Segregation of the Huntington disease region of human chromosome 4 in a somatic cell hybrid.
We have developed an X-irradiation:cell fusion procedure that segregates segments of human chromosomes lacking selectable markers and have used this approach to construct somatic cell hybrids retaining fragments of human chromosome 4 as the only human material. To identify hybrids retaining a small chromosomal fragment in the region of the Huntington disease (HD) gene, we used Southern blot analysis to screen 72 hybrid lines for the presence or absence of seven chromosome 4 single-copy loci. These data, combined with in situ hybridization experiments, identified three hybrids of interest. One of these cell lines, C25, stably retains a 10,000- to 20,000-kb fragment of distal 4p in the vicinity of the HD gene, translocated to a hamster chromosome. Field-inversion gel electrophoresis revealed no evidence of rearrangements in the human DNA present in C25. In combination with similar radiation hybrids, C25 is a valuable tool for isolating DNA probes near the HD gene.
Cox
D R
DR
Department of Psychiatry, University of California, San Francisco 94143.
Pritchard
C A
CA
Uglum
E
E
Casher
D
D
Kobori
J
J
Myers
R M
RM
D000818:Animals; D015139:Blotting, Southern; D002459:Cell Fusion; D002874:Chromosome Mapping / Q000379:methods*; D002894:Chromosomes, Human, Pair 4 / Q000648:ultrastructure*; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D006816:Huntington Disease / Q000235:genetics*; D006822:Hybrid Cells