Figure 6.

Transiently Expressed Sections of OEP80 and OEP24 Pull Down TOC Components.

(A) Diagram of constructs used (left). Chloroplasts from plants transiently expressing YFP-tagged constructs were solubilized with 1% (w/v) dodecyl-maltoside (input) and mixed with GFP affinity beads. Beads were washed, then bound proteins eluted. The inputs and eluates were subjected to immunoblotting with antibodies against GFP, Toc159, Tic40, and Toc75. The blot for GFP, Toc159, and Tic40 are from one membrane each; however, some regions were removed for clarity (indicated by dotted white and black lines). The asterisks indicate bands from the GFP affinity beads recognized by the Toc75 antibody. One of these comigrated with Toc75, but eluates 1, 2, and 3 showed a higher signal in this region. 80TP, transit peptide of OEP80; Plsp1TP, transit peptide of Plsp1; u, uncharacterized region between the transit peptide and the first POTRA. The three POTRA domains are labeled as 1, 2, and 3.

(B) Chloroplasts from uninfiltrated (UI) plants or plants transiently expressing T7-tagged OEP24 (24T7) or OEP40 (40T7) were solubilized with 1% (w/v) dodecyl-maltoside (input) and mixed with T7 affinity beads. The inputs and eluates were subjected to immunoblotting with antibodies against T7, Toc75, Toc159, and Tic40.