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

Bio Protoc. 2020 Mar 20;10(6):e3561. doi: 10.21769/BioProtoc.3561.

Authors

Shunshi Kohyama¹, Kei Fujiwara¹, Natsuhiko Yoshinaga^{2

3}, Nobuhide Doi¹

Affiliations

¹ Department of Biosciences and Informatics, Keio University, Yokohama, Japan.
² Mathematical Science Group, WPI Advanced Institute for Materials Research (WPI-AIMR), Tohoku University, Sendai, Japan.
³ MathAM-OIL, AIST, Sendai, Japan.

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.