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 substitutionMean CFU/μg of RNA ± SD
NoneNone40 ± 14
NS31283 Arg→Gly285 ± 60
NS31383 Glu→Ala70 ± 25
NS31577 Lys→Arg20 ± 5
NS31609 Lys→Glu150 ± 65
NS3All NS3 mutations385 ± 25
NS4B1936 Pro→Ser500 ± 210
NS5A2163 Glu→Gly2,650 ± 700
NS5A2330 Lys→Glu35 ± 5
NS5B2442 Ile→Val20 ± 10
NS3-5B′9-13F (SfiI fragment)4,250 ± 650
NS5B2884 Arg→Gly23,000 ± 3,000
NS3 + NS5B1283 Arg→Gly + 1383 Glu→Ala + 1577 Lys→Arg + 1609 Lys→Glu + 2884 Arg→Gly51,000 ± 6,000
NS4B + NS5B1936 Pro→Ser + 2884 Arg→Gly750 ± 400
NS5A + NS5B2163 Glu→Gly + 2884 Arg→Glyn.a.
NS5A + NS5B2330 Lys→Glu + 2884 Arg→Gly19,000 ± 3,000
NS5B + NS5B2442 Ile→Val + 2884 Arg→Gly9,000 ± 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.