Figure 1. .

Construction and characterization of the HSV Us9 mutants. (A) Schematic of the HSV unique short (Us9) component. Most of the Us9 gene was deleted in F Us9-. The first 17 nts were intact and the final 205 to 170 nts were frame shifted (gray). The Us9R strain contained the reintroduced deleted sequences. In the Us9-30 strain, nine amino acids were deleted, and remaining portions of the gene were intact. The NS strains included the wt strain and a mutant strain, Us9-, in which the entire Us9 gene was replaced by a mCherry marker. (B) Protein expression. Vero cells were infected with either F strain Us9R, Us9-, or Us9-30 or NS strain wt or Us9- using 1 pfu/cell for 24 hours. Antibodies directed against gB, gE, gD, VP5, or Us9 were used to probe for protein expression. Us9 was not detected in F Us9- or in NS Us9- infected cells. However, Us9 was detected in Us9-30 infected cells at a slightly lower molecular weight location. Note that one of the higher molecular weight bands of Us9 in the Us9-30 appears to be reduced. Molecular markers are 100 (gB), 70 (gE), 58 (gD), 155 (VP5), and 17 (Us9) kDa.