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Abstract
The three-spined stickleback (Gasterosteus aculeatus) is ideally suited to laboratory studies, while its wide distribution in the northern hemisphere gives it great potential as a sentinel organism. In the setting of a UK-wide collaboration (Fish Toxicogenomics) we have developed a microarray for transcriptomic analysis of chemical responses in populations of G. aculeatus under laboratory and field conditions. Although several EST libraries are available for this species none are from chem i cal -exposed fish and thus unlikely to include a full set of potlutant-responsive genes. To harvest such transcripts cDNA libraries were produced from liver of chemical-exposed mature males. Two normalised full-length libraries were generated by different methods: (1) partial subtraction of polyA' RNA against solid-phase cDNA using magnetic bead technology; (2) degradation of double stranded cDNA formed by abundant transcripts. To enrich for pollutant-responsive genes a subtracted EST library was also generated. For each library -1.5 K clones were sequenced and characterised using Blast2GO. All libraries contained pollutant-responsive transcripts not Previously available while additionally the subtracted library was generally enriched -1.210-fold for transcripts expected to be induced in response to the pollutants. (C) 2008 Elsevier Ltd. All rights reserved.
Original language | English |
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Pages (from-to) | 127-130 |
Number of pages | 4 |
Journal | Marine Environmental Research |
Volume | 66 |
Issue number | 1 |
Early online date | 26 Feb 2008 |
DOIs | |
Publication status | Published - Jul 2008 |
Keywords
- cDNA
- normalisation
- subtractive suppression hybridisation
- microarray
- stickleback
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Dive into the research topics of 'Construction of subtracted EST and normalised cDNA libraries from liver of chemical-exposed three-spined stickleback (Gasterosteus aculeatus) containing pollutant-responsive genes as a resource for transcriptome analysis'. Together they form a unique fingerprint.Projects
- 1 Finished
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Identifying and Defining the Bases of Individual and Population Susceptibility and Adaption to Environmental Pollutants in Fish: An Integrated Transcriptomic and Metabolomic Approach
Chipman, K., Falciani, F., Viant, M. & Minchin, S.
Natural Environment Research Council
1/04/05 → 30/11/08
Project: Research Councils