Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system.
Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system.
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DOI：
10.1093/nar/gkt489
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发表时间：
2013-08
影响因子：
14.9
通讯作者：
Nomura SM
中科院分区：
文献类型：
--
作者：
Fujiwara K;Katayama T;Nomura SM
所有活细胞的复制都依赖于中心法则的多轮流动。特别是，DNA复制蛋白的表达是中心法则循环过程的关键步骤。在这里，我们通过重组转录-翻译系统（PURE系统）的染色体DNA复制机制的自主表达实现了整个顺序转录-翻译-复制过程。我们发现低温对于使用PURE系统在单管中表达复杂蛋白DNA聚合酶III是必不可少的。将编码启动器、DNA解旋酶、解旋酶装载器、RNA引物酶和DNA聚合酶III的13个基因加入到PURE系统中，通过耦合方式形成DNA复制系统。人工遗传电路表明，由于复制而产生的DNA能够为蛋白质提供遗传信息，表明体外中心法则可以依次经历两轮。
Replication of all living cells relies on the multirounds flow of the central dogma. Especially, expression of DNA replication proteins is a key step to circulate the processes of the central dogma. Here we achieved the entire sequential transcription–translation–replication process by autonomous expression of chromosomal DNA replication machineries from a reconstituted transcription–translation system (PURE system). We found that low temperature is essential to express a complex protein, DNA polymerase III, in a single tube using the PURE system. Addition of the 13 genes, encoding initiator, DNA helicase, helicase loader, RNA primase and DNA polymerase III to the PURE system gave rise to a DNA replication system by a coupling manner. An artificial genetic circuit demonstrated that the DNA produced as a result of the replication is able to provide genetic information for proteins, indicating the in vitro central dogma can sequentially undergo two rounds.
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影响因子：
4.8
作者：
Abe, Yoshito;Jo, Takaaki;Ueda, Tadashi
通讯作者：
Ueda, Tadashi
影响因子：
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作者：
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通讯作者：
Forster, Anthony C.
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作者：
Fujiwara K;Nomura SM
通讯作者：
Nomura SM
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通讯作者：
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