DELINEATION OF DNA-REPLICATION TIME ZONES BY FLUORESCENCE INSITU HYBRIDIZATION
DELINEATION OF DNA-REPLICATION TIME ZONES BY FLUORESCENCE INSITU HYBRIDIZATION
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DOI：
10.1002/j.1460-2075.1992.tb05162.x
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发表时间：
1992-03-01
期刊：
影响因子：
11.4
通讯作者：
CEDAR, H
中科院分区：
文献类型：
--
作者：
SELIG, S;OKUMURA, K;CEDAR, H
荧光原位杂交已被用于可视化间期核中的特定基因组DNA序列。在正常的二倍体细胞中，未复制的DNA片段产生单态杂交信号，而复制的基因座以双态为特征。这两种模式在非同步化细胞群体中的分布可用于确定任何DNA序列的S期复制时间。这种方法的有效性是通过分析基因建立的，这些基因在表达和非表达细胞中的复制谱先前已经通过常规方法确定。使用这种技术，它已经有可能映射的复制时间拓扑结构的DNA内和侧翼的囊性纤维化（CF）基因座7号染色体上。该基因本身位于一个确定的时区内，其长度约为500 kb，并处于发育控制之下。它在表达CF的细胞中早期复制，但在其他细胞类型中晚期复制。这些时区可能代表了染色体结构的基本单位。
Fluorescence in situ hybridization has been used to visualize specific genomic DNA sequences in interphase nuclei. In normal diploid cells, unreplicated DNA segments give singlet hybridization signals while replicated loci are characterized by doublets. The distribution of these two patterns in unsynchronized cell populations can be used to determine the S phase replication time of any DNA sequence. The validity of this approach was established by analyzing genes whose replication profiles in expressing and non-expressing cells had been determined previously by conventional methods. Using this technique it has been possible to map the replication timing topography of the DNA within and flanking the cystic fibrosis (CF) gene locus on chromosome 7. The gene itself is located within a defined time zone which is approximately 500 kb in length and is under developmental control. It is early replicating in cells which express CF but late replicating in other cell types. These time zones probably represent basic units of chromosome structure.