FIG. 2.

Expression of the ORF 3 protein in infected insect cells. Sf9 cells were either mock infected (Mock) or infected with wild-type baculovirus (WT), the ORF 2 baculovirus recombinant, the ORFs 2+3 baculovirus recombinant, or the ORF 3 baculovirus recombinant. (A) Kinetics of ORF 3 expression in infected insect cells. Cells were harvested at 30, 48, and 72 h p.i. by lysis in SDS-PAGE sample buffer. The samples were separated by SDS-PAGE in a 15% polyacrylamide gel, and Western blot analysis was performed using the rabbit ORF 3 C-terminal peptide antiserum at a dilution of 1:500. The asterisk highlights the 23K ORF 3 protein band that separates into two bands on some gels. (B) Immunoprecipitation of the ORF 3 protein from infected insect cells. Cells infected with wild-type baculovirus (WT), the ORF 3 or the HA-tagged ORF 3 (ORF 3-HA) baculovirus recombinants were harvested in RIPA buffer at 65 h p.i. and immunoprecipitated using the ORF 3 C-terminal peptide antiserum.