Accumulation of the p25 CP in each transgenic
line was analyzed by Western blot. Leaf tissue was
ground with extraction buffer consisting of 0.1 M
Tris-HCl pH 6.8 and 1 mM phenyl-methyl-sulfonyl-
fluoride (PMSF). Protein extracts were electrophoresed
in 15% SDS-polyacrilamide gels, and electroblotted
to Immobilon-PVDF membranes (Millipore).
Proteins were probed with the monoclonal antibodies
3DF1 + 3CA5 to the CTV p25 CP protein (Cambra
et al. 1990; Vela et al. 1986) as the primary antibodies,
and goat anti-mouse IgG conjugated with alkaline
phosphatase as the secondary antibody