Molecular cloning and restriction enzyme mapping of avian adenovirus type 8 DNA

Avian adenovirus (AAV) type 8 was cultured in an avian hepatoma cell line designated CH-SAH and the viral DNA extracted and purified. Restriction enzyme analysis of viral DNA using the endonucleases ApaI, EcoRI, HindIII, KpnI, NotI, SpeI, StuI and XbaI was carried out, and fragments representing the entire genome were cloned. According to the restriction enzyme fragments, the size of the AAV type 8 genome was calculated to be 44.7 kb. Subcloning of viral DNA fragments and hybridization studies using selected viral DNA fragments facilitated the construction of the physical map of AAV type 8 DNA.