1998 Fiscal Year Final Research Report Summary
Identification and genome structure of the genes Bacteroides
| Project/Area Number |
09670283
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Wakayama Medical College |
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Principal Investigator |
AKIMOTO Shigeru Wakayama Medical College, Department of Medicine, professor., 医学部, 教授 (10159337)
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| Co-Investigator(Kenkyū-buntansha) |
OHNISHI Yoshinari Tokushima University School of Medicine, Professor, 医学部, 教授 (10037400)
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| Project Period (FY) |
1997 – 1998
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| Keywords | Bacteroides / identification / genome analysis / PFGE / phylogenetic tree / 16S-23S spacer / nucletide sequence / DNA probe |
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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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