybridized with digoxigenin-dUTP labeled genomic DNAs in a concentration of 100 ng/ml. Southern blot analysis was performed in order to confirm the results of the inverted dot blot hybridization tests. The data showed that a Pi34 probe (2.1 kbp; 1 out of 32 probes) was specific for P. intermedia strain ATCC 25611 and could be useful for the detection and identification of the strain, particularly in epidemiological studies of periodontal disease.

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