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© 1992 Oxford University Press

OTHER

Cloning of the Huntington disease region in yeast artificial chromosomes

Jian Zuo1, Carolyn Robbins1, Patricia Taillon-Miller4, David R. Cox2,3 and Richard M. Myers1,2,*

1Departments of Physiology 513 Parnassus Avenue, San Francisco, CA 94143–0444 2Biochemistry and Biophysics 513 Parnassus Avenue, San Francisco, CA 94143–0444 3Psychiatry, University of California at San Francisco 513 Parnassus Avenue, San Francisco, CA 94143–0444 4Center for Genetics in Medicine, Washington University School of Medicine Box 8232, 4566 Scott Avenue, St Louis, MO 63110, USA

*To whom correspondence should be addressed

Received May 6, 1992; Revised May 15, 1992; Accepted May 15, 1992

The gene responsible for Huntington disease has been localized to a 2.5 million base pair (Mb) region between the loci D4S10 and D4S168 on the short arm of chromosome 4. As part of a strategy to clone the HD gene on the basis of its chromosomal location, we isolated genomic DNA from the HD region as a set of overlapping yeast artificial chromosome (YAC) clones. Twenty-eight YAC clones were identified by screening human YAC libraries with twelve PCR-based sequence-tagged sites (STSs) from the region. We assembled the YAC clones into overlapping sets by hybridizing them to a large number of DNA probes from the HD region, including the STSs. In addition, we isolated the ends of the human DNA inserts of most of the YAC clones to assist in the construction of the contig. Although almost half of the YACs appear to contain chimeric inserts and several contain internal deletions or other rearrangements, we were able to obtain over 2.2 Mb of the HD region in YACs, including one continuous segment of 2.0 Mb covering the region that most likely contains the HD gene. Ten of the twenty eight YAC clones comprise a minimal set spanning the 2.2 Mb. These clones provide reagents for the complete characterization of this region of the genome and for the eventual isolation of the HD gene.


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