TABLE 2.
Virus replication assay
| Virus | Genotype of of UL15 | Cell line infected/cell line used to determine yielda | Titerb |
|---|---|---|---|
| HSV-1(F) | WT | Vero/Vero | (2.0 ± 0.55) × 107 |
| HSV-1(F) | WT | CV15/Vero | (2.0 ± 0.4) × 107 |
| HSV-1(F) | WT | CV15M/Vero | (2.5 ± 0.6) × 105 |
| 15 null | Δ exon IIc | CV15/CV15 | (1.0 ± 0.5) × 107 |
| 15 null | Δ exon II | CV15M/CV15 | <102 |
| vJB12 | ΔNLS | CV15/CV15 | (6.1 ± 1.5) × 105 |
| vJB12 | ΔNLS | Vero/CV15 | <102 |
| vJB13 | FLAG tag at C terminus | Vero/Vero | (2.0 ± 0.2) × 107 |
| vJB15 | NLS repaired (derived from vJB12) | Vero/Vero | (2.0 ± 1.5) × 107 |
| vJB16 | FLAG tag at C terminus, ΔNLS (derived from vJB13) | Vero/CV15 | <102 |
| vJB19 | FLAG tag at C terminus, NLS repaired (derived from vJB16) | Vero/Vero | (2.2 ± 1.5) × 107 |
a
The first listed cell line was infected at 0.01 PFU/cell, and at 24 hpi, the yield of infectious virus produced from pooled extracellular and intracellular fractions was determined on the second listed cell line. CV15 is an engineered complementing cell line expressing wild-type (WT) pUL15, whereas CV15M was engineered to express pUL15 lacking the NLS by deletion of codons 182 to 189.
b
Virus titers were determined as described in Materials and Methods. The data represent means ± standard deviations for three independent experiments.
c
Delta (Δ) indicates a deletion of those sequences.