Fig 8.

Effect of the N- and C-terminal mutants of CP on replication and packaging. (A) Northern blot analysis of FHV RNA replication in plant cells. The agrotransformant of F1 was mixed with either wt F2 or the indicated mutants and infiltrated into N. benthamiana. At 5 dpi, the total RNAs were extracted and subjected to Northern blot hybridization with riboprobes specific for F1 and F2. The FHV RNA accumulation levels shown below the Northern blot were normalized against the wt as 100%. (B) Virion yield. FHV virions were purified from the infiltrated leaves, and the yield was quantitated and normalized against the wt as 100%. The data shown represent averages of three independent virion preparations. (C) Analyses of RNAs extracted from purified FHV virions. At 5 dpi, virions were purified from the infiltrated leaves, and encapsidated RNA was isolated. Virion RNA was subjected to 1% agarose electrophoresis and visualized by staining with ethidium bromide. An asterisk indicates a heterogeneous population RNA of cellular origin. (D) The RNA profile shown in panel C was subjected to Northern blot hybridization with a mixture of riboprobes complementary to full-length F1 and F2. The positions of F1, F2, and sgF3 are shown to the right. In panel A, rRNA indicates a loading control. Encapsidated FHV progeny was quantitated as described above.