Plant & Animal Genomes XI Conference

January 11-15, 2003
Town & Country Convention Center
San Diego, CA

Poster: Genome Sequencing & ESTs
            

FABRICATION AND APPLICATION OF A SALMONID cDNA MICROARRAY

¹ Centre for Biomedical Research, University of Victoria, Victoria, B.C., Canada, V8W 3N5

² Dept. of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, B.C., Canada, V5A 1S6

GRASP (the Genomic Research on Atlantic Salmon Project) has completed over 50,000 3’-sequences from numerous high-complexity libraries resulting in 19,000 putative transcripts. 2735 genes (2597 from Atlantic salmon, 138 from rainbow trout) were selected from 15 libraries to be PCR amplified, cleaned, and printed in duplicate onto glass slides. Several PCR products were sequenced from consolidated plates (used for printing) to ensure accurate tracking and clone identification. Two-channel hybridizations of Cy-labeled rainbow trout and Atlantic salmon brain cDNAs suggest this microarray may be used for studies involving different salmonid species. In one comparison, expression of 993 genes was detected in rainbow trout brain versus 861 genes in the Atlantic salmon brain. These gene lists had 795 genes in common and the background adjusted intensities of over 80% of these spots were less than 2-fold different between the species. Approximately 15% of the microarray genes expressed in both species’ brains, including PTEN (a tumour suppressor involved in apoptosis) and beta-2 microglobulin, had fluorescence intensities that differed by a factor greater than 2. This experiment validates the use of this microarray for cross species hybridization. This microarray is also successfully being used to profile transcripts in Atlantic salmon pre-smolt and smolt tissues and future experiments will involve embryogenesis and pathogenesis. Salmon cDNA microarrays will be valuable tools for studying salmonid development, disease, and evolution.