| Budget Amount *help |
¥18,720,000 (Direct Cost: ¥14,400,000、Indirect Cost: ¥4,320,000)
Fiscal Year 2019: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2017: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2016: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
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| Outline of Final Research Achievements |
Self-replication is a fundamental feature of living cells. Cells produce their own copy based on their genetic information. We recently developed an in vitro reconstitution system termed RCR (replication cycle reaction), in which replication cycle of the Escherichia coli chromosome repeats continuously to propagate a mini-chromosome that is a circular DNA having replication origin, oriC. RCR consists of 26 proteins .
In this study, we combined an in vitro transcription-translation reconstitution system, PURE system with RCR (PURE-coupled RCR). All 26 RCR proteins were able to express functionally from DNA in PURE system. We constructed a mini-chromosome encoding the RCR proteins. This mini-chromosome allow us to propagate the circular DNA depending on the transcription-translation from its own DNA. We further developed a system in which the PURE-coupled RCR progress from a single mini-chromosome molecule in water-in-oil emulsion for purpose of in vitro evolution of the mini-chromosome.
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