FIG. 1.

Determination of the nucleotide sequences at the 5′-and 3′ ends of HCV RNA produced by the Pol I system. (A and B) 5′RACE and sequence analysis. A synthesized RNA adapter was ligated to RNA extracted from cells transfected with pHHJFH1. The positive-strand HCV RNA was reverse transcribed, and the resulting cDNA was amplified by nested PCR. The amplified 5′-end cDNA was separated by agarose gel electrophoresis (A), cloned, and sequenced (B). (C and D) 3′RACE and sequence analysis. RNA extracted from pHHJFH1-transfected cells, the culture supernatant of transfected cells, and the culture supernatant of H751JFH1/Zeo cells were polyadenylated, reverse transcribed, and amplified by PCR. The amplified 3′-end cDNA was separated by agarose gel electrophoresis (C), cloned, and sequenced (D).