2016 Fiscal Year Final Research Report
Propagation of large circular DNA by reconstitution of a chromosome-replication cycle
| Project/Area Number |
26640116
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Genome biology
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| Research Institution | Rikkyo University |
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Principal Investigator |
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Keywords | DNA複製 / ゲノム / 大腸菌染色体 / 合成生物学 / 合成ゲノム学 |
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| Outline of Final Research Achievements |
Escherichia coli has a 4.6 Mb circular chromosome with a replication origin, oriC. While the oriC replication has been reconstituted in vitro more than 30 years ago, continuous repetition of the replication cycle has not yet been achieved. Here, we reconstituted the entire replication cycle with purified enzymes that catalyze initiation at oriC, bidirectional fork progression, Okazaki-fragment maturation, and decatenation of the replicated circular products. Because decatenation provides covalently closed supercoiled monomers that are competent for the next round of replication initiation, the replication cycle repeats autonomously and continuously in an isothermal condition. This Replication-Cycle Reaction (RCR) propagates large circular DNA up to 0.2 Mb as intact covalently closed molecules. RCR provides a powerful in vitro tool to propagate large circular DNA molecules
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| Free Research Field |
大腸菌ゲノム複製
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