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. Author manuscript; available in PMC: 2014 Apr 25.
Published in final edited form as: Virology. 2013 Feb 26;439(1):23–33. doi: 10.1016/j.virol.2013.01.019

Figure 1. Gene insertion between NS5A and NS5B does not impair viral fitness.

Figure 1

(a) Genome structure of the Jc1(Δ34-378-YPet) and Jc1 5AB YPet illustrating the insertion of YPet either as a fusion protein with NS5A with a subsequent adaptive deletion in domain II (II) or by duplicating the NS3-4A protease cleavage site between NS5A and NS5B (5A/B CS) flanking the YPet insertion. (b) HCV replication following electroporation of in vitro transcribed RNA of Jc1, Jc1(Δ34-378-YPet) or Jc1 5AB YPet into Huh-7.5 cells as measured by flow cytometry. (c) Longitudinal virus production as measured by end-point limiting dilution following electroporation of Jc1, Jc1(Δ34-378-YPet) or Jc1 5AB YPet RNA into Huh-7.5 cells. Results shown are means ± SD from three independent experiments (b, c). (d, e) Histograms of (d) NS5A- and (e) YPet expression following electroporation of Huh-7.5.1 cells with Jc1, Jc1(Δ34-378-YPet)), or Jc1 5AB YPet. (f) NS5A/5B cleavage at different times post electroporation of Jc1, Jc1(Δ34-378-YPet) or Jc1 5AB YPet into Huh-7.5 cells as measured by Western blot against either NS5A or NS5B.