FIG. 5.

Expression of gp64 N-glycosylation site mutants by recombinant baculoviruses. Sf9 cells were infected at a multiplicity of 2.5 PFU per cell with wild-type AcMNPV (WT) or recombinant baculoviruses encoding mutant gp64s lacking various consensus N-glycosylation sites, as indicated above the lanes. The cells were treated with 0.2 mM castanospermine from 1 to 44 h and labeled with 100 μCi of Tran³⁵S-label per ml of radiolabeling medium containing 0.2 mM castanospermine from 44 to 48 h postinfection. At the end of the labeling period, intracellular gp64 was extracted and immunoprecipitated, and the washed immunoprecipitates were analyzed by SDS-PAGE and autoradiography.