FIG. 3.

Synthesis, transport, and release of structural proteins following transfection of mutant RNAs. BHK cells were transfected with in vitro-transcribed mutant or parental BRM34 RNAs by electroporation. At 40 h postinfection, the infected cells were pulse-labeled for 80 min with [³⁵S]methionine and chased with medium containing unlabeled methionine for 10 min or 4 h. The chase medium was harvested, and the labeled cells were lysed with lysis buffer. The lysates were immunoprecipitated with human anti-RV serum and analyzed by SDS-PAGE on 10% gels under reducing conditions. Virus particles in the chase medium were precipitated with PEG, and pelleted virus particles were resuspended in Triton-TNE buffer. The suspension was immunoprecipitated with human anti-RV serum and analyzed by SDS-PAGE (10% gel) and subsequent autoradiography. Positions of migration of RV structural proteins E1, E2, and C are shown.