Generation of a panel of somatic cell hybrids containing unselected fragments of human chromosome 10 by X-ray irradiation and cell fusion: application to isolating the MEN2A region in hybrid cells.
We have used X-ray irradiation and cell fusion to generate somatic cell hybrids containing fragments of human chromosome 10. Our experiments were directed towards isolating the region of the MEN2A gene in hybrids and to use those as the source of DNA for cloning and mapping new markers from near the MEN2A locus. A number of hybrid clones containing human sequences that are tightly linked to the MEN2A gene were identified. Some 25% of our hybrids, however, proved to contain more than one human chromosome 10-derived fragment or showed evidence of deletions and/or rearrangements. A detailed analysis of the human content of X-ray irradiation hybrids is required to assess the integrity and number of human fragments retained. Despite retention of multiple human-derived fragments, these hybrids will prove useful as cloning and mapping resources.
Goodfellow
P J
PJ
Department of Medical Genetics, University of British Columbia, Vancouver, Canada.
Povey
S
S
Nevanlinna
H A
HA
Goodfellow
P N
PN
D000818:Animals; D002459:Cell Fusion / Q000528:radiation effects; D002460:Cell Line; D002874:Chromosome Mapping; D002879:Chromosomes, Human, Pair 10 / Q000032:analysis / Q000528:radiation effects / Q000648:ultrastructure*; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000235:genetics / Q000528:radiation effects / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D005819:Genetic Markers / Q000032:analysis; D006801:Humans; D006822:Hybrid Cells / Q000032:analysis / Q000528:radiation effects / Q000648:ultrastructure*; D009377:Multiple Endocrine Neoplasia / Q000032:analysis / Q000235:genetics / Q000648:ultrastructure*; D009693:Nucleic Acid Hybridization; D012150:Polymorphism, Restriction Fragment Length; D014965:X-Rays; D014998:Y Chromosome / Q000032:analysis / Q000528:radiation effects / Q000648:ultrastructure
D005819:Genetic Markers; D004247:DNA