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Simian virus 40 DNA replication in vitro: purification and characterization of replication factors from mouse cells.

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Title: Simian virus 40 DNA replication in vitro: purification and characterization of replication factors from mouse cells.
Authors: Ariga, Hiroyoshi Browse this author →KAKEN DB
Issue Date: 9-Dec-1986
Publisher: Oxford University Press
Journal Title: Nucleic acids research
Volume: 14
Issue: 23
Start Page: 9457
End Page: 9470
Publisher DOI: 10.1093/nar/14.23.9457
PMID: 3025815
Abstract: We have previously developed simian virus 40 (SV40) DNA replication system in vitro (Ariga and Sugano, J. Virol. 48, 481, 1983). This system is composed of human HeLa or mouse FM3A nuclear extract and cytoplasmic extract of SV40 infected CosI cells. Here FM3A nuclear extract was fractionated by DEAE Sephacel and single-stranded DNA cellulose chromatography into three components required for accurate in vitro SV40 DNA replication. One fraction (A fraction) contained DNA polymerase-primase, and the second component (B fraction) contained DNA topoisomerase. Third component was further purified to near homogenuity using DEAE-Sephacel, single-stranded DNA cellulose, and glycerol gradient centrifugation. The purified protein (named factor I) bound to the origin containing fragment of SV40 DNA. The factor I enhanced the initiation of SV40 DNA replication catalyzed by SV40 infected CosI cytoplasm alone. When all four fractions consisting of A, B fractions, factor I, and SV40 infected CosI cytoplasm were mixed together, the system was reconstituted, meaning that initiation and subsequent elongation were completed to generate the full sized daughter molecules.
Type: article
URI: http://hdl.handle.net/2115/54050
Appears in Collections:薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 有賀 寛芳

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