Effect of ORN on RNA packaging in cell-free assembly assay.
35S-labelled in vitro assembled BTV complexes were fractionated in a continuous sucrose gradient . (Upper panel): Fractions #5, #6 and #7 from cell-free assembly (CFA) reactions in the absence (+control, lanes 1 to 3) or presence of 20 pmol S10.2 ORN (lanes 4 to 6) alongside with fraction #6 in the presence of S10.4 ORN (lane 7) S10.5 (lane 8) and Scr ORN (lane 9) were analysed on 1% denaturing agarose gel. Packaged RNAs were determined by densitometry. Lower panel represents the mean values (%) of total packaged ssRNAs in the presence of ORNs calculated relative to the control reaction (without ORNs) set at 100% (n = 3). The synthesized 35S-labelled BTV subcore and transcription complex protein profile are shown in S8 Fig.