The generation and characterization of avian reovirus temperature sensitive mutants

Abstract

Currently, little is known about the genetic and biological functions of avian reovirus (ARV), an atypical member of the family 'Reoviridae ' and the prototype of all non-enveloped syncytia-inducing viruses. In this study, we created temperature sensitive ('ts') mutants by chemical mutagenesis of ARV strain 138. We developed a novel efficiency of lysis (EOL) technique to screen for temperature sensitivity and used the classical efficiency of plating (EOP) assay to identify 17 ARV 'ts ' mutants. Pairwise mixed infection of these mutants and evaluation of recombinant progeny 'ts' status led to their organization into 7 recombination groups. This indicates that these new groups of ARV mutants represent the majority of the ARV genome. To phenotypically characterize each mutant and localize the effects of the 'ts' lesion within the viral lifecycle, the ability to synthesize double stranded RNA (dsRNA) at permissive (33.5C) and non-permissive temperature (39.5C) was analyzed. (Abstract shortened by UMI.)