FIG. 1.
Construction and characterization of a ΔBLLF1 recombinant virus. (A) Schematic map of the EBV-wt and the ΔBLLF1 genomes generated by exchange of the BLLF1 gene against the kanamycin resistance cassette (kanr). The regions of homology between the targeting vector and the wild-type genome are indicated (hatched line). The figure includes the cleavage sites for XhoI (X) and the expected fragment sizes after restriction analysis of EBV-wt and ΔBLLF1 genomes. (B) XhoI restriction fragment analysis of EBV-wt (lane 1) and ΔBLLF1 mutant genomes after construction in E. coli (lane 2) and rescue from 293/ΔBLLF1 cells (lane 3). The result is consistent with the predicted restriction pattern (see panel A). (C) Gp350 protein production was examined by immunostaining of induced 293/ΔBLLF1 and 293/ΔBLLF1-C cells with a gp350-specific antibody. (D) Detection of gp350 within purified viral particles produced by 293/ΔBLLF1 (left) and 293/ΔBLLF1-C (right) by Western blot analysis with a gp350-specific antibody.