ON THE NATURE OF SPONTANEOUS RNA-SYNTHESIS BY Q-BETA-REPLICASE

Numerous RNA species of different length and nucleotide sequence grow spontaneously in vitro in Qβ replicase reactions where no RNA templates are added deliberately. Here, we show that this spontaneous RNA synthesis by Qβ replicase is template directed. The immediate source of template RNA can be the laboratory air, but there are ways to eliminate, or at least substantially reduce, the harmful effects of spontaneous synthesis. Solitary RNA molecules were detected in a thin layer of agarose gel containing Qβ replicase, where they grew to form colonies that became visible upon staining with ethidium bromide. This result provides a powerful tool for RNA cloning and selection in vitro. We also show that replicating RNAs similar to those growing spontaneously are incorporated into Qβ phage particles and can propagate in vivo for a number of phage generations. These RNAs are the smallest known molecular parasites, and in many aspects they resemble both the defective interfering genomes of animal and plant viruses and plant virus satellite RNAs. © 1991.