Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids

Shunshi Kohyama; Kei Fujiwara; Natsuhiko Yoshinaga; Nobuhide Doi

doi:10.21769/BioProtoc.3561

Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids

Shunshi Kohyama, Kei Fujiwara, Natsuhiko Yoshinaga, Nobuhide Doi

Department of Biosciences and Informatics

Research output: Contribution to journal › Article › peer-review

Abstract

The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling between the reactions among MinD, MinE, and ATP and the differences in diffusion rate on the cell membrane and in the cytoplasm. Because Min waves appear only under the balanced condition of the reaction-diffusion coupling, special attentions are needed towards several technical points for the reconstitution of Min waves in artificial cells. This protocol describes a technical method for stably generating Min waves in artificial cells.

Original language	English
Article number	e3561
Journal	Bio-protocol
Volume	10
Issue number	6
DOIs	https://doi.org/10.21769/BioProtoc.3561
Publication status	Published - 2020 Mar 20

Keywords

Artificial cells
Cell division
Cell-sized space
In vitro reconstitution
Min system
Reaction-diffusion coupling
Spatiotemporal regulation

ASJC Scopus subject areas

Neuroscience(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)
Plant Science

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10.21769/BioProtoc.3561

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title = "Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids",

abstract = "The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling between the reactions among MinD, MinE, and ATP and the differences in diffusion rate on the cell membrane and in the cytoplasm. Because Min waves appear only under the balanced condition of the reaction-diffusion coupling, special attentions are needed towards several technical points for the reconstitution of Min waves in artificial cells. This protocol describes a technical method for stably generating Min waves in artificial cells.",

keywords = "Artificial cells, Cell division, Cell-sized space, In vitro reconstitution, Min system, Reaction-diffusion coupling, Spatiotemporal regulation",

author = "Shunshi Kohyama and Kei Fujiwara and Natsuhiko Yoshinaga and Nobuhide Doi",

note = "Funding Information: Research was funded by JSPS KAKENHI Grant Number JP16H00809, JP26650044, JP15KT0081, JP15H00826, JP18H04565 (to KF), JP26800219, JP16H00793, and JP17K05605 (to NY). We are also grateful for a Ph.D. Program Research Grant at Keio university awarded to SK. The protocol described here is based on research previously published in Kohyama et al. (2019). Publisher Copyright: Copyright {\textcopyright} 2020 The Authors; exclusive licensee Bio-protocol LLC.",

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doi = "10.21769/BioProtoc.3561",

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AU - Yoshinaga, Natsuhiko

AU - Doi, Nobuhide

N1 - Funding Information: Research was funded by JSPS KAKENHI Grant Number JP16H00809, JP26650044, JP15KT0081, JP15H00826, JP18H04565 (to KF), JP26800219, JP16H00793, and JP17K05605 (to NY). We are also grateful for a Ph.D. Program Research Grant at Keio university awarded to SK. The protocol described here is based on research previously published in Kohyama et al. (2019). Publisher Copyright: Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.

PY - 2020/3/20

Y1 - 2020/3/20

N2 - The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling between the reactions among MinD, MinE, and ATP and the differences in diffusion rate on the cell membrane and in the cytoplasm. Because Min waves appear only under the balanced condition of the reaction-diffusion coupling, special attentions are needed towards several technical points for the reconstitution of Min waves in artificial cells. This protocol describes a technical method for stably generating Min waves in artificial cells.

AB - The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling between the reactions among MinD, MinE, and ATP and the differences in diffusion rate on the cell membrane and in the cytoplasm. Because Min waves appear only under the balanced condition of the reaction-diffusion coupling, special attentions are needed towards several technical points for the reconstitution of Min waves in artificial cells. This protocol describes a technical method for stably generating Min waves in artificial cells.

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KW - Cell-sized space

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Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids

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Keywords

ASJC Scopus subject areas

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