Plant, Animal & Microbe Genomes X Conference Plant, Animal & Microbe Genomes X ConferenceJanuary 12-16, 2002
Town & Country Convention Center
San Diego, CA
Poster: Large Insert Libraries
A bacterial artificial chromosome (BAC) library was constructed using nuclear DNA from homozygous male asparagus plants to provide a ressource for map-based cloning of the sex determining gene. The basis for a map-based cloning strategy is a high-resolution genetic map in the region of the target gene. We are currently developing a genetic map based on 924 individuals of five populations of selfed andromonoecious plants. Three AFLP and four sequence-tagged site (STS) markers closely linked to the sex-locus were identified. These markers linked in a distance between 0 cM and 0.9 cM are being used to screen the BAC-library. The library contains a total of 79,488 clones from three separate ligation reactions with an average insert size of 85 kb, representing approximately 5 haploid genome equivalents, excluding false positive clones. The frequency of BAC clones carrying inserts of chloroplast (cp) and mitochondrial (mt) DNA was estimated to be 0.6 % and 2.0 %, respectively. To screen the BAC-library a pooling strategy was followed in which one pool was formed by four 96-well microtitre plates. The identification of sex-linked clones and the development of BAC contigs to cover the sex gene region is underway.