TOTAL CHEMICAL SYNTHESIS OF THE 3'-UNTRANSLATED REGION OF THE HEPATITIS-C VIRUS WITH LONG OLIGODEOXYNUCLEOTIDES

Hepatitis C Virus (HCV) is the major causative agent of chronic hepati tis. Since it has been difficult to obtain full-length cDNA clones of HCV including the 3' untranslated region (UTR) that give rise to repli cation competent virus, we generated the 3'UTR by a modified protocol of total chemical synthesis (TCS) with overlap-extension-PCR using fou r long oligodeoxynucleotides. A synthetic cDNA fragment of about 340 n ucleotides (nt) in length was generated, subcloned and sequenced. This approach represents a rapid and easy alternative to RT-PCR from infec tious serum and may be a highly valuable method to generate partial cD NA clones of HCV and other viruses including defined variants. (C) 199 8 Elsevier Science B.V. All rights reserved.