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Processing of the NS3-4A junction via a trans mechanism. Construct iC-54, with the S139A protease active-site mutation, was translated for 60 min. A 20-μl aliquot of the lysate was incubated with (+) or without (−) the purified sc-NS4A21-32-GSGS-NS33-181 protein (final concentration, 250 nM) for 60 min at 30°C. The products were separated by SDS-8% polyacrylamide gels and analyzed by autoradiography.
