In vitro reconstitution and analysis of mononucleosomes containing defined DNAs and proteins
In vitro reconstitution and analysis of mononucleosomes containing defined DNAs and proteins
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DOI：
10.1006/meth.1997.0441
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发表时间：
1997-05-01
期刊：
影响因子：
4.8
通讯作者：
Lee, KM
中科院分区：
文献类型：
--
作者：
Hayes, JJ;Lee, KM
越来越多地，对真核细胞核内发生的过程的生化分析，如转录和复制，需要构建特定的染色质底物。在体外重组的核小体复合体是最近关于聚合酶进展和反式作用因子结合活性的各种研究的关键元件。重组的复合体可以很容易地从纯化的组分中构建出来，其数量适合于生化和生物物理研究。此外，重组的单核小体复合体表现出天然的生化和生物物理性质，但在蛋白质和DNA组分方面的异质性必然比从自然来源分离的整体复合体要少得多。这篇综述详细介绍了包含独特DNA序列和特定定制的核心组蛋白的模型单核体复合体的重建方案，并描述了与这些程序相关的常见陷阱。(C)1997年学术出版社。
Increasingly, biochemical analyses of processes that occur within eukaryotic nuclei such as transcription and replication require the construction of specific chromatin substrates. Nucleosome complexes reconstituted in vitro have been key elements in a variety of recent studies of polymerase progression and trans-acting factor binding activities. Reconstituted complexes can be easily constructed from purified components in quantities suitable for biochemical and biophysical studies. In addition, reconstituted mononucleosome complexes exhibit native biochemical and biophysical properties but necessarily contain much less heterogeneity with regard to both protein and DNA components than bulk complexes isolated from natural sources. This review details the protocols for reconstitution of model mononucleosome complexes that contain unique DNA sequences and specifically tailored core histone proteins and describes common pitfalls associated with these procedures. (C) 1997 Academic Press.