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学科主题: Oceanography
题名: Identification of Prorocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry
作者: Hou Jianjun1, 2, 3, 4, 5; Lai Hongyan3, 4; Huang Bangqin1; Chen Jixin1
通讯作者: Huang, BQ, Xiamen Univ, Environm Sci Res Ctr, State Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China
关键词: oligonucleotide ; DNA probes ; Prorocentrum minimum ; Takayama pulchella ; fluorescence in situ hybridization ; flow cytometry
刊名: ACTA OCEANOLOGICA SINICA
发表日期: 2009
卷: 28, 期:2, 页:103-114
收录类别: SCI
文章类型: Article
部门归属: [Hou Jianjun; Huang Bangqin; Chen Jixin] Xiamen Univ, Environm Sci Res Ctr, State Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China; [Hou Jianjun] Huazhong Agr Univ, Coll Fisheries, Wuhan 430070, Peoples R China; [Hou Jianjun; Lai Hongyan] Hubei Normal Univ, Coll Life Sci, Huangshi 435002, Peoples R China; [Hou Jianjun; Lai Hongyan] Yangtse River Fisheries Res Inst, Key Lab Freshwater Fish Germplasm Resources & Bio, Jinzhou 434000, Peoples R China; [Hou Jianjun] Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China
WOS标题词: Science & Technology ; Physical Sciences
资助者: Fujian Provincial Government of China [2005YZ1018]; Xiamen Municipal Government of China [3502220041059]; China Postdoctoral Science Foundation [20060400854]; Open Fund of the State Key Laboratory of Freshwater Ecology and Biotechnology ; Institute of Hydrobiology, Chinese Academy of Sciences [2008FB005]; Specialized Research Fund for the Doctoral Program of Higher Education of China [20070504076]; Open Fund of the Key Laboratory of Freshwater Fish ; Key Laboratory of Freshwater Fish Germplasm and Biotechnology of Ministry of Agriculture ; Chinese Academy of Fishery Sciences [LFB20070617]; National Natural Science Foundation of China [40576055]
类目[WOS]: Oceanography
研究领域[WOS]: Oceanography
摘要: Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced. and these sequence data were deposited in the GenBank. Eight oligonucleotide probes (DNA probes) were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order: PM18S02 > PM28S02 > PM28S01 > PM18S01, and that of the probes specific to T. pulchella was TP18S02 > TP28S01 > TP28S02 > TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02; TP18S02 and TP28S01, and the protocol, were also useful for the detection of algae in natural samples.
英文摘要: Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced. and these sequence data were deposited in the GenBank. Eight oligonucleotide probes (DNA probes) were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order: PM18S02 > PM28S02 > PM28S01 > PM18S01, and that of the probes specific to T. pulchella was TP18S02 > TP28S01 > TP28S02 > TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02; TP18S02 and TP28S01, and the protocol, were also useful for the detection of algae in natural samples.
关键词[WOS]: TYRAMIDE SIGNAL AMPLIFICATION ; WHOLE-CELL HYBRIDIZATION ; TOXIC DINOFLAGELLATE ; ALEXANDRIUM-TAMARENSE ; RNA-CONTENT ; DINOPHYCEAE ; PROBES ; OLIGONUCLEOTIDE ; VARIABILITY ; CATENELLA
语种: 英语
WOS记录号: WOS:000266647100011
ISSN号: 0253-505X
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/7726
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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作者单位: 1.Xiamen Univ, Environm Sci Res Ctr, State Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China
2.Huazhong Agr Univ, Coll Fisheries, Wuhan 430070, Peoples R China
3.Hubei Normal Univ, Coll Life Sci, Huangshi 435002, Peoples R China
4.Yangtse River Fisheries Res Inst, Key Lab Freshwater Fish Germplasm Resources & Bio, Jinzhou 434000, Peoples R China
5.Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan 430072, Peoples R China

Recommended Citation:
Hou Jianjun; Lai Hongyan; Huang Bangqin; Chen Jixin.Identification of Prorocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry,ACTA OCEANOLOGICA SINICA,2009,28(2):103-114
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