Propagation and molecular characterization of avian pneumoviruses
Jacobs, Janet Ashley
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The replication characteristics of Avian pneumovirus (APV) subtype C were studied by generation of a growth curve and by northern blot hybridization of cell-infected RNA with APV N and M genes. APV/C was detected in cell culture media by 24 hours post- infection replicating to a maximum titer of 106 by six days post-infection. APV/C transcription could be detected by northern blot hybridization within 24 hours post- infection and continued for 5 days. The N, P, M, F, and M2 genes for the APV subtypes A, B and C were sequenced. Phylogenetic analysis of the APV N, P, M, F and M2 contiguous sequences confirmed the presence of three APV subgroups. Finally, polyacrylamide gel electrophoresis was performed on purified APV/C proteins to determine if the attachment protein, G, was present since it has yet to be cloned. A putative G protein was detected by Coomassie and glycoprotein staining.