[Isolation of DNA probes for the detection of sequences amplified in colchicine-resistant cells].
Earlier we have found that the development of resistance to colchicine in mammalian cells in vitro is due to gene amplification leading to decreased plasma membrane permeability to the selective agent and some other unrelated drugs. By a stepwise self-renaturation procedure followed by chromatography on hydroxyapatite we isolated the fraction of middle-repeated sequences (DNAc0t = 10-250) enriched in amplified DNA from the DNA of colchicine-resistant Djungarian hamster cell line. Blotting-hybridization with [32P]DNAc0t = 10-250 performed in the presence of the excess of unlabelled DNA from wild type cells reveals amplified sequences in resistant cell lines. The comparison of DNAs from cell lines resistant to colchicine, adriablastin and actinomycin D showed that common but not identical DNA sequences are amplified in these cases. In situ hybridization with [3H]DNAc0t = 10-250 indicates that amplified sequences are located in the long homogeneously staining regions (HSRs) of the marker chromosomes. These results suggest that DNAc0t = 10-250 may be used for screening of recombinant molecules containing amplified sequences.
Gudkov
A V
AV
Chernova
O B
OB
Sianova
E Iu
EIu
Sokova
O I
OI
Kopnin
B P
BP
D000818:Animals; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology*; D006224:Cricetinae; D004247:DNA / Q000235:genetics*; D004351:Drug Resistance; D005784:Gene Amplification; D005819:Genetic Markers; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid
D005819:Genetic Markers; D004247:DNA; D003078:Colchicine
Poluchenie DNK-zonda, predstavliaiushchego posledovatel'nosti, amplifitsirovannye v ustoĭchivykh k kolkhitsinu kletkakh.