Competition of aminoacyl-tRNA synthetases for tRNA ensures the accuracy of aminoacylation. Minimization of Elements for Isothermal DNA Replication by an Evolutionary Approach. A partially self-regenerating synthetic cell. Probing self-regeneration of essential protein factors required for: In vitro translation activity by serial transfer. In vitro synthesis of 32 translation-factor proteins from a single template reveals impaired ribosomal processivity. In vitro self-replication and multicistronic expression of large synthetic genomes. Libicher, K., Hornberger, R., Heymann, M. De novodesign and synthesis of a 30-cistron translation-factor module. Cogenerating Synthetic Parts toward a Self-Replicating System. Activities of 20 aminoacyl-tRNA synthetases expressed in a reconstituted translation system in Escherichia coli. Cell-free translation reconstituted with purified components. Cell-Free Biocatalysis for the Production of Platform Chemicals. Gene Expression Inside Liposomes: From Early Studies to Current Protocols. Connecting primitive phase separation to biotechnology, synthetic biology, and engineering. Reconstituting Natural Cell Elements in Synthetic Cells. Cell-free gene expression: an expanded repertoire of applications. The hallmarks of living systems: Towards creating artificial cells. Artificial Cells: Synthetic Compartments with Life-like Functionality and Adaptivity. What can we learn from the construction of in vitro replication systems? Ann. Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology. Modularize and Unite: Toward Creating a Functional Artificial Cell. Bottom-Up Synthesis of Artificial Cells: Recent Highlights and Future Challenges. Compartmentalizing cell-free systems: Toward creating life-like artificial cells and beyond. On the “Life-Likeness” of Synthetic Cells. This study provides a step-by-step methodology for increasing the number of self-regenerative genes in self-reproducing artificial systems. 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. 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. 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). Self-reproduction requires regeneration of all molecules involved in DNA replication, transcription, and translation. The in vitro reconstruction of life-like self-reproducing systems is a major challenge in in vitro synthetic biology. Regeneration、 self-reproduction、 in vitro synthetic biology、 aminoacyl-tRNA synthetase、 artificial cell 抄録 Graduate School of Arts and Science, The University of Tokyo
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