Fig. 1.
Plasmid constructs used for the expression of Art v 1 in plants. Plasmids derived from pBI121: the entire ORF of Art v 1 including the endogenous signal peptide was placed between the restriction sites BamHI and SstI under the control of the 35S cauliflower mosaic promoter (P35S). Alternatively, the Art v 1 signal peptide was fused to the N-terminus of GFP (pBI121_SpArtv1::GFP), also under control of P35S. RB right border, LB left border, T NOS terminator of nopaline synthase, P NOS promoter of nopaline synthase, NPT II kanamycin resistance gene. Plasmids derived from pBINPLUS and pIMPACT series: coding sequence of Art v 1 without the signal peptide was cloned between NcoI and BglII sites in pIMPACT 1.3 resulting in an ER-localised protein. Expression is under control of the chrysanthemum promoter rbcS1 (PRbcS1). A signal peptide from sea anemona (SPsea) directs the protein of interest to the secretory pathway and the KDEL peptide retains the protein in the ER. All proteins are expressed with a C-terminal c-myc tag and 6xHis tag