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Identification of immune-relevant genes from Atlantic salmon using suppression subtractive hybridization

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1007/s10126-002-0101-2
AuthorSearch for: 1; Search for: 1; Search for: 2; Search for: 2; Search for: 1; Search for: 1; Search for: 1
Affiliation
  1. National Research Council of Canada. NRC Institute for Marine Biosciences
  2. National Research Council of Canada. Human Health Therapeutics
FormatText, Article
Subjectacute-phase response (APR); Atlantic salmon; expressed sequence tag (EST); furunculosis; suppression subtractive hybridization (SSH)
Abstract
In order to probe the interaction between an invading microorganism and its host, we have investigated differential gene expression in Atlantic salmon (Salmo salar) experimentally infected with the pathogen Aeromonas salmonicida, the causative agent of furunculosis. Subtractive cDNA libraries were constructed by suppression subtractive hybridization (SSH) from 3 immune-relevant tissues at 2 time points during the infection process. Both forward- and reverse-subtracted libraries were generated, and approximately 200 clones were sequenced from each library, giving a total of 1778 expressed sequence tags (ESTs), which were annotated according to functional categories and deposited in GenBank (BQ035314–BQ037059). Numerous genes involved in signal transduction, innate immunity, and other processes have been uncovered in the subtractive libraries. These include known acute-phase reactants, along with more novel genes encoding proteins such as tachylectin, hepcidin, precerebellin-like protein, O-methyltransferase, a putative saxitoxin-binding protein, and others. A subset of genes that were represented in the subtracted libraries was further analyzed by virtual Northern, or reverse transcription–polymerase chain reaction (RT-PCR) assays to verify their differential expression as a result of infection.
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Terms of use
  • Copyright © 2004, Springer New York
In
  • Marine Biotechnology 6, no. 3 (June 2004): 199–214.
LanguageEnglish
Peer reviewedYes
NRC numberNRCC 42379
1362
NPARC number3538575
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Record identifier687661b3-2d5a-4902-b87e-ae5024ef5a40
Record created2009-03-01
Record modified2020-04-17
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