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. 2018 Dec 6;17(6):1069–1080. doi: 10.1111/pbi.13038

Figure 3.

Figure 3

Production of large amounts of a Penicillium digitatum unknown protein in Nicotiana benthamiana leaves. (a) Schematic representation of the constructs generated to accumulate the protein of unknown function (UP, PDIG_23520) in the apoplast, ER, or vacuole of plant cells. The three constructs include the signal peptide of the tobacco AP24 protein at the N‐terminus. A KDEL signal was added at the C‐terminus in the construct designed for ER‐retention, and the vacuolar signal (VS) was fused at the C‐end for vacuolar accumulation. (b) Appearance of N. benthamiana leaves agroinfiltrated with the indicated constructs after 7 days. (c) Analysis of UP accumulation in agroinfiltrated leaves. Proteins from extracellular fluids (ECFs, left panel) or acid extraction (right panel) were separated by tricine‐SDS‐PAGE and Coomassie‐blue stained. Purified UP from P. digitatum cultures was run in parallel as a control. Molecular weight markers are shown in kDa on the left.