| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Research Abstract |
The Chromosomal DNAs of seven Bacteroides species were digested with restriction enzyme Ceul and analysed by pulsed-field gel electrophoresis. The genome sizes of B.fragilis, B.distasonis, B.eggerthil, B.avatus, B.thetalotaomicron, B.uniformis, and B.vulgatus were determined to be 5.3, 4.8, 4.4, 6.9, 4.8, 4.6, and 5.1 Mbp, respectively. Ceul cut B.uniformis and B.eggerthii into four, B.ova tus into five, B.fragilis and B.thetaiotaomicron into six, and B.distasonis and B.vulgatus into seven fragments. On the basis of genome size, restriction profile, and Ceul fragment number, a phylogenetic tree of the Bacteroides species was proposed. This was in overall agreement with the previous phylogenetic tree obtained by 16S rRNA data, withe the exceptions of B.distasonis and B.ovatus.
Amplification of the 16S-23S spacer region was performed with seven Bacteroides species by using oligonucleotide primers complementary to the conserved region of the 16S and 23S ribosomal RNA gene. Seven Bacteroides species showed different PCR profiles which were more easily distinguished by RFLP.Characterization of sixty-three strains of B.fragilis revealed four types of PCR profiles. These genetic heterogeneity of 16S-23S spacer region in Bacteroides species can be applied to species level identification and intrasp ecies ribotyping of Bacteroides.
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