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. 2005 Mar 8;33(5):1474–1486. doi: 10.1093/nar/gki292

Figure 3.

Figure 3

Synthesis of the F protein with constructs containing insertions of different length of the HCV-coding sequence. The synthesis of the F protein was measured with constructs containing HCV segments of different length described in Figure 2. In the pHCV-387-LUC, pHCV-426-LUC, pHCV-447-LUC and pHCV-510-LUC cap and IRES constructs, the consensus HCV sequence (AAAGAAAAAC, nt 364–373) occupies the site where a +1 frameshift was proposed to occur. In the 10A corresponding constructs, the consensus HCV sequence is replaced with a 10A stretch. (A) Synthesis of the F protein in cultured cells. Synthesis of the F protein was measured after co-transfection of 293FT cells with 3 μg of a pHCV-LUC (0) or (+1) construct and 1 μg of pcDNA3.1/Hygro(+)/lacZ, which is used to normalize for variations in transfection efficiency. (B) Synthesis of the F protein in vitro. In vitro translation experiments were carried out in 25 μl of RRL with 0.2 μg of mRNAs transcribed from the FbaI-digested pHCV-LUC constructs. Results are reported as the amount of F protein synthesized (assessed by the activity of luciferase in the +1 reading frame) relative to the amount of the polyprotein synthesized (assessed by the activity of luciferase in the 0 reading frame) and were calculated as described in the text. Each value represents the mean ± standard error of four to six independent experiments.