In vitro replication and transcription of the segmented double-stranded RNA bacteriophage phi 6.

In vitro conditions that support viral-specific replication and transcription have been developed from Pseudomonas phaseolicola cells infected with the segmented double-stranded RNA bacteriophage phi 6. Transcription activity, previously shown to occur by semiconservative strand displacement, labeled (+) strands of all three genome segments and produced all three corresponding genome length messenger RNAs. Replication activity for each of the three double-stranded RNA segments is observed. Our criteria for replication were formation of genomic length double-stranded RNA products and at least (-) strand synthesis activity. Mn2+ and Sarkosyl together selectively inhibited transcription. Analysis of replication alone suggested that replication templates are the viral (+) messenger RNAs.