Establishment of a cell-free translation system from rice callus extracts.
Biosci Biotechnol Biochem
; 84(10): 2028-2036, 2020 Oct.
Article
em En
| MEDLINE
| ID: mdl-32543982
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.
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MEDLINE
Assunto principal:
Oryza
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Biossíntese de Proteínas
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Extratos Vegetais
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article