DNA microarrays for identifying fishes

被引：71

作者：

Kochzius, M. ^{[1
]}

Noelte, M. ^{[2
]}

Weber, H. ^{[1
]}

Silkenbeumer, N. ^{[1
]}

Hjoerleifsdottir, S. ^{[3
]}

Hreggvidsson, G. O. ^{[3
]}

Marteinsson, V. ^{[3
]}

Kappel, K. ^{[1
]}

Planes, S. ^{[4
]}

Tinti, F. ^{[5
]}

Magoulas, A. ^{[6
]}

Vazquez, E. Garcia ^{[7
]}

Turan, C. ^{[8
]}

Hervet, C. ^{[4
]}

Falgueras, D. Campo ^{[7
]}

Antoniou, A. ^{[6
]}

Landi, M. ^{[5
]}

Blohm, D. ^{[1
]}

机构：

[1] Univ Bremen, CAG, Bremen, Germany

[2] Univ Bremen, ZeTeM, Bremen, Germany

[3] Univ Perpignan, CNRS, EPHE 8046, UMR, F-66025 Perpignan, France

[4] Univ Bologna, CIRSA, I-40126 Bologna, Italy

[5] Univ Bologna, CIRSA, I-40126 Bologna, Italy

[6] Hellen Ctr Marine Res, Inst Marine Biol & Genet, Iraklion, Greece

[7] Univ Oviedo, Oviedo, Spain

[8] Mustafa Kemal Univ, Coll Fisheries & Aquaculture, Antakya, Turkey

来源：

MARINE BIOTECHNOLOGY | 2008年 / 10卷 / 02期

关键词：

DNA chips; species identification; capture oligonucleotide; pisces;

D O I：

10.1007/s10126-007-9068-3

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

In many cases marine organisms and especially their diverse developmental stages are difficult to identify by morphological characters. DNA-based identification methods offer an analytically powerful addition or even an alternative. In this study, a DNA microarray has been developed to be able to investigate its potential as a tool for the identification of fish species from European seas based on mitochondrial 16S rDNA sequences. Eleven commercially important fish species were selected for a first prototype. Oligonucleotide probes were designed based on the 16S rDNA sequences obtained from 230 individuals of 27 fish species. In addition, more than 1200 sequences of 380 species served as sequence background against which the specificity of the probes was tested in silico. Single target hybridisations with Cy5-labelled, PCR-amplified 16S rDNA fragments from each of the 11 species on microarrays containing the complete set of probes confirmed their suitability. True-positive, fluorescence signals obtained were at least one order of magnitude stronger than false-positive cross-hybridisations. Single nontarget hybridisations resulted in cross-hybridisation signals at approximately 27% of the cases tested, but all of them were at least one order of magnitude lower than true-positive signals. This study demonstrates that the 16S rDNA gene is suitable for designing oligonucleotide probes, which can be used to differentiate 11 fish species. These data are a solid basis for the second step to create a "Fish Chip" for approximately 50 fish species relevant in marine environmental and fisheries research, as well as control of fisheries products.

引用

页码：207 / 217

页数：11

共 56 条

[1] Estimating diversity of Indo-Pacific coral reef stomatopods through DNA barcoding of stomatopod larvae [J].