TABLE 3.
CFU/μg of HCV RNA obtained with the parental replicon (wild type) and with RNAs carrying given amino acid substitutions in the HCV polyproteina
| HCV protein(s) | Amino acid substitution | Mean CFU/μg of RNA ± SD |
|---|---|---|
| None | None | 40 ± 14 |
| NS3 | 1283 Arg→Gly | 285 ± 60 |
| NS3 | 1383 Glu→Ala | 70 ± 25 |
| NS3 | 1577 Lys→Arg | 20 ± 5 |
| NS3 | 1609 Lys→Glu | 150 ± 65 |
| NS3 | All NS3 mutations | 385 ± 25 |
| NS4B | 1936 Pro→Ser | 500 ± 210 |
| NS5A | 2163 Glu→Gly | 2,650 ± 700 |
| NS5A | 2330 Lys→Glu | 35 ± 5 |
| NS5B | 2442 Ile→Val | 20 ± 10 |
| NS3-5B′ | 9-13F (SfiI fragment) | 4,250 ± 650 |
| NS5B | 2884 Arg→Gly | 23,000 ± 3,000 |
| NS3 + NS5B | 1283 Arg→Gly + 1383 Glu→Ala + 1577 Lys→Arg + 1609 Lys→Glu + 2884 Arg→Gly | 51,000 ± 6,000 |
| NS4B + NS5B | 1936 Pro→Ser + 2884 Arg→Gly | 750 ± 400 |
| NS5A + NS5B | 2163 Glu→Gly + 2884 Arg→Gly | n.a. |
| NS5A + NS5B | 2330 Lys→Glu + 2884 Arg→Gly | 19,000 ± 3,000 |
| NS5B + NS5B | 2442 Ile→Val + 2884 Arg→Gly | 9,000 ± 2,000 |
a
Numbers refer to the amino acid positions of the polyprotein of the HCV consensus genome, which can be retrieved from the EMBL nucleotide sequence database under accession number AJ238799. Values were derived from several independent transfection experiments using serial dilutions of each RNA construct. n.a., not applicable because no or only one colony was obtained in several independent experiments.