Figure 1

Construction of CIC by overlapping PCR. During the first-round of PCR, 3 fragments (A, B and C: VP62 33-486 nt, VP62 486-1149, and the HCV 5'UTR) were amplified from plasmid VP62 (kindly provided by Lombardi) and pNCCL-HCV (constructed by our laboratory) using primers Gag-1S and Gag-1A, Gag-2S and Gag-2A, and HCV-S and HCV-A. In the first overlapping PCR, fragment D was amplified from fragments A and B using primers Gag-1S and Gag-2A; the internal probe-binding sequences were introduced into fragment D. Fragment CIC was obtained by a second overlapping PCR using primers Gag-1S and HCV-A to amplify from fragments D and C.