Synergism in replication and translation of messenger RNA in a cell-free system

I.Y. Morozov, V.I. Ugarov, A.B. Chetverin, A.S. Spirin

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)

Abstract

Combination of the Q beta replicase reaction with the Escherichia coli cell-free translation system markedly enhances replication of a recombinant RQ-DHFR RNA consisting of the dihydrofolate reductase (DHFR) mRNA sequence inserted into RQ135(-1) RNA, an efficient naturally occurring Q beta replicase template. The enhancement is associated with a replication asymmetry previously described for the replication of Q beta phage RNA in vivo; the sense (+)-strands are produced in large excess over the antisense (-)-strands. This, in turn, results in increased synthesis of the functionally active DHFR. These effects are not observed when DHFR mRNAs or RQ135(-1) RNAs are used as templates, if the translation system is not complete, or if it is inhibited by puromycin. The coupled replication-translation of nonviral mRNA recombinants can serve as a useful model for studying the fundamental aspects of virus amplification and can be implemented for large-scale protein synthesis in vitro.
Original languageEnglish
Pages (from-to)9325-9329
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number20
DOIs
Publication statusPublished - 15 Oct 1993
Externally publishedYes

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