Figure 5

Replication of unmodified sub-genomic HCV 2a RNA clone and GFP labeled sub-genomic HCV 2a chimera in Huh-7 cells. Huh-7 cells were transfected with 10 μg of in vitro transcribed RNA by the electroporation method and then cultured in the medium containing G-418 (500 μg/ml). After 4-weeks, G-418 resistant cell colonies were stained with Giemsa Stain (Sigma Chemical). Both the unmodified (pSGR) and GFP tagged sub-genomic clone (pSGR-GFP) replicated at equal efficiencies based on the development of number of G-418 resistant Huh7 cell colonies. No G-418 resistant colonies were present in Huh-7 cells with GND mutant sub-genomic RNA with GFP (pSGR-GND-GFP).