Fig. 5.
BaMV replication in protoplasts of N. benthamiana. (A) Accumulation of viral coat protein. Protoplasts (3 × 105) were inoculated with 2 μg pCBG or its derivative as indicated and 6 μg carrier DNA and incubated at room temperature for 48 h. The protein extracts (5 μg each) of the protoplasts were then analyzed by Western blot using anti-coat protein antibodies and chemiluminescence reagent. The relative amounts of the viral coat protein were estimated according to the luminescent signals by referring WT as 100%. (B) Accumulation of positive strand viral RNAs. Total RNA (0.5 μg) from protoplasts infected with pCBG or the indicated derivative was analyzed by Northern blot using a 0.6 kb RNA probe complementary to the 3′-terminal region of the genomic RNA. The viral genome (gRNA) and two subgenomic RNA (sg1 and sg2) are indicated. The relative amounts of the genomic RNA were estimated according to the respective pixels by considering WT as 100%.