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

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

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)547-552
Number of pages6
JournalNucleic acids research
Volume19
Issue number3
DOIs
Publication statusPublished - 1991 Feb 11
Externally publishedYes

Fingerprint

RNA
Messenger RNA
Nucleotides
Thermus
Ribonucleosides
Bacteriophage T7
Proteins
Cell-Free System
In Vitro Techniques
Nucleosides
Polyacrylamide Gel Electrophoresis
Bacteria

ASJC Scopus subject areas

  • Genetics

Cite this

Phosphorothioate-containing RNAs show mRNA activity in the prokaryotic translation systems in vitro. / Ueda, Takuya; Tohda, Hideki; Chikazumi, Nobutoshi; Eckstein, Fritz; Watanabe, Kimitsuna.

In: Nucleic acids research, Vol. 19, No. 3, 11.02.1991, p. 547-552.

Research output: Contribution to journalArticle

Ueda, Takuya ; Tohda, Hideki ; Chikazumi, Nobutoshi ; Eckstein, Fritz ; Watanabe, Kimitsuna. / Phosphorothioate-containing RNAs show mRNA activity in the prokaryotic translation systems in vitro. In: Nucleic acids research. 1991 ; Vol. 19, No. 3. pp. 547-552.
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