Gudkov A V, Kopnin B P
Genetika. 1983 Jul;19(7):1045-53.
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.
在用限制性内切酶消化后，对中仓鼠秋水仙碱抗性细胞DNA中扩增区域的特异性片段进行了研究。我们使用了三种不同的方法来检测这些片段：a）比较经溴化乙锭染色的野生型和抗性细胞DNA电泳图谱；b）将敏感型和抗性变体的DNA印迹到硝酸纤维素滤膜上，并在存在过量野生型细胞未标记DNA的情况下，使其与来自抗性细胞的缺口平移DNA杂交（竞争性杂交）；c）研究敏感型和秋水仙碱抗性亚系的自主复制DNA。发现使用第三种方法具有最高的分辨率。然而，竞争性杂交显然是对DNA扩增序列进行限制性分析的一种更通用的方法，因为它具有相当高的分辨率，并且可用于研究自主和非自主复制序列。
