在哺乳动物DNA合成过程中，复制中间体连接后紧接着存在一个独特的阶段。
There exists a distinct stage during mammalian DNA synthesis immediately after joining of replication intermediates.
作者信息
Lönn U, Lönn S
出版信息
Nucleic Acids Res. 1986 May 12;14(9):3883-94. doi: 10.1093/nar/14.9.3883.
We describe an approach, using alkaline cell lysis and digestion with nuclease S1, which permits to distinguish between newly ligated DNA and the DNA of mature chromatin. When cells with steady-state labelled DNA (mature DNA) are analyzed, the results show labelled "nucleosomal-sized" DNA. However, when DNA of cells pulse-labelled with thymidine for 45 seconds is examined one can detect only large DNA. The newly ligated DNA is not reduced to "nucleosomal-sized" DNA by nuclease S1. When the large DNA is denatured in formamide one can detect 10 kb DNA fragments. Furthermore in pulse-chase experiments there appear, after formamide-treatment, increasing amounts of "nucleosomal-sized" DNA with a parallel decrease in the amount of 10 kb DNA fragments. Hence the newly ligated, large, DNA differs from mature DNA and represents a distinct stage during DNA replication.
我们描述了一种方法，即使用碱性细胞裂解和核酸酶S1消化，该方法能够区分新连接的DNA和成熟染色质的DNA。当分析具有稳态标记DNA（成熟DNA）的细胞时，结果显示出标记的“核小体大小”的DNA。然而，当检查用胸苷脉冲标记45秒的细胞的DNA时，只能检测到大型DNA。新连接的DNA不会被核酸酶S1降解为“核小体大小”的DNA。当大型DNA在甲酰胺中变性时，可以检测到10 kb的DNA片段。此外，在脉冲追踪实验中，经过甲酰胺处理后，“核小体大小”的DNA量增加，同时10 kb DNA片段的量平行减少。因此，新连接的大型DNA与成熟DNA不同，代表了DNA复制过程中的一个独特阶段。
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