Abstract
Abstract Nucleotide polymerization by partially purified f2 replicase in the presence of phage f2 RNA templates is stimulated by salt and by a brief preliminary incubation of the enzyme at high ionic strength. Salt stimulation of enzyme activity is observed with both plus (viral) and minus (complementary) strand RNA templates, is independent of the composition of the salt, and is maximal at an ionic strength of 0.1. The polynucleotide products of the in vitro reaction are homologous to f2 RNAs by annealing criteria. The in vitro product made in the presence of a mixture of plus and minus strand RNAs consists largely of f2 plus strand RNA. Both plus and minus strand RNAs are synthesized in the presence of plus strand template. Less than 20% of the replicase product sediments as 27 S single-stranded viral RNA in glycerol gradients. Most of the in vitro product is present in partially double-stranded species sedimenting in a broad band at about 15 S. These results suggest that the f2 replicase is capable of limited in vitro chain termination and reinitiation of RNA synthesis.
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