FIG. 3.

Rep-210-mediated inhibition of TYLCSV replication is not dependent on the oligomerization domain. (A) Southern blot of TNA extracts from wild-type N. benthamiana protoplasts cotransfected with pTOM6 together with the plasmid expressing the Rep mutant indicated above each lane; pGEM-P was used as a control. The blot was hybridized with a digoxigenin-labeled Rep-210 sense RNA probe. scDNA, super coiled DNA. (B) Quantitative analysis of TYLCSV replication in protoplasts cotransfected with pTOM6 together with the plasmid expressing the Rep mutant indicated on the left. Blots were hybridized with a ³²P-labeled DNA fragment corresponding to the Rep-54 coding sequence. Solid and open bars indicate the amount of ssDNA and scDNA respectively. The replication of TYLCSV in protoplasts cotransfected with pTOM6 and pGEM-P was taken as 100%. Each value is the average of two or three cotransfections of three independent experiments. Error bars indicate the standard error of the mean.