The Mutant Distribution of an RNA Species Replicated by Qβ Replicase

The mutant spectrum of an RNA species that is replicated by Qβ replicase, MNV-11, was investigated by retrotranscribing the RNA to DNA and cloning it into plasmids. The sequences of several cDNA clones of MNV-11 populations amplified by Qβ replicase under various conditions were determined and compa...

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Veröffentlicht in:Journal of molecular biology 1995-06, Vol.249 (4), p.754-762
Hauptverfasser: Rohde, Nicole, Daum, Heidrun, Biebricher, Christof K.
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Sprache:eng
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Zusammenfassung:The mutant spectrum of an RNA species that is replicated by Qβ replicase, MNV-11, was investigated by retrotranscribing the RNA to DNA and cloning it into plasmids. The sequences of several cDNA clones of MNV-11 populations amplified by Qβ replicase under various conditions were determined and compared. A surprisingly broad mutant distribution was found: the consensus sequence never made up more than 40% of the total population and was accompanied by many mutants. Most mutants had several base exchanges, insertions and/or deletions; up to nine of the total 86 nucleotides were changed. The mutants found had replication rates comparable to that of the wild-type and were thus enriched in the population by selection forces. When the growth conditions were changed, the mutant distribution centre was shifted. The published consensus sequence of MNV-11 did not have the highest growth rate of the mutants, but was rather the best adapted to the various selection forces governing the growth phases the replicating RNA went through, i.e. it had found an optimal compromise between the rates of overall replication, enzyme binding and double strand formation.f2f2Abbreviations used: TGGE, temperature gradient gel electrophoresis; HD, Hamming distance.
ISSN:0022-2836
1089-8638
DOI:10.1006/jmbi.1995.0334