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Full-Length Complementary DNA of Hepatitis C Virus Genome From
an Infectious Blood Sample
Fig. 2. Construction of a
full-length cDNA clone of HCV. (A) Sequences of primers and anchors
for RT-nested PCR. The synthetic oligonucleotide primers are
according to the published nucleotide sequences of type Ib HCV
clones and a novel 98-nt sequence in the 3' terminus. The anchors
(anchor 1, anchor 2') and the anchors' primers (As1, La1', La2')
are obtained from the 5'-Ampli FINDER RACE kit and Marathon cDNA
amplification kit. An oligo RNA (anchor 2') and primers (Ls1, Ls2,
Ls3, Ls4, La1, La2) were according to Tanaka et al.63
(B) Gene organization of the HCV. Shaded and open boxes indicate
structural and nonstructural proteins, respectively. UTRs are
indicated by bars. Black and white triangles indicate cleavage
sites by cellular signalase and viral-coded proteases,
respectively. (C) Strategy for making a full-length cDNA clone of
HCV. The locations of primers and anchors are shown. The 5' and 3'
terminal sequences were determined by use of a 5'-Ampli FINDER RACE
kit and an RNA linker ligation, respectively.
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