Figure 2
Characterization of HycuMNPV bacmid (HycuBac) in SpIm cells. (a) Schematic representation of HycuBac. (b) Microscopic images of SpIm cells transfected with pIE1-2/Egfp and HycuBac. SpIm cells were transfected with 2 μg of pIE1-2/Egfp, which expresses EGFP protein, or HycuBac and examined at 120 h post-transfection. Scale bar, 50 μm. (c) Microscopic images of virus-infected SpIm cells. mock, Mock-infected SpIm cells. Scale bar, 50 μm. (d) BV yields in virus-infected SpIm cells. BV titers were determined by a plaque assay using SpIm cells. The vertical bars represent the standard deviations of the averages from three determinations. (e) Immunoblot analysis of polyhedrin (Polh) protein in virus-infected SpIm cells. Polyhedrin was probed using anti-BmNPV polyhedrin polyclonal antibody and visualized with a HRP Conjugate Substrate Kit. mock, Mock-infected SpIm cells. Polypeptides equivalent to 1 × 105 cells were analyzed in each lane. In (c), (d) and (e), SpIm cells were infected with HycuMNPV or HycuMNPV derived from HycuBac-transfected SpIm cells (vHycuBac) at an MOI of 10 and examined at 0, 24, 48 and 72 h post-infection (pi).
