[Amplification of portions of the genome in mammalian somatic cells resistant to colchicine. VI. Restriction analysis of the amplified DNA sequences].
Fragments specific for the amplified regions in DNA of Djungarian hamster colchicine-resistant cells were studied after restriction endonuclease digestion. We used three different methods of detection of these fragments: a) comparison of the wild type and resistant cell DNA electroforegramms stained by ethidium bromide; b) blotting of DNA from sensitive and resistant variants onto nitrocellulose filters and their hybridization with nick-translated DNA from resistant cells, in the presence of the excess of unlabelled DNA from the wild type cells (competitive hybridization); c) investigation of autonomously replicating DNA from sensitive and colchicine-resistant sublines. The highest resolution was found using the third method. However, the competitive hybridization is evidently a more universal approach to restriction analysis of DNA amplified sequences, because it gives quite high resolution and may be used for studying both autonomously and non-autonomously replicating sequences.
D000818:Animals; D002478:Cells, Cultured; D002875:Chromosomes / Q000187:drug effects; D003078:Colchicine / Q000037:antagonists & inhibitors*; D006224:Cricetinae; D004247:DNA / Q000032:analysis / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000494:pharmacology*; D004351:Drug Resistance; D005784:Gene Amplification / Q000187:drug effects*; D005821:Genetic Techniques; D009693:Nucleic Acid Hybridization / Q000187:drug effects; D012091:Repetitive Sequences, Nucleic Acid / Q000187:drug effects*
D004247:DNA; D004262:DNA Restriction Enzymes; D003078:Colchicine
Amplifikatsiia uchastkov gemoma v somaticheskikh kletkakh mlekopitaiushchikh, ustoĭchivykh k kolkhitsinu. Soobshchenie VI. Restriktsionnyĭ analiz amplifitsirovannykh posledovatel'nosteĭ DNK.