核小体的定点体外组装

Site directed in-vitro assembly of nucleosomes.

作者信息

Wittig S, Wittig B

出版信息

Nucleic Acids Res. 1982 Jun 25;10(12):3647-65. doi: 10.1093/nar/10.12.3647.

DOI:10.1093/nar/10.12.3647

PMID:6287417

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC320741/

Abstract

An in-vitro system is described allowing for the assembly of nucleosomes on preselected sites of a cloned tRNA gene. The system consists of a soluble nucleoprotein fraction, a histone source, and circular DNA containing a single stranded stretch (sssDNA). Nucleosomes assemble on the sssDNA, if the three components are incubated as a highly concentrated solution in the presence of the four deoxyribonucleotidetriphosphates and of ATP. The single stranded stretch is rendered double stranded during incubation. The middle axis of one assembled nucleosome always coincides roughly with the midpoint of the original single stranded DNA stretch.

摘要

本文描述了一种体外系统，该系统能够在克隆的tRNA基因的预选位点上组装核小体。该系统由可溶性核蛋白组分、组蛋白来源以及含有单链延伸段（sssDNA）的环状DNA组成。如果这三种组分在四种脱氧核糖核苷三磷酸和ATP存在的情况下作为高浓度溶液孵育，核小体就会在sssDNA上组装。在孵育过程中，单链延伸段会变成双链。一个组装好的核小体的中轴线总是大致与原始单链DNA延伸段的中点重合。