Assessment of sequence relatedness of double-stranded RNA genes by RNA-RNA blot hybridization

Abstract

Three well-characterized reovirus serotypes were used to investigate the usefulness of RNA-RNA blot hybridization as a means to assess the genetic relatedness of double-stranded RNA (dsRNA) viruses. [5'- 32P]pCp-labeled genomic dsRNAs from reovirus 1, 2 and 3 were used as probes in hybridization experiments in which segments of the three serotypes were separated in 10% polyacrylamide gels and transferred electrophoretically to membranes. Nine of the 10 reovirus genes cross-hybridized between the serotypes. The S1 gene was serotype specific. The L2 gene of reovirus 2 showed a lower level of cross-hybridization with types 1 and 3 when compared to the hybridization signal observed for L2 when types 1 and 3 were hybridized to each other. The data were consistent with previous studies on the relatedness of the three virus serotypes. Since RNA-RNA blot hybridization allows the number and identity of conserved genes to be determined, this approach may prove useful for assessing the genetic relatedness among other viruses in the family Reoviridae.