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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Agriculture / Faculty of Agriculture >
Peer-reviewed Journal Articles, etc >
De novo reconstitution of chromatin using wheat germ cell-free protein synthesis
This item is licensed under:Creative Commons Attribution 4.0 International
| 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. |
| Rights: | http://creativecommons.org/licenses/by/4.0/ |
| Type: | article |
| URI: | http://hdl.handle.net/2115/81986 |
| 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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