Fig. 5.

In vitro packaging of ³²P-labelled rotavirus mRNA (transcribed from DLPs) into particles: Protection from RNase I digestion. 0.8% agarose gel, MOPS-Tris buffer 10 mM, pH 7.7. (A) Gel unfixed, stained with ethidium bromide; (B) Gel fixed, silver stained; (C) Gel fixed, dried and autoradiographed. Lane 1: DLPs; lane 2: native RV cores; lane 3: cores + RV mRNA + EGTA + packaging mixture; lane 4: as lane 3 + RNase I; lane 5: cores + mRNA + EGTA + packaging mixture + VP6; lane 6: as lane 5 + RNase I; lane 7: mRNA [the dsRNA seen in panels A and B originates from the DLPs which have been pelleted by ultracentrifugation after the in vitro transcription reaction and may have been partially damaged]; lane 8: as lane 7 + RNase I. Cores amount used: 60 ng/20 μl, or approximately 5 × 10⁸ particles.