TABLE 1.
RNA encapsidation and protection by the nucleocapsidsa
| Mutant | Mean no. of viral RNA copies/5 μl of isolated RNA |
Relative RNA quantification |
||
|---|---|---|---|---|
| Untreated | Treated | No. of copies in untreated NC/no. in treated NC | No. of copies in treated NC/no. in untreated NC (% of wt) | |
| wt | 8.94 × 107 | 4.92 × 105 | 182 | 5.5 × 10−3 (100) |
| wt-SDSb | 6.87 × 107 | 4.26 × 103 | 16,129 | 6.2 × 10−5 (1) |
| R50 mutant | 2.52 × 107 | 1.32 × 105 | 192 | 5.22 × 10−3 (95) |
| K51 mutant | 3.47 × 107 | 1.66 × 105 | 209 | 4.78 × 10−3 (87) |
| R59 mutant | 3.25 × 107 | 3.45 × 105 | 94 | 1.06 × 10−2 (193) |
| R62 mutant | 7.25 × 107 | 4.33 × 105 | 167 | 5.98 × 10−3 (109) |
| JFH1/ΔE1E2 | 5.38 × 107 | 1.36 × 105 | 396 | 2.52 × 10−3 (46) |
a
Core protein-associated RNAs were quantified by qRT-PCR, and the ratios in untreated NC/NC treated with RNase A and DNase and in treated NC/untreated NC were calculated. Ratios of values in treated/untreated NC were compared to the wt ratio, and results are expressed as a percentage of the wt, which was set to 100%. This method enabled us to minimize the differences linked to electroporation efficiency and manipulation. Means of the results from duplicate electroporations are shown.
b
SDS, lysate pretreated with 1% SDS.