Identification of a 24 kDa protein expressed by chicken anaemia virus

Antisera raised against oriented peptide conjugates were used to identify and partially characterize a 24 kDa protein product expressed by chicken anaemia virus (CAV). The peptides derived from the N and C termini of the protein were shown to react against the native protein, expressed within virus-infected cells, by immunofluorescence, immunoperoxidase and immunogold thin section electron microscopy techniques. The protein product was located by immunogold single labelling in intranuclear inclusions similar to those described previously for the 13 kDa CAV protein, which causes apoptosis. The 24 kDa protein was co-localized to the nuclear inclusions with the CAV 13 kDa protein by simultaneous dual labelling immunogold electron microscopy. Following isolation of the CAV proteins by nuclei isolation and SDS-PAGE, the antisera were used to probe for the protein by immunoblotting. The antisera recognized an expressed protein product of apparent molecular mass 30 kDa. An immunofluorescence time course study of CAV protein expression was carried out and the peptide antisera reacted against the protein at 12 h post-infection. Antisera against the 13 kDa protein reacted at similar times post-infection. This was in contrast to antisera raised against the 52 kDa capsid protein which is detectable by immunofluorescence only after 24 h. The 13 kDa and 24 kDa proteins thus appear to be early antigens produced by CAV during infection.