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De novo reconstitution of chromatin using wheat germ cell-free protein synthesis

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Title: De novo reconstitution of chromatin using wheat germ cell-free protein synthesis
Authors: Endo, Yaeta Browse this author
Takemori, Nobuaki Browse this author
Nagy, Szilvia K. Browse this author
Okimune, Kei-ichi Browse this author
Kamakaka, Rohinton Browse this author
Onouchi, Hitoshi Browse this author
Takasuka, Taichi E. Browse this author →KAKEN DB
Keywords: ATP‐dependent nucleosome assembly
histone modifications
Mg2+ chromatin isolation
post‐translational chromatin assembly
unmodified and soluble histone products
wheat germ cell‐free protein synthesis
Issue Date: Jun-2021
Publisher: John Wiley & Sons
Journal Title: Febs open bio
Volume: 11
Issue: 6
Start Page: 1552
End Page: 1564
Publisher DOI: 10.1002/2211-5463.13178
Abstract: DNA is packaged with histones to form chromatin that impinges on all nuclear processes, including transcription, replication and repair, in the eukaryotic nucleus. A complete understanding of these molecular processes requires analysis of chromatin context in vitro. Here, Drosophila four core histones were produced in a native and unmodified form using wheat germ cell-free protein synthesis. In the assembly reaction, four unpurified core histones and three chromatin assembly factors (dNAP-1, dAcf1 and dISWI) were incubated with template DNA. We then assessed stoichiometry with the histones, nucleosome arrays, supercoiling and the ability of the chromatin to serve as a substrate for histone-modifying enzymes. Overall, our method provides a new avenue to produce chromatin that can be useful in a wide range of chromatin research.
Type: article
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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