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.
Ewen
M E
ME
Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637.
Revel
H R
HR
D001435:Bacteriophages / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D066298:In Vitro Techniques; D011549:Pseudomonas; D012330:RNA, Double-Stranded / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D014158:Transcription, Genetic; D014779:Virus Replication
D012330:RNA, Double-Stranded; D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases