Phosphorothioate-containing RNAs show mRNA activity in the prokaryotic translation systems in vitro

Takuya Ueda*, Hideki Tohda, Nobutoshi Chikazumi, Fritz Eckstein, Kimitsuna Watanabe


研究成果: Article査読

46 被引用数 (Scopus)


Phosphorothioate-containing RNAs were generated by transcription of coliphage T7 ONA using the Sp diastereomers of ribonucleoside 5′-O-(1-thiotriphos-phates) and T7 RNA polymerase. RNAs In which a single nucleotide was substituted by the corresponding nucleoside phosphorothioate functioned as mRNA in the cell-free translation systems prepared from Escherichla coli and from an extreme theimophilic bacterium, Thermus thermophllus. This substitution increased the efficiency of protein synthesis by stabilizing the mRNAs in these systems. As the proportion of substituted nucleotides was increased, their mRNA activity was decreased accordingly. As judged from the analysis by SDS-polyacrylamide gel-electrophoresis, the proteins synthesized using phosphorothioate-containing mRNAs as template were identical to those obtained with unsubstituted mRNAs. However, larger proteins which were barely detectable when unsubstituted mRNA was used were well represented when phosphorothioate-RNA was used instead. The advantages in using the phosphorothioate-mRNAs in the in vitro translation systems are discussed.

ジャーナルNucleic acids research
出版ステータスPublished - 1991 2月 11

ASJC Scopus subject areas

  • 遺伝学


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