Ovine adenovirus OAV287 (OAV) is the prototype of a virus group which is phylogenetically distinct from the mastadenoviruses and aviadenoviruses. The genome arrangement of OAV showed that virus-associated (VA) RNA genes were not located between the reading frames for p52/55K and terminal protein as these overlapped. To determine whether VA genes were located elsewhere, several approaches were used. Nuclear extracts containing RNA polymerase III activity were used to transcribe OAV genome fragments in vitro. A product of approximately 120 bp was produced from two widely separated coding regions of the genome. However, when these were subcloned and used as radiolabelled probes to analyse RNA from OAV-infected cells, no VA-like RNA was detected, although late mRNAs that were transcribed from the regions were identified. In addition, 5' radiolabelling of small RNA species in control- and OAV-infected cells followed by gel analysis did not identify candidate VA RNAs. Radiolabelling of proteins in control- and OAV-infected cells showed that there was little preferential translation of viral proteins compared with host polypeptides, in contrast to the situation for adenovirus 5 (Ad5). In addition, the kinetics of OAV infection were slower than observed for human adenoviruses. Collectively, the data suggest that OAV is unique in that it does not produce VA RNA during infection. This conclusion is supported by a comparison of the genomes of the phylogenetically related OAV and egg drop syndrome viruses which shows that the VA gene identified in the latter is located in a region absent from OAV.