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© 1993 Oxford University Press

RESEARCH-ARTICLE

The isolation of a yeast artificial chromosome (YAC) contig extending for 2 megabases in the vicinity of the Von Hippel Lindau disease gene

Wanguo Liu, Marie Plechockl, Viji Shridhar, Gregory Lyles, Zheng Song, Yasuke Nakamura1, Harry Drabkin2, Jeffrey Vance3 and David I. Smith*

Molecular Biology and Genetics, Wayne State University School of Medicine 540 E Canfield, Detroit, MI 48201, USA 1Division of Biochemistry, Cancer Institute Tokyo 170, Japan 2University of Colorado Health Sciences Center Denver, CO 82006 3Division of Neurology, Duke University Medical Center Durham, NC 27710, USA

* To whom correspondence should be addressed

Received April 20, 1993; Revised May 20, 1993; Accepted May 20, 1993

Von Hippel LIndau disease (VHL) is a rare autosomal dominant disease associated with tumors and cysts in multiple organ systems. The VHL disease gene is tightly linked to the polymorphic DNA marker 233E2 (D3S720) and flanked by 479H4 (D3S719) on its telomeric and RAF1 on its centromeric side. Two additional markers, D3S1038 and D3S601, have also been identified, and these markers, like D3S720, are very tightly linked to VHL. Previously 93 cosmid clones were mapped to the larger region, 3p24.2 - pter, surrounding the VHL disease gene (1). Using a Southern-based screening strategy on pools of YAC clones we have Isolated a contig of overlapping YAC clones that extends about 0.7 megabase centromeric, and about 1.3 megabases telomeric of D3S720 and contains all three tightly linked VHL markers. Individual YACs In this contig were hybridized to grids containing cosmids localized between 3p24.2-pter and to several cosmids localized by fluorescent in situ hybridization (FISH) to 3p25. A total of 28 cosmids were positioned on this contig of overlapping YAC clones. We have also identified homologous YAC clones to many additional cosmid clones localized between 3p24.2–p25, although these have not yet been precisely localized relative to the contig of YAC clones. This contig of YAC clones probably contains the VHL disease gene and should facilitate the isolation and characterization of this gene.


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