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KMID : 0545120030130020282
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Journal of Microbiology and Biotechnology
2003 Volume.13 No. 2 p.282 ~ p.286
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Cloning of a Potentially Strain-Specific DNA Probe of Prevotella intermedia ATCC 25611 by Inverted Dot Blot Hybridization Screening Method
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KOOK, JOONG-KI
HAN, JIN JU/KIM, HWA-SOOK/SEONG, JIN-HYO/KIM, DONG-KIE/BAEK, DONG-HEON/CHOE, SON-JIN
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Abstract
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The purpose of this study was to isolate a specific DNA probe for strain ATTC 25611 of the species Prevotella intermedia by using a new rapid screening method. The whole-genomic DNA of P. intermedia ATTC 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 ATTS 25611 and be useful for the detection and identification of the strain, particularly in epidemiological studies of periodontal disease.
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KEYWORD
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