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

Research output: Contribution to journalArticle

Authors

  • MM Brown
  • I Katsiadaki
  • M Sanders
  • JA Craft

Colleges, School and Institutes

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.

Details

Original languageEnglish
Pages (from-to)127-130
Number of pages4
JournalMarine Environmental Research
Volume66
Issue number1
Early online date26 Feb 2008
Publication statusPublished - Jul 2008

Keywords

  • cDNA, normalisation, subtractive suppression hybridisation, microarray, stickleback