2021年6月
De novo reconstitution of chromatin using wheat germ cell‐free protein synthesis
FEBS Open Bio
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- 巻
- 11
- 号
- 6
- 開始ページ
- 1552
- 終了ページ
- 1564
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1002/2211-5463.13178
- 出版者・発行元
- Wiley
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.
- リンク情報
- ID情報
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- DOI : 10.1002/2211-5463.13178
- ISSN : 2211-5463
- eISSN : 2211-5463
- PubMed ID : 33960726
- PubMed Central 記事ID : PMC8167859