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

Yukino Matsui; Toshiki Akui; Nobuhide Doi; Kei Fujiwara

doi:10.1016/j.xpro.2021.101003

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

STAR Protoc. 2021 Dec 11;2(4):101003. doi: 10.1016/j.xpro.2021.101003. eCollection 2021 Dec 17.

Authors

Yukino Matsui¹, Toshiki Akui¹, Nobuhide Doi¹, Kei Fujiwara¹

Affiliation

¹ Department of Biosciences & Informatics, Keio University, 3-14-1 Hiyoshi, Kohokuku, Yokohama, Kanagawa 223-8522, Japan.

Abstract

We present a protocol for activating protein synthesis in liposomes encapsulating a diluted E. coli cell extract-based TX-TL (transcription-translation) system by hypertonic concentration. Protein expression is turned on in the liposome-encapsulated TX-TL system by simple treatment with a concentrated external solution. The expression of sfGFP is demonstrated here, but it can be applied to other proteins. This protocol can be applied to the development of artificial cells utilizing the switch-on mechanism to activate protein expression, responding to the outer environment. For complete details on the use and execution of this protocol, please refer to Akui et al. (2021).

Keywords: Biophysics; Biotechnology and bioengineering; Molecular Biology; Protein Biochemistry.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell-Free System
Escherichia coli / metabolism*
Liposomes*
Osmolar Concentration
Protein Biosynthesis*
Transcription, Genetic*

Substances

Liposomes