Analysis of gene structure and development of rapid identification of fish pathogens

鱼类病原体基因结构分析及快速鉴定技术开发

• 批准号: 62560199
• 负责人: AOKI Takashi
• 金额: $ 1.34万
• 依托单位: Miyazaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560199/
• 关键词: Fish-pathogenic bacteria; Pasteurella piscicida; Vibrio anguillarum; Rapid identification; Specific gene; Probe; Vibrio anguillarum; 遺伝子構造; 迅速同定法

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.

近年来，随机克隆的染色体DNA片段和编码有一定特征的克隆基因已被用于生物的特异性杂交。将鱼源piscicpasteurella piscicida 692碱基对DNA片段克隆到pUC9中，并进行杂交鉴定。放射性标记探针与自身杂交，不与其他鱼类病原体和密切相关的生物体杂交；多杀性假单胞菌、溶血假单胞菌和流感嗜血杆菌。光生物素标记探针杂交检测杀鱼鱼染色体DNA。用放射性标记探针在涂有黄尾鱼血液和肾组织的硝化纤维素滤料上检测到鱼酸假单胞菌。将鳗弧菌A血清型菌株的562碱基对DNA片段克隆到puc9中作为杂交探针。该探针对9种不同的鱼类病原体、15株弧菌、2株与弧菌密切相关的生物以及9种鳗弧菌血清型进行了检测。该探针仅与A、h两种血清型的鱼源DNA进行杂交，克隆出的特异性DNA探针可用于鱼源疫病和鳗源疫病的快速检测和鉴定。

项目成果

I.Hirono.;T.Aoki.: Proceedings of 5th International symposium on Microbial Ecology.

I.Hirono.；T.Aoki.：第五届国际微生物生态学研讨会论文集。

Zhao, T.;T. Aoki and T. Kitao: 日本水産学会誌.

赵，T.；T. Aoki 和 T. Kitao：日本水产科学学会杂志。

Jiang Z.;T.Aoki: Diseases of Aquatic Organisms.

Jiang Z.；T.Aoki：水生生物疾病。