Detection of recombinational mutations in cultured human cells by Southern blot analysis with minisatellite DNA probes.
Using the human acute monocytic leukemia cell line, THP-1, a hypermutability of minisatellite loci was demonstrated in cell culture by Southern blot analysis with minisatellite DNA probes. DNA was isolated from 98 subclones and hybridized to a panel of minisatellite probes consisting of three multilocus minisatellite probes (ML probes) and seven locus-specific minisatellite probes (LS probes). The Southern blot patterns of the hybridized subclones were compared with those of the parental THP-1. Four mutated bands with two ML probes and two mutated bands with two LS probes were detected. The mutation frequency was estimated roughly at 0.1% based on the total number of bands analyzed, and it was much higher than that expected for other DNA regions. Four of these mutations were thought to be alterations of repetitions caused by insertion or deletion of tandem repeats, and one mutant lost a complete minisatellite allele. The nature of the sixth mutant was unclear. Because of the hypermutability of minisatellite DNA, Southern blot analysis using minisatellite DNA probes can be used as a mutation assay system directly based on the DNA.
Honma
M
M
Division of Genetics and Mutagenesis, National Institute of Hygienic Sciences, Tokyo, Japan.
Kataoka
E
E
Ohnishi
K
K
Kikuno
T
T
Hayashi
M
M
Sofuni
T
T
Mizusawa
H
H
D001483:Base Sequence; D015139:Blotting, Southern; D016172:DNA Fingerprinting; D015342:DNA Probes; D004273:DNA, Neoplasm; D004276:DNA, Satellite / Q000235:genetics*; D006801:Humans; D007948:Leukemia, Monocytic, Acute; D008297:Male; D008969:Molecular Sequence Data; D009152:Mutagenicity Tests; D009154:Mutation; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D004273:DNA, Neoplasm; D004276:DNA, Satellite