Application of In Vivo Mapping Strategies to Order Arabidopsis YAC Libraries

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The objective of this award is the isolation, identification, and mapping of large segments of the Arabidopsis genome using yeast artificial chromosome (YAC) vectors and the construction of new Arabidopsis YAC libraries. This research takes advantage of newly developed pulse field gel sizing methods and yeast transformation approaches to enrich for Arabidopsis YAC clones that contain larger average inserts than are present in the existing libraries. The new, larger libraries should reduce the number of steps in a chromosome walking experiment by greater than a factor of two, thus facilitating the identification of specific genes of interest. The utility of the research lies in rapid distribution and dissemination of the library to the entire scientific community. Drs. Chory and Hoekstra will establish methods for clone archiving and data storage to catalog the library, and to place existing markers onto known clones in collaboration with the Howard Goodman's group at Massachusetts General Hospital. Copies of the YAC library will be distributed to interested researchers through the Arabidopsis Biological Resource Center at Ohio State University.