Plant & Animal Genomes XI Conference Plant & Animal Genomes XI ConferenceJanuary 11-15, 2003
Town & Country Convention Center
San Diego, CA
Poster: Genome Sequencing & ESTs
A cDNA library was constructed from brain stem, the lateral aspect of the cerebellum, frontal cortex and the hypothalamus, and 215,000 clones spotted at high-density on to 8 nylon filters. The filters were probed with 220 x 8-mer oligo-nucleotides and hybridisation patterns analysed. The oligo-nucleotide fingerprints produced were compared, and clones with similar patterns were placed into clusters. From the 215,000 clones, 12,432 formed clusters and 49,461 remained as singleton clones. A first approximate identification of the genes represented by the clones was achieved by producing oligo-nucleotide fingerprints in silico for human and bovine Unigene sets, and comparing these fingerprints with those obtained for the cDNA clones. Some redundancy was found among the 12,432 clusters so representative clones from 8,919 clusters and additionally 14,121 singleton clones were selected to make up a non-redundant set of clones to produce expression arrays. The identities of these clones are now being verified by sequencing. A macro-array of ~23,000 clones printed onto nylon membranes has been tested by hybridisation against several bovine tissues. Both the macro-arrays on nylon and micro-arrays on glass will be available from the resource centres: RZPD in Berlin and ARK Genomics in Roslin. Further work will identify additional cDNAs from immune tissues and the gut, that are not represented in the brain set, by complex hybridisations of probes made from the non-redundant set against cDNAs from these other tissues. Novel cDNA clones identified will be sequenced and added to the array set to create a comprehensive bovine expression array.