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

Yukino Matsui, Toshiki Akui, Nobuhide Doi, Kei Fujiwara

Research output: Contribution to journalArticlepeer-review

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).

Original languageEnglish
Article number101003
JournalSTAR Protocols
Volume2
Issue number4
DOIs
Publication statusPublished - 2021 Dec 17

Keywords

  • Biophysics
  • Biotechnology and bioengineering
  • Molecular Biology
  • Protein Biochemistry

ASJC Scopus subject areas

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

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