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. 2014 Apr 15;15(7):650–663. doi: 10.1111/mpp.12122

Figure 3.

figure

Characterization of the recombinant ToxA proteins. (A) Sodium dodecylsulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) gel showing ToxA and the ToxA‐N102A mutant proteins expressed in Pichia pastoris. Each lane was loaded with 20 μL of cell‐free culture supernatant. The control sample was from a ‘mock’ strain transformed with the expression vector only. Numbers on the left indicate the molecular masses (in kDa) of protein markers. (B) Matrix‐assisted laser desorption ionization‐time of flight/time of flight mass spectrometry (MALDI‐TOF/TOF MS) data showing the 11‐amino‐acid fragments containing the wild‐type sequence (top, peak = 1274.71) and the alanine substitution (highlighted in red) in the mutant protein (bottom, peak = 1231.75). (C) Plant assays showing the activity of the recombinant ToxA proteins. ToxA‐sensitive wheat (Grandin) plants were infiltrated with ToxA at the estimated final concentrations (ng/μL) indicated on the right. Control plants were treated with the protein sample of the ‘mock’ strain shown in (A) (lane 3). Marks delimit the infiltrated area. Photographs were taken at 72 h post‐infiltration.