Figure 2. Molecular characterization of acn2.

(A) The acn2 locus. DNA and total RNA were isolated from Col-0 and acn2 mutants. All exons of the acn2 locus were sequenced once, and the mutation was confirmed by re-sequencing of exon 13 and the corresponding region of the cDNA. Exons and introns are represented by open and closed bars respectively. (B) An ethidium-bromide-stained agarose gel showing the expression of CTS as determined by RT–PCR. rbcS represents the small subunit of ribulose bisphosphate carboxylase. (C–E) Analysis of CTS protein in PGS 0.7 seedlings germinated on standard agar medium plates containing 20 mM sucrose using either C-terminal- (C) or N-terminal- (D) specific antibodies. Lane 1, Ler; Lane 2, cts-1; Lane 3, Col-0; Lane 4, acn2. Protein (20 μg) was loaded into each well, and equal loading was verified by staining with Ponceau S (E).