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

Kei Fujiwara, Tsutomu Katayama, Shin Ichiro M Nomura

Research output: Contribution to journalArticle

26 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)7176-7183
Number of pages8
JournalNucleic Acids Research
Volume41
Issue number14
DOIs
Publication statusPublished - 2013 Aug
Externally publishedYes

Fingerprint

Bacterial Chromosomes
DNA Replication
DNA Polymerase III
Proteins
DNA Primase
DNA Helicases
RNA Polymerase III
Temperature
DNA
Genes

ASJC Scopus subject areas

  • Genetics

Cite this

Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system. / Fujiwara, Kei; Katayama, Tsutomu; Nomura, Shin Ichiro M.

In: Nucleic Acids Research, Vol. 41, No. 14, 08.2013, p. 7176-7183.

Research output: Contribution to journalArticle

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