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Radiation hybrid panel for human chromosome 19

Journal Article · · American Journal of Human Genetics
OSTI ID:134802
; ;  [1]
  1. Rhode Island Hospital, Providence, RI (United States)
Radiation hybrid cell lines have proven to be excellent resources for the physical mapping and the isolation of markers from specific chromosomal regions. We have produced radiation hybrids from a monochromosomal microcell hybrid containing chromosome 19. The human chromosome was marked with a retroviral vector containing a dominant selectable marker. The hybrids were produced using doses of radiation ranging from 1000 to 8000 rads and selection for the exogenous marker. A panel of approximately 90 hybrids was characterized using marker analysis. In addition, a subset was analyzed by fluorescent in situ hybridization (FISH). These methods allowed us to identify the number of chromosome fragments and the region of the chromosome contained in the hybrids. The panel has been tested for 50 chromosome 19 markers from known locations on the chromosome using the polymerase chain reaction and Southern blotting. We will continue to test the hybrid panel for additional markers. Microsatellite markers as well as anonymous DNA sequences and genes have been tested. Hybrids have been isolated which appear to contain only 1 or 2 markers from the short arm of the chromosome where the selectable marker is inserted. Cloning of the DNA flanking the exogenous retrovirus was carried out by taking advantage of the tRNA suppressor gene incorporated into the vector. FISH of the phage clone containing flanking DNA hybridized to metaphase spreads of the hybrid localized the insertion site to 19p13.1. The hybrid panel that we have produced will aid in the physical mapping of chromosome 19 and region-specific cloning.
OSTI ID:
134802
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