In vitro transcription/translation-coupled DNA replication through the regeneration of 20 aminoacyl-tRNA synthetases
キーワード:regeneration、 self-reproduction、 in vitro synthetic biology、 aminoacyl-tRNA synthetase、 artificial cell
The in vitro reconstruction of life-like self-reproducing systems is a major challenge in in vitro synthetic biology. Self-reproduction requires regeneration of all molecules involved in DNA replication, transcription, and translation. This study demonstrated the DNA replication and regeneration of major translation factors, 20 aminoacyl-tRNA synthetases (aaRS), in a reconstituted transcription/translation system (PURE system). First, we replicated each DNA that encode one of the 20 aaRSs through aaRS expression from the DNA (i.e., regeneration) by serial transfer experiments. Thereafter, we successively increased the number of aaRS genes and achieved simultaneous DNA replication and regeneration of all 20 aaRSs, which comprised approximately half the number of protein factors in the PURE system, except for ribosomes, by employing dialyzed reaction and sequence optimization. This study provides a step-by-step methodology for increasing the number of self-regenerative genes in self-reproducing artificial systems.
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投稿日時: 2022-04-20 07:03:04 UTC
公開日時: 2022-04-20 10:55:52 UTC
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