We examined the transcriptional organization of fowl adenovirus 9 (FAdV-9) and analyzed temporal transcription profiles of its early and late mRNAs. At least six early and six late transcriptional regions were identified for FAdV-9. Extensive splicing was observed in all FAdV-9 early transcripts examined. Sequence analysis of the cDNAs representing the early proteins identified untranslated leader sequences, precise locations of splice donor and acceptor sites, as well as polyadenylation signals and polyadenylation sites. A unique characteristic, compared to other adenoviruses, was the detection by RT-PCR of multiple transcripts specific for each of five late genes (protein III, pVII, pX, 100K, and fiber), suggesting that FAdV-9 late transcripts undergo more extensive splicing than reported for other adenoviruses.