FIG. 1.
Identification of the ORF4-encoded protein. ORF4 RNA transcripts were translated in vitro (ivt) in the absence or presence of canine pancreatic microsomes, and the expression products were immunoprecipitated with the GP4-specific antipeptide serum (GP4E) or its preimmune serum (pGP4E). The precipitated proteins were treated or mock treated with PNGase F or endo H. For comparison, EAV-infected BHK-21 C13 cells were pulse-labeled for 15 min with [35S]cysteine at 8.25 h p.i. and chased for 30 min. Cells were lysed, and IPs were performed with the GP4-specific antipeptide serum (EAV). The precipitated proteins were also treated or mock treated with PNGase F or endo H. All samples were dissolved in LSB containing 5% β-mercaptoethanol and analyzed in SDS-15% PAA gels. The values on the left are the molecular sizes, in kilodaltons, of marker proteins analyzed in the same gel.