著者

末次 正幸

出版者

一般社団法人 日本生物物理学会

雑誌

生物物理 (ISSN:05824052)

巻号頁・発行日

vol.60, no.5, pp.284-287, 2020 (Released:2020-09-30)

参考文献数

15

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In vitro reconstitution of cellular events is a promising strategy as a bottom-up approach to build an artificial cell in the field of synthetic biology. Also, the reconstituted system sometimes provides a useful biotechnology tool for various fields. We recently reconstituted a whole replication cycle of the Escherichia coli chromosome in vitro as a part of the bottom-up synthetic biology. This replication cycle reaction (RCR) propagates circular DNA exponentially and precisely by autonomous repetition of the cycle in an isothermal reaction. Even genome-sized DNA (1 Mbp) can be propagated as circular DNA in RCR. We also developed a DNA assembly tool. The combination of the DNA assembly and RCR amplification enables cell-free synthesis of artificial large circular DNA, providing a powerful tool for a top-down study of the synthetic biology, in which living cells are modified at a whole genome level.