Isolation and field-inversion gel electrophoresis analysis of DNA markers located close to the Huntington disease gene.
A radiation-induced hybrid cell line containing 10-20 million base pairs of DNA derived from the terminal part of human 4p16 in a background of hamster chromosomes has been used to construct a genomic library highly enriched for human sequences located close to the Huntington disease (HD) gene. Recombinant phage containing human inserts were isolated from this library and used as hybridization probes against two other radiation hybrids containing human fragments with chromosomal breaks in 4p16 and against a human-hamster somatic cell hybrid that retains only the 4p15-4pter part of chromosome 4. Of 121 human phage tested, 6 were mapped distal to the HD-linked D4S10 locus. Since the HD gene is located between D4S10 and the 4p telomere, all of these sequences are likely to be closer to HD than D4S10, and any one of them may be a distal flanking marker for the disease locus. Long-range restriction map analysis performed with a field-inversion gel system shows that the six new loci are distributed in different places within 4p16. Although it is not possible to establish an order for the six sequences with the FIGE data, the results demonstrate that the region detected by these probes must span at least 2000 kb of DNA.
Pritchard
C A
CA
Department of Physiology, University of California, San Francisco 94143.
Casher
D
D
Uglum
E
E
Cox
D R
DR
Myers
R M
RM
D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D002894:Chromosomes, Human, Pair 4 / Q000648:ultrastructure*; D006224:Cricetinae; D003412:Cricetulus; D015342:DNA Probes; D004586:Electrophoresis / Q000379:methods*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005819:Genetic Markers; D006801:Humans; D006816:Huntington Disease / Q000235:genetics*; D006822:Hybrid Cells
D015342:DNA Probes; D005819:Genetic Markers