Your browser doesn't support javascript.
loading
Establishment of a cell-free translation system from rice callus extracts.
Suzuki, Kakeru; Inoue, Haruka; Matsuoka, Satoshi; Tero, Ryugo; Hirano-Iwata, Ayumi; Tozawa, Yuzuru.
Afiliação
  • Suzuki K; Graduate School of Science and Engineering, Saitama University , Saitama, Japan.
  • Inoue H; Graduate School of Science and Engineering, Saitama University , Saitama, Japan.
  • Matsuoka S; Graduate School of Science and Engineering, Saitama University , Saitama, Japan.
  • Tero R; Department of Applied Chemistry and Life Science, Toyohashi University of Technology , Toyohashi, Japan.
  • Hirano-Iwata A; Advanced Institute for Materials Research, Tohoku University , Sendai, Japan.
  • Tozawa Y; Graduate School of Science and Engineering, Saitama University , Saitama, Japan.
Biosci Biotechnol Biochem ; 84(10): 2028-2036, 2020 Oct.
Article em En | MEDLINE | ID: mdl-32543982
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.
Assuntos
Palavras-chave

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Oryza / Biossíntese de Proteínas / Extratos Vegetais Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Oryza / Biossíntese de Proteínas / Extratos Vegetais Idioma: En Ano de publicação: 2020 Tipo de documento: Article