(A) Structure of the BMV RNA genome and plasmid DNA
cassettes used to express wild-type BMV RNA2 and CP.
GAL1, Rz, and A+ denote the GAL1
promoter, a self-cleaving hepatitis δ virus ribozyme (18), and the
yeast ADH1 polyadenylation signal, respectively (14).
(B) BMV RNA and protein accumulation in yeast. Equal
amounts of total RNA from yeast expressing the indicated BMV components
were analyzed by Northern blotting with RNA2 (Top) and
CP (Second blot) probes. RNA from plant-derived BMV
virions (BMVP) was used as a control. Equal amounts of
total protein from yeast expressing the indicated components were
analyzed by Western blotting with anti-2a (Third blot)
and anti-CP (Bottom) antisera. Controls (lane 5) were
baculovirus-expressed 2a protein (19) (Third blot, lane
5) and BMVP CP (Bottom, lane 5).