FIG. 1.

Construction of ie1 knockout, rescue, and WT AcMNPV bacmids. (A) Schematic maps of the AcMNPV ie1 gene locus showing the relative positions and orientations of the orf146, ie1, and odv-e56 ORFs (top) and recombination vector plasmid pAcIE1(fr)-Zeo (bottom). pAcIE1(fr)-Zeo contains the ie1 ORF replaced by the Zeocin resistance-encoding gene and was used to generate the ie1 knockout bacmid by recombination in E. coli. (B) Schematic diagrams of rescue viruses AcBacIE1KO, AcBacSplice, AcBacIE1, AcBacIE1(rev), AcBacIE0, and AcBacIE0 showing the genes inserted into the polyhedrin (polh) locus by Tn7-mediated transposition. (C) Western blot analysis for the presence or absence of IE1 or IE0 protein expressed from bacmids AcBacIE1KO, AcBacWT, AcBacSplice, AcBacIE1, AcBacIE1(rev), AcBacIE0, and AcBacIE0. Locations of the IE0- and IE1-specific bands are shown on the right.