Plant & Animal Genomes XI Conference

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

Workshop: Comparative Genomics
            

SIMULTANEOUS HIGH-THROUGHPUT ANCHORING OF BAC-BASED PHYSICAL MAPS OF MULTIPLE SPECIES USING CORRESPONDING GENETICALLY-MAPPED STS

¹ Center for Applied Genetic Technologies, 110 Riverbend Road, University of Georgia, Athens, GA 30602, USA

² Arizona Genomics Institute, University of Arizona, Tucson, AZ 85721, USA

³ Department of Vegetable Crops, Univ. California, Davis CA 95616, USA

With many plant species entering the beginning phases of genome sequencing and BAC-based physical mapping, both comparative biology and cost-efficiency can be fostered by the use of tools that are portable to diverse taxa. We present a high-throughput approach to anchor BAC-based physical maps of multiple species, both to genetic maps and to one another. Synthetic ‘overgo’ probes were simultaneously anchored to BAC libraries of Sorghum bicolor, Sorghum propinquum, Zea mays, Oryza sativa and Saccharum (hybrid). Overgo probes were designed based on the most conserved 40 bp segment of a target sequence, as determined by cross-species hits to GENBANK (excluding repetitive DNA) and SUCEST (Private Sugarcane EST database). Overgo probes were hybridized in multiplex arrays with up to 24 probes per pool and the results deconvoluted. A total of 29 high-density BAC filters from six different libraries totaling over 500,000 BACs were tested. Hybridization and BAC fingerprint data are being used to construct physical maps using the program FPC. To date, over 4,000 probes have been tested, about half based on a 2,509 locus sorghum genetic map, and the remainder comprising BAC end sequences or specific genes of interest. The resulting physical maps can be used to transfer information from the rice sequence to other grasses. This will be demonstrated by comparison of the sorghum genetic map to the rice physical map. We will also review the feasibility of developing overgo sets that work on many diverse angiosperms, based on early results from cotton, Brassica and sorghum.