Microdissection and cloning of mouse chromosome 16C3-C4 yields multiple polymorphic markers
- Indiana Univ. School of Medicine, Indianapolis, IN (United States); and others
A regional DNA library of mouse chromosome 16C3-C4 was constructed by microdissection and cloning. This region contains the mouse weaver gene (wv), and is homologous to human chromosomal region (21q21-22). The library was proved to be from the dissected region by fluorescent in situ hybridization (FISH). Based on 50 randomly-picked clones, insert sizes range from 80 to 1280 bp with an average size of over 500 bp. Colony hybridization of 2,000 clones with mouse genomic DNA showed that 59% represented low copy-number sequences. One-hundred such clones were picked randomly for Southern blot analysis with genomic DNA from human, mouse, hamster and a mouse/hamster hybrid containing mouse chromosome 16. Of these, 33 and 17 clones showed a single or multiple bands, respectively, on mouse chromosome 16, 19 were mouse repetitive sequences, and the remaining 31 failed to yield bands. Twenty-eight mouse chromosome 16 single-copy clones were sequenced and tested by Southern blotting for polymorphism with six restriction enzymes (BamHI, EcoRI, HindIII, MspI, PstI, TaqI and XbaI) in mouse strains Balb/C, CBA and C57BI/6J. Fifteen clones showed at least one enzyme polymorphism. Another nine clones, not polymorphic after enzyme digestion, were polymorphic by single stranded conformational polymorphism analysis. Thus 85% (24/28) of the unique chromosome 16 microdissected clones will be useful as polymorphic markers in this 10 to 15 cM region. Approximately one-third of the clones hybrized to human genomic DNA (discrete bands or smears) under conditions of moderate to high stringency.
- OSTI ID:
- 134137
- Report Number(s):
- CONF-941009--
- Journal Information:
- American Journal of Human Genetics, Journal Name: American Journal of Human Genetics Journal Issue: Suppl.3 Vol. 55; ISSN AJHGAG; ISSN 0002-9297
- Country of Publication:
- United States
- Language:
- English
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