Optical mapping of protein-DNA complexes on chromatin fibers.

Optical mapping of protein-DNA complexes on chromatin fibers.
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染色质纤维上蛋白质-DNA 复合物的光学图谱。

DOI:
10.1007/978-1-60761-789-1_7
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发表时间:
2010
影响因子:
--
通讯作者:
B. Sullivan
B. Sullivan
中科院分区:
--
文献类型:
--
作者:
B. Sullivan

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免疫荧光(IF)和荧光原位杂交(FISH)是研究中期染色体和间期核结构和组织的常规方法。使用这些技术,整个染色体区域、染色质亚区和特定DNA序列的位置可以在千碱基或兆基的分辨率下进行评估。在延伸的染色质纤维上使用联合IF-FISH可以实现更高的蛋白质和DNA空间关系的分辨率。这种光学作图方法是一种强大的系统,可以沿着释放的染色质纤维定位分子探针,并显示小(<20kb)或大(20-5000kb)的染色体区域。染色质纤维分析可以填补经典染色体研究和分子分析之间的空白,例如在单个或多个核小体水平上评估染色质组织的染色质免疫沉淀(ChIP)。在本章中,介绍了染色质纤维IF-FISH的概念和技术方面,以及成功应用的例子。
Immunofluorescence (IF) and Fluorescence in situ Hybridization (FISH) are conventional methods used to study the structure and organization of metaphase chromosomes and interphase nuclei. Using these techniques, the locations of whole chromosome territories, chromatin subdomains, and specific DNA sequences can be evaluated at kilobase or megabase resolution. Even higher resolution of the spatial relationships of proteins and DNA can be achieved using combined IF-FISH on extended chromatin fibers. This method of optical mapping is a powerful system for localizing molecular probes along released chromatin fibers and visualizing small (<20 kb) or large (20-5,000 kb) chromosomal domains. Chromatin fiber analysis can fill the gaps in resolution between classical chromosome studies and molecular analyses, such as chromatin immunoprecipitation (ChIP) that evaluates chromatin organization at the level of single or multiple nucleosomes. In this chapter, the conceptual and technical aspects of chromatin fiber IF-FISH are presented, along with examples of successful applications.
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发表时间: 1994-04
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