Establishment of a cell-free translation system from rice callus extracts

Kakeru Suzuki, Haruka Inoue, Satoshi Matsuoka, Ryugo Tero, Ayumi Hirano-Iwata, Yuzuru Tozawa

Research output: Contribution to journalArticlepeer-review

Abstract

Eukaryotic in vitro translation systems require large numbers of protein and RNA components and thereby rely on the use of cell extracts. Here we established a new in vitro translation system based on rice callus extract (RCE). We confirmed that RCE maintains its initial activity even after five freeze-thaw cycles and that the optimum temperature for translation is around 20°C. We demonstrated that the RCE system allows the synthesis of hERG, a large membrane protein, in the presence of liposomes. We also showed that the introduction of a bicistronic mRNA based on 2A peptide to RCE allowed the production of two distinct proteins from a single mRNA. Our new method thus facilitates laboratory-scale production of cell extracts, making it a useful tool for the in vitro synthesis of proteins for biochemical studies.

Original languageEnglish
Pages (from-to)2028-2036
Number of pages9
JournalBioscience, Biotechnology and Biochemistry
Volume84
Issue number10
DOIs
Publication statusPublished - 2020 Oct 2

Keywords

  • Cell-free translation
  • Oryza sativa
  • callus
  • in vitro protein synthesis
  • rice

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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