To isolate and identify new genes from Atlantic salmon using a strategy based on cloning expressed sequence tags (ESTs).
Messenger RNA (mRNA) will be prepared from nine adult salmon tissues (i.e. male and female gonad, liver, spleen, kidney, intestine, gill, brain, muscle) and used to generate cDNA libraries. Other tissues from salmon challenged with pathogens and salmon that are immuno-stimulated will also be included. Clones will be selected representing the abundant and rare mRNA transcripts in each library and 400-600 bp of their DNA sequence will be elucidated. Between 500 and 1000 ESTs will be determined from each tissue, resulting in a detailed identification of the genes that are transcriptionally active in each chosen tissue. Selected ESTs (a minimum of 50 per partner), representing the most abundant mRNAs in each respective tissue and mRNAs of particular interest based on homology searches in the international databases, will be tested for their tissue specific expression patterns among other salmon tissues.
The SALGENE partners will prepare poly(A) messenger RNA (mRNA) from the chosen salmon tissues and construct complementary DNA (cDNA) libraries using molecular genetic methods. Following this, the partners will determine partial DNA sequences of between 500 and 1000 clones from each of the eight selected salmon tissues, which will be termed expressed sequence tags (ESTs). Homology searches will be performed between the salmon ESTs and DNA sequences deposited in the international DNA databases to identify as many salmon ESTs as genes as proves possible. The SALGENE partners will also perform tissue expression analysis on selected ESTs of interest to determine tissue expression profile data.
Funding SchemeCSC - Cost-sharing contracts
FK9 4LA Stirling