In Vitro Chromatin Assembly: Strategies and Quality Control

被引：36

作者：

Muthurajan, U. ^{[1
]}

Mattiroli, F. ^{[1
]}

Bergeron, S. ^{[1
]}

Zhou, K. ^{[1
,5
]}

Gu, Y. ^{[1
,5
]}

Chakravarthy, S. ^{[2
]}

Dyer, P. ^{[1
]}

Irving, T. ^{[2
]}

Luger, K. ^{[1
,3
,4
]}

机构：

[1] Univ Colorado, Boulder, CO 80309 USA

[2] IIT, Chicago, IL 60616 USA

[3] Univ Colorado, Howard Hughes Med Inst, Boulder, CO 80309 USA

[4] Colorado State Univ, Inst Genome Architecture & Funct, Ft Collins, CO 80523 USA

[5] Colorado State Univ, Ft Collins, CO 80523 USA

来源：

ENZYMES OF EPIGENETICS, PT A | 2016年 / 573卷

关键词：

NUCLEOSOME CORE PARTICLE; STRUCTURAL-CHARACTERIZATION; CRYSTAL-STRUCTURES; HISTONE; DNA; PROTEIN; FLUORESCENCE; BINDING; MECHANISMS; STABILIZES;

D O I：

10.1016/bs.mie.2016.01.002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Chromatin accessibility is modulated by structural transitions that provide timely access to the genetic and epigenetic information during many essential nuclear processes. These transitions are orchestrated by regulatory proteins that coordinate intricate structural modifications and signaling pathways. In vitro reconstituted chromatin samples from defined components are instrumental in defining the mechanistic details of such processes. The bottleneck to appropriate in vitro analysis is the production of high quality, and quality-controlled, chromatin substrates. In this chapter, we describe methods for in vitro chromatin reconstitution and quality control. We highlight the strengths and weaknesses of various approaches and emphasize quality control steps that ensure reconstitution of a bona fide homogenous chromatin preparation. This is essential for optimal reproducibility and reliability of ensuing experiments using chromatin substrates.

引用

页码：3 / 41

页数：39

共 56 条

[1] Nucleosomes containing the histone variant H2A.Bbd organize only 118 base pairs of DNA [J].