FIG. 2.

Self-cleavage at the NS3-4A junction is insensitive to dilution. (A) One microgram of RNA encoding T-54 or C-54 was used to prime the in vitro translation reaction. Proteins were analyzed after 15, 30, and 60 min by electrophoresis in an SDS-8% polyacrylamide gel and autoradiography. The number above each lane refers to the reaction time (in minutes) for the sample. (B) Analysis of T-34 and C-34 constructs, processed in the same manner as for panel A. (C) Translation reactions programmed with T-54 and C-54 transcripts were carried out as described in Materials and Methods and were terminated after 15 min. The proteins were diluted 1-, 3-, 10- and 50-fold with buffer A and then incubated at 30°C for another 60 min before being precipitated with acetone and analyzed by electrophoresis. The numbers above the lanes indicate the dilution factors. Pre, starting translation product prior to dilution and the second incubation.