Transfer RNA synthesis-coupled translation and DNA replication in a reconstituted transcription/translation system

Ryota Miyachi; Yoshihiro Shimizu; Norikazu Ichihashi

doi:10.51094/jxiv.35

##article.authors##

Ryota Miyachi Graduate School of Arts and Science, The University of Tokyo
Yoshihiro Shimizu RIKEN Center for Biosystems Dynamics Research
Norikazu Ichihashi Graduate School of Arts and Science, The University of Tokyo

DOI:

https://doi.org/10.51094/jxiv.35

キーワード:

tRNA、 translation、 DNA replication、 cell-free synthetic biology、 PURE system

抄録

Transfer RNAs (tRNA) are key molecules involved in translation. In vitro synthesis of tRNAs and their coupled translation are important challenges in the construction of a self-regenerative molecular system. Here, we first purified EF-Tu and ribosome components in a reconstituted translation system of Escherichia coli to remove residual tRNAs. Next, we expressed 15 types of tRNAs in the repurified translation system and performed translation of the reporter luciferase gene depending on the expression. Furthermore, we demonstrated DNA replication through expression of a tRNA encoded by DNA, mimicking information processing within the cell. Our findings highlight the feasibility of an in vitro self-reproductive system in which tRNAs can be synthesized from replicating DNA.

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