FIG. 1.

Western blot analysis of proteolytic processing of the NS2B-NS4A polyprotein expressed by recombinant baculoviruses. S. frugiperda SF21 cells were infected at a multiplicity of 5 PFU per cell with recombinant baculoviruses or wild-type (wt) Autographa californica nuclear polyhedrosis virus. Cells were harvested at 60 h postinfection. Proteins were separated by SDS-PAGE, electroblotted onto nitrocellulose membranes, and probed with either rabbit antisera raised to HGV synthetic peptides or with the monoclonal anti-HSV tag antibodies. (A) Schematic representation of constructs. (B) Mutations of HGV protease motifs. Phe₁₅₄ of the HCV substrate specificity pocket is indicated by an asterisk and numbered from the N-terminal residue of NS3. (C) Western blot analysis of NS2B/NS4A polyprotein cleavage with an anti-NS2 antibody. Samples were separated on an SDS-PAGE gel (12% polyacrylamide). (D) Western blot analysis of NS2B/NS4A polyprotein cleavage with anti-NS3 polyclonal antibody. Samples were separated on an SDS-PAGE gel (12% polyacrylamide). (E) Western blot analysis of NS2B/NS4A polyprotein cleavage with anti-HSV tag antibody. Samples were separated by SDS-PAGE (18% polyacrylamide) and probed with anti-HSV tag monoclonal antibody. MW, molecular mass markers (kilodaltons).