FIGURE 5.

Complementation of the ppi2 import deficiency in a transient expression system using protoplasts. Protoplasts derived from leaf cells of homozygous ppi2 plants were transformed with a plasmid encoding the N-terminal transit peptide of the small subunit of Rubisco fused to GFP (RbcS-nt:GFP) together with the empty vector (1) or together with different Toc159 constructs as indicated (2-7). Total protein extracts were prepared from protoplasts 12 h after transformation and subjected to Western blot analysis with anti-GFP and anti-Toc159 antibodies (for the upper panel, anti-atTOC159 serum was used, for the lower panel, affinity-purified anti-atTOC159 A-domain). ^*, proteolytic product of the mature form (26).