Isolation and regional localization of a large collection (2,000) of single-copy DNA fragments on human chromosome 3 for mapping and cloning tumor suppressor genes

M. I. Lerman, F. Latif, G. M. Glenn, L. N. Daniel, H. Brauch, S. Hosoe, K. Hampsch, J. Delisio, M. L. Orcutt, O. W. McBride, K. H. Grzeschik, T. Takahashi, J. Minna, P. Anglard, W. M. Linehan, B. Zbar

Research output: Contribution to journalArticle

60 Scopus citations

Abstract

A collection of 2,000 lambda phage-carrying human single-copy inserts (> 700 bp) were isolated from two chromosome-3 flow-sorted libraries. The single-copy DNA fragments were first sorted into 3p and 3q locations and about 700 3p fragments were regionally mapped using a deletion mapping panel comprised of two humanhamster and two-human-mouse cell hybrids, each containing a chromosome 3 with different deletions in the short arm. The hybrids were extensively mapped with a set of standard 3p markers physically localized or ordered by linkage. The deletion mapping panel divided the short arm into five distinct subregions (A-E). The 3p fragments were distributed on 3p regions as follows: region A, 26%; B, 31%; C, 4%; D, 4% and E, 35%. We screened 300 single-copy DNA fragments from the distal part of 3p (regions A and B) with ten restriction endonucleases for their ability to detect restriction fragment length polymorphisms (RFLPs). Of these fragments 110 (36%) were found to detect useful RFLPs: 35% detected polymorphisms with frequency of heterozygosity of 40% or higher, and 25% with frequency of 30% or higher. All polymorphisms originated from single loci and most of them were of the base pair substitution type. These RFLP markers make it possible to construct a fine linkage map that will span the distal part of chromosome 3p and encompasses the von Hippel-Lindau disease locus. The large number of single-copy fragments (2,000) spaced every 100–150 kb on chromosome 3 will make a significant contribution to mapping and sequencing the entire chromosome 3. The 300 conserved chromosome 3 probes will increase the existing knowledge of man-mouse homologies.

Original languageEnglish (US)
Pages (from-to)567-577
Number of pages11
JournalHuman genetics
Volume86
Issue number6
DOIs
StatePublished - Apr 1991

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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    Lerman, M. I., Latif, F., Glenn, G. M., Daniel, L. N., Brauch, H., Hosoe, S., Hampsch, K., Delisio, J., Orcutt, M. L., McBride, O. W., Grzeschik, K. H., Takahashi, T., Minna, J., Anglard, P., Linehan, W. M., & Zbar, B. (1991). Isolation and regional localization of a large collection (2,000) of single-copy DNA fragments on human chromosome 3 for mapping and cloning tumor suppressor genes. Human genetics, 86(6), 567-577. https://doi.org/10.1007/BF00201543