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 hepatitis. Since it has been difficult to obtain full-length cDNA clones of HCV including the 3' untranslated region (UTR) that give rise to replication competent virus, we generated the 3'UTR by a modified protocol of total chemical synthesis (TCS) with overlap-extension-PCR using four long oligodeoxynucleotides. A synthetic cDNA fragment of about 340 nucleotides (nt) in length was generated, subcloned and sequenced. This approach represents a rapid and easy alternative to RT-PCR from infectious serum and may be a highly valuable method to generate partial cDNA clones of HCV and other viruses including defined variants.
Encke
J
J
Molecular Hepatology Laboratory, Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown 02129, USA.
zu Putlitz
J
J
Heintges
T
T
Wands
J R
JR
D020413:3' Untranslated Regions / Q000138:chemical synthesis*; D001483:Base Sequence; D004279:DNA, Viral / Q000138:chemical synthesis*; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides
D020413:3' Untranslated Regions; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides