2Institute of Limnology, Siberian Division of the Russian Academy of Sciences, ul. Ulan-Batorskaya 3, Irkutsk 664033, Russia; fax: (3-952) 42-5405
* To whom correspondence should be addressed.
Received December 19, 2003; Revision received February 10, 2004
Recombinant RNA-dependent RNA polymerase of hepatitis C virus was purified using a bacterial expression system (Escherichia coli). The system for enzyme activity detection was optimized. The maximum activity was achieved when the reaction was carried out at 30°C in the presence of 3 mM Mg2+ or 0.75 mM Mn2+. Among alpha- and beta-pyrogallaldehydes, effective inhibitors were found. It was shown that they acted at the primer elongation stage, and their binding to the protein is reversible.
KEY WORDS: hepatitis C virus, RNA-dependent RNA polymerase, non-nucleoside inhibitors