Activation of a diluted E. coli cell-free transcription-translation system within liposomes by hypertonic concentration

Matsui, Yukino; Akui, Toshiki; Doi, Nobuhide; Fujiwara, Keigi

doi:10.1016/j.xpro.2021.101003

Cited by 1 publication

(1 citation statement)

References 7 publications

(13 reference statements)

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“…mCherry and TagBFP2 were prepared in our previous study . Details of general purification procedures were previously reported. , For expression of msfGFP-MinD, Escherichia coli BL21-CodonPlus(DE3)-RIPL cells (Agilent Technologies, Santa Clara, CA, USA) were transformed with pET15-His-msfGFP-MinD constructed in our previous study and cultivated in LB medium with ampicillin at 37 °C. The protein was expressed by adding 1 mM IPTG at OD 600 = 0.8.…”

Section: Methodsmentioning

confidence: 99%

Controlling the Periodicity of a Reaction–Diffusion Wave in Artificial Cells by a Two-Way Energy Supplier

et al. 2022

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Reaction–diffusion (RD) waves, which are dynamic self-organization structures generated by nanosize molecules, are a fundamental mechanism from patterning in nano- and micromaterials to spatiotemporal regulations in living cells, such as cell division and motility. Although the periods of RD waves are the critical element for these functions, the development of a system to control their period is challenging because RD waves result from nonlinear physical dynamics under far-from-equilibrium conditions. Here, we developed an artificial cell system with tunable period of an RD-driven wave (Min protein wave), which determines a cell division site plane in living bacterial cells. The developed system is based on our finding that Min waves are generated by energy consumption of either ATP or dATP, and the period of the wave is different between these two energy suppliers. We showed that the Min-wave period was modulated linearly by the mixing ratio of ATP and dATP and that it was also possible to estimate the mixing ratio of ATP and dATP from the period. Our findings illuminated a previously unidentified principle to control the dissipative dynamics of biomolecules and, simultaneously, built an important framework to construct molecular robots with spatiotemporal units.

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