Sequence-Ready Contig for the 1.4-cM Ductal Carcinoma in Situ Lossof Heterozygosity Region on Chromosome 8p22–p23 J. C. Wang,* ,1 D. M. Radford,* ,2 M. S. Holt,* ,3 C. Helms,* ,3 A. Goate,† , ‡ ,4 W. Brandt,* 5 M. Parik,* N. J. Phillips,§ K. DeSchryver, ¶ M. E. Schuh, K. L. Fair,* J. H. Ritter,** P. Marshall,* and H. Donis-Keller,* , † , ‡ * Department of Surgery, Division of Human Molecular Genetics, †Department of Genetics, ‡Department of Psychiatry, and ** Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110; §Department of Pathology, St. Louis University School of Medicine, St. Louis, Missouri 63110; ¶ West Central Pathologists Inc., St. Louis, Missouri 63141; and Parkcrest Surgical Associates Inc., St. Louis, Missouri 63141 Received April 28, 1999; accepted June 15, 1999 We report the construction of an 1.7-Mb se- quence-ready YAC/BAC clone contig of 8p22–p23. This chromosomal region has been associated with frequent loss of heterozygosity (LOH) in breast, ovarian, prostate, head and neck, and liver cancer. We first constructed a meiotic linkage map for 8p to resolve previously reported conflicting map orders from the literature. The target region containing a putative tumor suppressor gene was defined by al- lelotyping 65 cases of sporadic ductal carcinoma in situ with 18 polymorphic markers from 8p. The min- imal region of loss encompassed the interval be- tween D8S520 and D8S261, and one tumor had loss of D8S550 only. We chose to begin physical mapping of this minimal LOH region by concentrating on the distal end, which includes D8S550. A fine-structure radiation hybrid map for the region that extends from D8S520 (distal) to D8S1759 (proximal) was pre- pared, followed by construction of a single, inte- grated YAC/BAC contig for the interval. The 1730-kb contig consists of 13 YACs and 27 BACs. Fifty-four sequence-tagged sites (STSs) developed from BAC insert end-sequences and 11 expressed sequence tags were localized within the contig by STS content mapping. In addition, four unique cDNA clones from the region were isolated and fully se- quenced. This integrated YAC/BAC resource pro- vides the starting point for transcription mapping, genomic sequencing, and positional cloning of this region. © 1999 Academic Press INTRODUCTION Chromosome loss reveals constitutional recessive mutations that can result in tumor development (Sager, 1989; Marshall, 1991). Individuals heterozy- gous for a mutated tumor suppressor allele express the mutant phenotype (cancer) when the normal allele is lost. Thus, loss of heterozygosity (LOH) analysis may be used to map putative tumor suppressor genes when the DNA from normal and tumor-derived cells for the same patient are compared, using polymorphic mark- ers flanking the locus. Frequent LOH of chromosome 8p has been reported in several types of human cancer, e.g., tumors of the prostate (Vocke et al., 1996), lung (Emi et al., 1992; Wood et al., 1994), colon (Emi et al., 1992; Wood et al., 1994), larynx (Sunwoo et al., 1996), liver (Emi et al., 1992), and breast (Chuaqui et al., 1995; Kerangueven et al., 1995; Adelaide et al., 1998; Dahiya et al., 1998). Studies in colorectal cancer have indicated the pres- ence of at least two tumor suppressor genes (TSG) on 8p, at 8p11–p21.3 and 8p21–p22 (Farrington et al., 1996). Similarly, three distinct regions, at 8p23, 8p22, and 8p12–p21, were defined by LOH studies for pros- tate cancer (Bova et al., 1996; Vocke et al., 1996; Per- inchery et al., 1999) and tumors of the head and neck (Sunwoo et al., 1996; Wu et al., 1997). Wright et al. (1998) identified three regions of loss on 8p (one in 8p22 and two in 8p23) for ovarian adenocarcinomas. Markers in the region at 8p12–p22 have been shown by linkage analysis to have significant lod scores in several German breast cancer families, suggesting that BRCA3 may reside within that region (Seitz et al., Sequence data from this article have been deposited with the EMBL/GenBank Data Libraries under Accession Nos. AF122926 – AF122948, AF124266 –AF124297, and AF124365–AF124368. 1 Present address: Department of Psychiatry, Washington Univer- sity School of Medicine, St. Louis, MO 63110. 2 Present address: Parkcrest Surgical Associates Inc., St. Louis, MO 63141. 3 Present address: Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110. 4 To whom correspondence should be addressed at Department of Psychiatry, Box 8134, Washington University School of Medicine, St. Louis, MO 63110. Telephone: (314) 362-8691. Fax: (314) 747-2983. E-mail: goatea@psychiatry.wustl.edu. 5 Present address: Division of Biology and Biomedical Sciences, Washington University School of Medicine, St. Louis, MO 63110. Genomics 60, 1–11 (1999) Article ID geno.1999.5905, available online at http://www.idealibrary.com on 1 0888-7543/99 $30.00 Copyright © 1999 by Academic Press All rights of reproduction in any form reserved.