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J Gen Virol 76 (1995), 3009-3019; DOI 10.1099/0022-1317-76-12-3009
© 1995 Society for General Microbiology
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Recombinant baculovirus-expressed NS3 proteinase of hepatitis C virus shows activity in cell-based and in vitro assays

Hilary Overton*, David McMillan, Fiona Gillespie and John Mills

Department of Molecular Virology, Roche Products Ltd, Welwyn Garden City AL7 3AY, UK

Recombinant baculoviruses have been constructed which express the hepatitis C virus (HCV) NS3 proteinase and its substrates in insect cells. The expressed proteinase has been shown to carry out trans-cleavage at the NS3/4A, NS4A/4B, NS4B/5A and NS5A/5B junctions in a cell-based assay. When assayed in a cell-free system using in vitro translated substrates, the proteinase could perform Trans-processing of the NS4A/4B and NS5A/5B junctions, but only when coexpressed with NS4A, either as an NS3-4A precursor or by co-infection of cells with NS3- and NS4A-expressing recombinant baculoviruses. Possible reasons for the absolute requirement of the NS3 proteinase for NS4A in vitro are discussed.

* Author for correspondence. Fax +44 1707 332053. e-mail hilary.overton@roche.com

Received 9 May 1995; accepted 17 August 1995.


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