FIG. 6.
Impact of additional adaptive mutations on replication competence of the subgenomic genotype 1a replicon, Htat2ANeo/QR/KR/SI (see Fig. 4). (A) Location of various adaptive mutations within the second open reading frame (derived entirely from the genotype 1a H77sequence): Q1067R and V1655I (NS3); K1691R (NS4A); and K2040R (KR5A), F2080V, and S2204I (NS5A). Detailed information concerning each mutation is described in the text. (B) Transient HCV RNA replication assay. SEAP activity in culture supernatants collected at 24 h intervals following electroporation of En5-3 cells with the 1a replicon Htat2ANeo carrying the indicated combinations of the adaptive mutations shown in panel A. Cells were also transfected with genotype 1b Bpp-Ntat2ANeo/SI replicon RNA as a reference. See legend to Fig. 2 for further details. See Table 1 for identification of adaptive mutations in each transcript.