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Biological Carcinogenesis and Development Program, Science Applications International Corporation-Frederick [M-H. W., F-M. D., L. G.], and Laboratory of Immunobiology [F. L., I. K., B. Z., M. I. L.], National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, Maryland 21702; Simmons Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas 75235 [S. B. J-Y. C., Y. S., J. D. M.]; Department of Tumor Biology, Karolinska Institute, Stockholm 17177, Sweden [V. K., E. Z., G. K.]; and Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030 [C-C. L.]
The critical region on human chromosome 3p21.3 harboring a putative lung cancer tumor suppressor gene (TSG) was previously defined by allelotyping and recently refined by overlapping homozygous deletions. We report the construction of a 700-kb (cosmid and one P1 phage) clone contig covering the deletion overlap and its flanks. The minimal set of 23 cosmids comprises 600 kb and is extended by one P1 phage to 700 kb to cover the distal breakpoint of the overlap. The clone contig was extensively characterized by restriction and expression mapping to produce high resolution physical and transcription maps of the cloned region. Potential transcribed fragments were detected by hybridization with PCR-amplified cDNA libraries, direct cDNA selection, "zoo" blotting, cDNA screening, and identification of 24 CpG islands. Thus far, 15 new genes represented by partial or full-length cDNAs were isolated, characterized, and precisely positioned on the contig. Two previously cloned genes, namely GNAI-2 and GNAT-1, were also positioned. In addition, the telomeric breakpoint of the NCI H740 deletion and the centromeric breakpoint of the overlapping GLC20 deletion were discovered and mapped to define precisely the candidate TSG region. This large cosmid clone contig and high resolution maps will prove crucial in the identification of the lung cancer TSG(s).
1 The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. government.
2 Supported by the Nancy B. and Jake L. Hamon Center for Therapeutic Oncology Research Foundation, the G. Harold and Leila Y. Mathers Charitable Foundation, the Leroy "Skip" Malouf Foundation, the Julie Gould Foundation, the W. A. "Tex" and Deborah Moncrief, Jr., Center for Cancer Genetics Foundation, and National Cancer Institute Grant P20 CA58220.
3 Supported by the Swedish Cancer Society and by grants from the Cancer Research Institute/Concern Foundation for Cancer Research and Karolinska Institute.
4 To whom requests for reprints may be addressed, at Laboratory of Immunobiology, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, MD 21702.
5 To whom requests for reprints also may be addressed, at Simmons Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX 75235.
Received 10/30/95. Accepted 2/15/96.
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