Abstract
Fowl adenovirus 4 (FAdV-4) is associated with economically important poultry diseases. Recent studies of fully sequenced genomes of FAdV-4 isolates suggest potential genomic regions associated with virulence and amenable for manipulation and vector development. Direct manipulation of viral genomes is cumbersome, as opposed to that of infectious clones—viral genomes cloned into plasmid or cosmid vectors. In this work, we generated an infectious clone, pFAdV-4 ON1, containing the entire viral genome of a nonpathogenic FAdV-4 (ON1 isolate). pFAdV-4 ON1 was used for targeted deletion of open reading frames (ORFs) 16 and 17 and replacement with the enhanced green fluorescence protein (EGFP) expression cassette to generate recombinant viruses. These viruses were viable, and EGFP was expressed in infected cells. Their replication, however, was significantly reduced with respect to that of the wild-type virus. These observations suggest the potential utility of FAdV-4 as a vaccine vector and the importance of ORFs 16 and 17 for virus replication at wild-type levels. To our knowledge, this is the first report of an infectious clone based on the FAdV-4 genome, and our results demonstrate its utility for studies of virulence determinants and as a platform for either vaccine or gene delivery vectors.
Highlights
Fowl adenovirus 4 (FAdV-4) is a member of the species Fowl aviadenovirus C belonging to the genus Aviadenovirus, family Adenoviridae
FAdV-4 is associated with economically important poultry diseases such as inclusion body hepatitis (IBH) and hydropericardium syndrome (HPS) [1,2,3]
One of these isolates is the nonpathogenic FAdV-4 ON1, which was isolated from a Canadian broiler breeder flock with no clinical signs of IBH/HPS [8]
Summary
Fowl adenovirus 4 (FAdV-4) is a member of the species Fowl aviadenovirus C belonging to the genus Aviadenovirus, family Adenoviridae. The virulence determinants of FAdV-4 and the molecular basis of IBH and HPS pathogenesis are unknown. The complete nucleotide sequence has been determined for the genomes of 24 FAdV-4 isolates with nucleotide sequence homologies of 97–99% (see accession numbers GU188428; HE608152; KP295475; KU342001; KU991797; KU587519; KM096544; KX538980; KU558762; KX061750; KU569295; KX090424; KY436520; KU569296; KY436522; KU558761; KY379035; KU558760; KY436521; KY436519; KU245540; KX421403; KX421404; KX421401). One of these isolates is the nonpathogenic FAdV-4 ON1, which was isolated from a Canadian broiler breeder flock with no clinical signs of IBH/HPS [8].
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