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An Unnatural Amino Acid-Regulated Growth Controller Based on Informational Disturbance
Version 1
: Received: 8 July 2021 / Approved: 9 July 2021 / Online: 9 July 2021 (09:42:07 CEST)
A peer-reviewed article of this Preprint also exists.
Kato, Y. An Unnatural Amino Acid-Regulated Growth Controller Based on Informational Disturbance. Life 2021, 11, 920. Kato, Y. An Unnatural Amino Acid-Regulated Growth Controller Based on Informational Disturbance. Life 2021, 11, 920.
Abstract
We designed a novel growth controller regulated by feeding of an unnatural amino acid, Nε-benzyloxycarbonyl-L-lysine (ZK), using a specific incorporation system at a sense codon. This system is constructed by a pair of modified pyrrolisyl-tRNA synthetase (PylRS) and its cognate tRNA (tRNApyl). Although ZK is non-toxic for normal organisms, the growth of Escherichia coli carrying the ZK incorporation system was inhibited in a ZK concentration-dependent manner without causing rapid bacterial death, presumably due to generation of non-functional or toxic proteins. The extent of growth inhibition strongly depended on the anticodon sequence of the tRNApyl gene. Taking advantage of the low selectivity of PylRS for tRNApyl anticodons, we experimentally determined the most effective anticodon sequence among all 64 nucleotide sequences in the anticodon region of tRNApyl gene. The results suggest that the ZK-regulated growth controller is a simple, target-specific, environmental noise-resistant and titratable system. This technique may be applicable to a wide variety of organisms because the growth inhibitory effects are caused by “informational disturbance”, in which the highly conserved system for transmission of information from DNA to proteins is perturbed.
Keywords
synthetic biology; genetic parts; kill-switch; growth control; unnatural amino acids; codon reassignment; pyrrolysine
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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