Expression of neo transgenes in the
plastid genome. A, Immunoblot analysis to detect NPTII. Amount of total
soluble leaf protein (micrograms) loaded on the SDS-PAGE gel is
indicated above the lanes. A commercial NPTII dilution series was
loaded for reference. Lanes for plant lines are designated with
transforming plasmid; Wt, Wild-type tobacco sample. B, NPTII detected
by Coomassie Brilliant Blue R250 in SDS-PAGE gel. Twenty micrograms
total soluble protein was loaded per lane. Commercially available NPTII
(400 ng) was also loaded. Marked are the Rubisco large (LSU) and small
(SSU) subunits. C, The levels of neo mRNA in the
transplastomic leaves. Blots were probed for neo (top) and
cytoplasmic 25S rRNA as loading control (bottom). D, Levels of NPTII
and neo mRNA based on three to six experiments. Highest
value was taken as 100%. Plant line, plasmid name (for example pHK30)
and number and letter combination; + or −, SD is present or absent at
prokaryotic consensus position; wt or m, downstream sequence is wild
type or mutant.