Plant & Animal Genomes XIII Conference

January 15-19, 2005
Town & Country Convention Center
San Diego, CA

Transcriptome Analyses Of Rainbow Trout Oocytes

Jianbo Yao¹ , Angela D. Higgins¹ , Caird E. Rexroad²

¹  Division of Animal and Veterinary Sciences, West Virginia University, Morgantown, WV 26506

²  National Center for Cool and Cold Water Aquaculture, Kearneysville, WV 25430

Current international efforts to generate genomic resources in rainbow trout have resulted in the creation of a trout TIGR gene index containing nearly 50,000 unique expressed sequences. Although this collection of sequences represents a significant proportion of the trout genome, transcripts of oocyte/embryonic origin may be underrepresented as most of the EST sequences in this collection were derived from libraries of mixed tissue origin. Our current efforts focus on the generation of unique ESTs from unfertilized oocytes and embryos of various developmental stages. To date, we have constructed a normalized oocyte cDNA library. The normalized library was observed to have a 70-fold reduction of actin compared to the primary library as determined by southwestern ELISA blot. In addition, DNA sequence analysis of 96 clones randomly picked before and after normalization revealed that the normalization procedure successfully reduced highly abundant cDNA species. Recently, we have completed the sequencing and bioinformatic analyses of 20,000 EST sequences from this normalized library. This unique collection of EST sequences provides a valuable genomic resource for identification of genes essential for oogenesis and early embryonic development in rainbow trout.