Transfer RNA Synthesis-Coupled Translation and DNA Replication in a Reconstituted Transcription/Translation System

Transfer RNAs (tRNAs) 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.