| Project/Area Number |
62560199
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General fisheries
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| Research Institution | Miyazaki University |
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Principal Investigator |
AOKI Takashi Fac. Agriculture, Miyazaki Uni. Associate professor, 農学部, 助教授 (00051805)
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| Co-Investigator(Kenkyū-buntansha) |
KITAO Tadatoshi Fac. Agriculture, Miyazaki Uni. Professor, 農学部, 教授 (60094075)
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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 |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Fish-pathogenic bacteria / Pasteurella piscicida / Vibrio anguillarum / Rapid identification / Specific gene / Probe / Vibrio anguillarum / 遺伝子構造 / 迅速同定法 |
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| Research Abstract |
Recently, randomly cloned chromosome DNA fragments and cloned gene encoded with some charcteristics have been used as specific hybridization for rapid identification of organsisms. A 692 base pair DNA fragment from fish pathogen,pasteurella piscicida, was cloned into pUC9 and as a hybridization for identification of P. piscicida. The radio labeled probe hybridized with itself and did not hybridize with other fish pathogens and closely related organsims; P. multocida, p. haemolytica, and Haemophilus influenzae. Photobiotin labeled probe hybridization detected chromosomal DNA of P. piscicide. The radio labeled probe detected P. piscicida on nitrocellulose filters which were smeared with blood and kidney tissue of yellowtail (Seriola quinqueradiata).
A 562 base pair fragment of DNA froma serotype A strain of Vibrio anguillarum was cloned into puc9 and used as a hybridization probe. The probe was tested on nine different fish pathogens, 15 Vibrio isolates, 2 organisms closely related to Vibrio, and 9 serotypes of V. anguillarum. The probe hybridized only with the DNA of V. anguillarum serotypes A and H.
The cloned specific DNA probe can be used for rapid detection and identification of fish pathogens of P. piscicida and V. anguillarum.
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