Pulsed field analysis of human hprt large deletion mutants
Journal Article
·
· Environmental and Molecular Mutagenesis
OSTI ID:91982
- Univ. of Vermont, Burlington, VT (United States)
In order to measure extent of deletion and breakpoint locations in hprt large deletion mutants, the hprt chromosomal region (Xq26) was physical mapped using pulsed field gel electrophoresis (PFGE) with five rare-cutter enzymes and 14 linked markers. New linked markers were derived from yeast artificial chromosomes (YAC`s) with Xq26 inserts. Three new YAC-derived DNA markers (DXS1327, DXS1328, and DXS1329) were obtained by preparing genomic DNA libraries from 2 YAC-containing yeast clones. Libraries were initially screened with a human Alu repetitive element probe to identify recombinants were screened for utility as DNA hybridization probes. Four additional YAC derived probes (LyHPRT, RyHPRT, 837L, and 342L) were obtained from Dr. James Fuscoe as PCR primers. New markers were ordered relative to previously available Xq26 probes by deletion mapping with an array of hprt large deletion mutants. The probe order is cen.-DXS53-DXS79-LyHPRT-{sup 5}{prime}hprt{sup 3}{prime}-DXS1327-RyHPRT-DXS1328-837L-(DXS311,DXS1329)-DXS86-342L-DXS10-DXS177-tel. The order of DXS311 relative to DXS1329 is ambiguous as these probes are very closely linked. Linked markers were used to complete a 1.5 megabase physical map of the region telomeric to hprt. Deletion extent and a breakpoint spectrum were determined with 20 hprt 3{prime} deletions using PFGE. Deletion size in these mutants ranged from 30kb to 1,600kb with smaller deletions being more common than larger deletions. Several mutants are non-linear or complex events. This work begins to describe a high resolution breakpoint spectrum of this large chromosomal region in an attempt to gain insight into mechanisms of deletion in human cells.
- OSTI ID:
- 91982
- Report Number(s):
- CONF-9405324--
- Journal Information:
- Environmental and Molecular Mutagenesis, Journal Name: Environmental and Molecular Mutagenesis Journal Issue: Suppl.23 Vol. 23; ISSN 0893-6692; ISSN EMMUEG
- Country of Publication:
- United States
- Language:
- English
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