PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 66pg1

THE PHYSICAL MAP OF ARABIDOPSIS THALIANA AND BEYOND.

Brian Hauge, Susan Hanley and Howard M. Goodman, Department of molecular Biology, Massachusetts General Hospital, Boston, MA 02114.


Over the course of the past several years we have assembled an overlapping cosmid map of the Arabidopsis thaliana genome, the construction of which involved the characterization from primary libraries of 20,000 clones by fingerprint analysis. Using computer matching programs the clones have been aligned into some 750 overlapping groups or contigs that encompass approximately 91,000 kb representing between 90-95% of the Arabidopsis genome. Having fingerprinted 10 genomic equivalents we have reached the practical limit of random clone mapping. The majority of the gaps in the map are expected to be small and can be attributed to the fact that the linking cosmids are either non-existent or under-represented in the cosmid libraries. To join the contigs we are selecting the linking clones from the currently available Arabidopsis YAC libraries. our linking strategy is to use YAC clones to probe ordered cosmid grids which are representative of the contigs and the unattached clones. Cosmids within a contig are chosen such that there is minimal overlap between flanking clones, yet the clones are representative of the entire contig. The representative collection of 1920 cosmids can be arrayed on two filters each the size of a microliter plate. To date we have run 226 CHEF gels, purified 2,810 random YAC clones, and completed approximately 2500 hybridizations. We are currently in the very beginning stages of comparing the hybridization linkages with the established contig map. For the physical map to be of any utility it is necessary to align it with the classical genetic linkage map. We have collaborated with Elliot Meyerowitz and Piet Stam and Maarten Koornneef in integrating the two RFLP maps and the physical and classical genetic maps of Arabidopsis. Common markers have been incorporated into the maps facilitating integration of the two data sets. The current map contains 123 classical morphological and 305 RFLP markers and a continuously updated version is available in AATDB (an Arabidopsis thaliana database).


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