FIG. 9.

Relative gp64 content of wild-type and recombinant baculovirus particles. Sf9 cells were infected at a multiplicity of about 0.01 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. BV progeny were partially purified and solubilized, and total virion proteins were resolved by SDS-PAGE as described in Materials and Methods. Either the gels were stained with Coomassie blue or proteins were transferred to Immobilon filters and probed with a mixture of two different monoclonal antibodies against gp64 and one against AcMNPV capsid as described in Materials and Methods. The relevant proteins in the stained gel were quantitated by using the Gel Doc 1000 system equipped with image analysis software (Molecular Analyst version 2.1; Bio-Rad). (A) Stained gel; (B) Western blot; (C) results of image analysis shown as the gp64/capsid ratio for each type of virus.