open access

https://hdl.handle.net/2115/81986

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

英語

wheat germ cell‐free protein synthesis

2021-06

John Wiley & Sons

Febs open bio

11(6)

1552-1564

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.

学術雑誌論文

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Creative Commons Attribution 4.0 International

2211-5463