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

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 of "Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system" by Fujiwara K, Katayama T, Nomura SM, https://doi.org/10.1093/nar/gkt489, used under its Creative Commons Attribution license (CC-BY); markup removed, otherwise unmodified.