FIG. 4.

Posttranslational membrane association of NS5B and the LVL mutant in vitro. IVTT reactions of pCMVNS5Bcon and -LVL were performed in the absence of microsomal membranes. After 1 h, translation was blocked by puromycin, followed by the addition of microsomal membranes (MM) to an aliquot of the reactions and incubation for one additional hour. Subsequently, membrane sedimentation analyses were performed as described in Materials and Methods. Supernatant (S) and pellet (P) fractions were analyzed by SDS-12% PAGE. Relative amounts of [³⁵S]methionine-labeled translation products, as quantified by phosphorimaging, are given at the bottom.