Development and experimental validation of a 20K Atlantic cod (Gadus morhua) oligonucleotide microarray based on a collection of over 150,000 ESTs.
The collapse of Atlantic cod (Gadus morhua) wild populations strongly impacted the Atlantic cod fishery and led to the development of cod aquaculture. In order to improve aquaculture and broodstock quality, we need to gain knowledge of genes and pathways involved in Atlantic cod responses to pathogens and other stressors. The Atlantic Cod Genomics and Broodstock Development Project has generated over 150,000 expressed sequence tags from 42 cDNA libraries representing various tissues, developmental stages, and stimuli. We used this resource to develop an Atlantic cod oligonucleotide microarray containing 20,000 unique probes. Selection of sequences from the full range of cDNA libraries enables application of the microarray for a broad spectrum of Atlantic cod functional genomics studies. We included sequences that were highly abundant in suppression subtractive hybridization (SSH) libraries, which were enriched for transcripts responsive to pathogens or other stressors. These sequences represent genes that potentially play an important role in stress and/or immune responses, making the microarray particularly useful for studies of Atlantic cod gene expression responses to immune stimuli and other stressors. To demonstrate its value, we used the microarray to analyze the Atlantic cod spleen response to stimulation with formalin-killed, atypical Aeromonas salmonicida, resulting in a gene expression profile that indicates a strong innate immune response. These results were further validated by quantitative PCR analysis and comparison to results from previous analysis of an SSH library. This study shows that the Atlantic cod 20K oligonucleotide microarray is a valuable new tool for Atlantic cod functional genomics research.
Ocean Sciences Centre, Memorial University of Newfoundland, 1 Marine Lab Road, St. John's, NL A1C5S7, Canada.
Marije
M
Booman
Tudor
T
Borza
Charles Y
CY
Feng
Tiago S
TS
Hori
Brent
B
Higgins
Adrian
A
Culf
Daniel
D
Léger
Ian C
IC
Chute
Anissa
A
Belkaid
Marlies
M
Rise
A Kurt
AK
Gamperl
Sophie
S
Hubert
Jennifer
J
Kimball
Rodney J
RJ
Ouellette
Stewart C
SC
Johnson
Sharen
S
Bowman
Matthew L
ML
Rise
D048409:Aeromonas salmonicida / Q000276:immunology; D000818:Animals; D017931:DNA Primers / Q000235:genetics; D020224:Expressed Sequence Tags; D048389:Gadus morhua / Q000235:genetics* / Q000276:immunology; D020869:Gene Expression Profiling; D015723:Gene Library; D023281:Genomics; D007113:Immunity, Innate / Q000235:genetics*; D013058:Mass Spectrometry; D030381:Nodaviridae / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000276:immunology
D017931:DNA Primers; D015345:Oligonucleotide Probes