Single molecule analysis of DNA replication
Single molecule analysis of DNA replication
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DOI：
10.1016/s0300-9084(00)87170-9
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发表时间：
1999-08-01
期刊：
影响因子：
3.9
通讯作者：
Bensimon, A
中科院分区：
文献类型：
--
作者：
Herrick, J;Bensimon, A
我们在这里描述了一种新的方法来研究DNA复制。该方法基于一种称为分子梳理的过程，并允许对基因组DNA样品中复制单位和复制起点的空间和时间组织进行全基因组分析。分子梳理是一个过程，其中DNA分子通过后退的空气/水界面施加的力在玻璃表面上拉伸和排列。由于拉伸发生在弯月面附近，所有分子以尺寸和顺序无关的方式相同地拉伸。应用荧光杂交梳DNA的结果在一个高分辨率（1至4 kb）的光学映射，是简单的，可控的和可重复的。在单个盖玻片上梳理多达几百个单倍体基因组的能力允许进行统计学上显著数量的测量。直接标记复制的DNA序列反过来使DNA复制的起点可视化和映射。因此，这些功能使分子梳理DNA复制的基因组研究的一个有吸引力的工具。在下文中，我们将讨论分子梳理在DNA复制和基因组稳定性研究中的应用。(C)1999年法国生物化学和生物分子学会/科学和医学版Elsevier SAS。
We describe here a novel approach for the study of DNA replication. The approach is based on a process called molecular combing and allows for the genome wide analysis of the spatial and temporal organization of replication units and replication origins in a sample of genomic DNA. Molecular combing is a process whereby molecules of DNA are stretched and aligned on a glass surface by the force exerted by a receding air/water interface. Since the stretching occurs in the immediate vicinity of the meniscus, all molecules are identically stretched in a size and sequence independent manner. The application of fluorescence hybridization to combed DNA results in a high resolution (1 to 4 kb) optical mapping that is simple, controlled and reproducible. The ability to comb up to several hundred haploid genomes on a single coverslip allows for a statistically significant number of measurements to be made. Direct labeling of replicating DNA sequences in turn enables origins of DNA replication to be visualized and mapped. These features therefore make molecular combing an attractive tool for genomic studies of DNA replication. In the following, we discuss the application of molecular combing to the study of DNA replication and genome stability. (C) 1999 Societe francaise de biochimie et biologie moleculaire/Editions scientifiques et medicales Elsevier SAS.