| Project/Area Number |
62870019
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | Kyoto University (1988)
Osaka University (1987) |
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Principal Investigator |
NISHIBUCHI Mitsuaki (1988) Associate Professor, Faculty of Medicine, Kyoto University, 医学部, 助教授 (50189304)
西淵 光昭 (1987) 大阪大学, 微生物病研究所, 助手
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| Co-Investigator(Kenkyū-buntansha) |
TAKEDA Yoshifumi Professor, Faculty of Medicine, Kyoto University, 医学部, 教授 (30029772)
MURAKAMI Akira Researcher, Central Research Laboratory, Shimazu Corporation, 技術研究本部中央研究所, 研究員
ARITA Michiko Associate Professor, Research Institute for Microbial Diseases, Osaka University, 微生物病研究所, 助手 (10127178)
HONDA Takeshi Associate Professor, Research Institute for Microbial Diseases, Osaka University, 微生物病研究所, 助教授 (60029808)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | Probe / Enterotoxigenic Escherichia coli / Heat-stable Enterotoxin / ハイブリダイゼーション |
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| Research Abstract |
Hybridization methods to detect presence or absence of the genes encoding virulence factors have been introduced recently for rapid identification of pathogenic bacteria. The methods, hoerver, have not been put to ordinary use in clinical laboratories due to technical problems. In this study, with the ultimate intention to solve the problems and develop probes of practical value, we attempted to make nonisotopically labeled synthetic oligonucleotide probes for identifying heat stable enterotixins of Escherichia coli. The results obtained are summarized as follows:
1. Hybridization tests using oligonucleotide probes representing regions of the gene encoding the heat-stable enterotoxins of E. coli revealed a region of the gene which is associated with enterotoxic activity. An oligonucleotide probe for this region detected E. coli strais producing biologically active heatstable enterotoxins.
2. We deceloped a method to label synthetic oligonucleotide probes with alkaline phosphatase via a crosslinker and spacer. The labeled oligonucleotide probes detected as low as several tens of attomoles of DNA.
3. We demonstrated that the heat stable enterotoxin-producing E. coli strains could be identified by a simple and rapid DNA colony hybridization method with a synthetic oligonucleotide probe laveled with alkaline phosphatase.
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