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由串联重复克隆DNA片段和核心组蛋白重构的染色质:用于研究高阶结构的模型系统。

Chromatin reconstituted from tandemly repeated cloned DNA fragments and core histones: a model system for study of higher order structure.

作者信息

Simpson R T, Thoma F, Brubaker J M

出版信息

Cell. 1985 Oct;42(3):799-808. doi: 10.1016/0092-8674(85)90276-4.

Abstract

We describe a model system for study of chromatin structure at levels above that of the nucleosome. A series of fragments with lengths ranging from 172 to 207 bp tandemly repeated three to greater than 50 times was prepared; each repeat contains the region important in forming a positioned core particle on a sea urchin 5S rRNA gene upon in vitro association with histones. The tandemly repeated sequences can be studied as linear DNA fragments or as relaxed or supercoiled circular molecules. A number of criteria indicate that nucleosomes position correctly on all the tandemly repeated elements. Measurement of the change in linking number per core particle led to a value of -1.0. Both length and repeat number dependent changes in conformation of the nucleoproteins are observed. We discuss the possibility that some ordered higher level chromatin structure can form with DNA and core histones alone.

摘要

我们描述了一个用于研究高于核小体水平的染色质结构的模型系统。制备了一系列长度在172至207 bp之间的片段,这些片段串联重复三到五十多次;每个重复片段都包含在体外与组蛋白结合时,在海胆5S rRNA基因上形成定位核心颗粒的重要区域。串联重复序列可以作为线性DNA片段或松弛或超螺旋环状分子进行研究。许多标准表明,核小体在所有串联重复元件上都能正确定位。每个核心颗粒连接数变化的测量得出的值为-1.0。观察到核蛋白构象的长度和重复数依赖性变化。我们讨论了仅由DNA和核心组蛋白就能形成一些有序的高级染色质结构的可能性。

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