Plant & Animal Genomes XI Conference

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

Poster: Large Insert Libraries
            

CONSTRUCTION OF A BAC LIBRARY FROM DIPLOID OAT (AVENA STRIGOSA)

Sainsbury Laboratory, John Innes Centre, Norwich NR4 7UH, UK

Oats synthesise antimicrobial triterpenoid saponins, that are required for disease resistance. Previously, we have generated a collection of saponin deficient (sad) mutants in the diploid oat species Avena strigosa. For further characterisation and cloning of genes involved in saponin biosynthesis, a BAC library has been constructed from A. strigosa accession S75 (1C = 3.9 pg). High molecular weight DNA was isolated from young leaf tissue. Partially digested Hind III fragments were size-selected twice and ligated into a modified pCUGI vector. Approximately 147,500 colonies (2.5% empty colonies) have been picked, stored in 384-well microtitre plates and gridded onto 8 high-density filters. The constructed library has an average insert size of ca. 110 kb, covering 4.1 haploid genome equivalents.