Figure 1
From: Plant-made Salmonella bacteriocins salmocins for control of Salmonella pathovars
Expression of salmocins in plants. Transient expression in N. benthamiana upon vacuum infiltration (a) or syringe infiltration (b) with agrobacteria carrying TMV or TMV and PVX vectors. Coomassie-stained SDS protein gels loaded with samples (a,b) corresponding to 3 mg FW plant material, (a) crude extracts prepared with 2× Laemmli buffer or (b) TSP extracts prepared with 50 mM HEPES pH 7.0, 10 mM K acetate, 5 mM Mg acetate, 10% (v/v) glycerol, 0.05% (v/v) Tween 20®, 300 mM NaCl. (c) Inducible expression of salmocin SalE1b in stable transgenic Nicotiana benthamiana plants. Loading with crude extracts corresponds to 3 mg FW extracted with 2× Laemmli buffer from (lanes 1, 3, 5, 7) non-induced plant material or (lanes 2, 4, 6, 8) plant material 4 dp induction with ethanol. (lanes 1, 2) N. benthamiana WT plant, (lanes 3, 4), (lanes 5, 6), (lanes 7, 8) different transgenic plant candidates for single copy T-DNA insertion of T0 generation (#4, 12, 37 for SalE1b). (d) Transient expression in Spinacia oleracea cv. Frühes Riesenblatt upon syringe infiltration with agrobacteria carrying TMV or TMV and PVX vectors. Loading of TSP extracts corresponds to 3 mg FW plant material extracted with 5 vol. 150 mM NaCl. Plant material was harvested (a) 5 dpi (days post infiltration) for SalE1b, 6 dpi for SalE3, SalE7 and SalE1a or 7 dpi for SalE2 or (b) 4 dpi for SalE1b, 5 dpi for SalE3, SalE7 and SalE1a and 6 dpi for SalE2 or (d) 8 dpi for SalE2, SalE3, SalE7, SalE1a and SalE1b. (a,b,d) Analysed extracts were prepared from plant material expressing SalE2 (lane 1), SalE3 (lane 2), SalE7 (lane 3), SalE1a (lane 4) and SalE1b (lane 5) or from (WT) non-transfected leaf tissue. SalE2 and SalE7 were co-expressed with their respective immunity proteins. Asterisks mark recombinant proteins.
