Figure 1

Design of TriTag-1 and TriTag-2, with alternatively spliced chloroplast-targeting PIMT2 and peroxisome-targeting TTL tags. (a, b) Splice diagrams of TriTag-1 (a) and TriTag-2 (b), showing non-targeting sequences (gray), chloroplast targeting sequences (light green), peroxisome targeting sequences (tan), and the enhanced GFP coding sequence used in transient expression experiments. (c, d) Design of TriTag-1 (c) and TriTag-2 (d) sequences. The ATG codon at the end corresponds to the first residue of the GFP open reading frame. Alternatively spliced targeting regions are highlighted. Donor and acceptor dimers are underlined. The light green DNA sequences derive from the PIMT2 5′ coding region [16] and include sequences required for chloroplast targeting (green). The light tan DNA sequences derive from the TTL 5′ coding region [21] and include sequences encoding the peroxisome targeting sequence (tan). (e, f) Final mRNAs species resulting from alternative splicing of TriTag-1 (e) and TriTag-2 (f).