Eukaryotic DNA replication with purified budding yeast proteins

被引:6
|
作者
Posse, Viktor [1 ,2 ]
Johansson, Erik [3 ]
Diffley, John F. X. [1 ]
机构
[1] Francis Crick Inst, Chromosome Replicat Lab, London, England
[2] Univ Gothenburg, Dept Med Biochem & Cell Biol, Gothenburg, Sweden
[3] Umea Univ, Dept Med Biochem & Biophys, Umea, Sweden
来源
基金
英国惠康基金; 英国医学研究理事会; 瑞典研究理事会; 欧洲研究理事会;
关键词
REPLISOME;
D O I
10.1016/bs.mie.2021.08.018
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The in vitro reconstitution of origin firing was a key step toward the biochemical reconstitution of eukaryotic DNA replication in budding yeast. Today the basic replication assay involves proteins purified from 24 separate protocols that have evolved since their first publication, and as a result, the efficiency and reliability of the in vitro replication system has improved. Here we will present protocols for all 24 purifications together with a general protocol for the in vitro replication assay and some tips for troubleshooting problems with the assay.
引用
收藏
页码:1 / 33
页数:33
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