# Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system

Article · Fujiwara K, Katayama T, Nomura SM

- Archive id: doi:10.1093/nar/gkt489
- Published: 2013-06-03
- Venue: Nucleic acids research
- DOI: https://doi.org/10.1093/nar/gkt489
- PMID: 23737447
- PMCID: PMC3737561
- Citations: 49
- Source URL: https://doi.org/10.1093/nar/gkt489
- PDF: https://europepmc.org/articles/PMC3737561/pdf
- Provenance: europepmc

## Abstract

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.

_Abstract reproduced under CC-BY (article license as recorded by Europe PMC; abstract reproduced unmodified apart from markup removal)._

Topics: [Synthetic Biology & Genome Engineering](https://space-kitty.com/topics/synthetic-biology)

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