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. 2004 Aug;78(15):8254–8263. doi: 10.1128/JVI.78.15.8254-8263.2004

FIG. 6.

FIG. 6.

Mix and match in vitro replicase assays. The CNV replicase preparations represent enriched membrane fractions (lanes 1 to 5), affinity-purified proteins (lanes 6 to 10), and solubilized membrane fractions (lanes 11 to 17) obtained from yeast, except for lane 16, which contains products that were generated by CNV replicase obtained from N. benthamiana plants. The CNV replicase assay depicted in lanes 6 to 17 included exogenous RI/III(−) templates. The coexpressed p33, p92, and DI-72(+) RNA in yeast cells are shown on the top as plus signs, while yeast lacking one of these components is marked with minus signs. The boxed lanes represent the mixed preparations as follows. Preparations containing either p33 or p92 (coexpressed with DI-72 RNA) were obtained from separate yeast lines, which were mixed in 1:1 ratio prior to the CNV replicase reaction (replicase products in lanes 3, 6, 14, and 15 are from mixing preparations corresponding to yeast lines shown in lanes 4 and 5; 7 and 8; and 12 and 13, respectively). Note that the CNV replicase preparation in lane 15 was obtained by mixing p92 and p33 preparations in a 1:4 ratio. The CNV replicase products generated on endogenous DI-72 or exogenous RI/III(−) templates are marked. The lane marked Nb represents the replicase product obtained with the plant-derived CNV replicase preparation.