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. 2021 Jul 14;12:703020. doi: 10.3389/fpls.2021.703020

Figure 1.

Figure 1

Expression cassette and confirmation of acid-alpha glucosidase (GAA) production in the transformed Arabidopsis alg3 cell culture. (A) Schematic representation of pFK1-BAR-GAA. Cauliflower mosaic virus (CaMV) 35Sp: CaMV 35S promoter; AtADH 5’-UTR: 5'-untranslated region of Arabidopsis alcohol dehydrogenase gene which acts as an enhancer; GAA: cDNA of human GAA; HSP: Arabidopsis heat shock protein terminator; and LB and RB: left and right borders of T-DNA, respectively. (B,C) CBB staining and Western blot analysis of untransformed (wt) and transformed (GAA) 7-day-old Arabidopsis alg3 cell culture to confirm GAA production intra (int)- and extracellularly (ext). Marks a, b, and c indicate the 110, 95, and 76 kDa GAAs, respectively. C+ is human embryonic kidney/HEK293-derived recombinant human GAA used as a positive control. (D,E) GAA-specific activities in crude intracellular and extracellular proteins of untransformed (wt) and transformed (GAA) Arabidopsis alg3 cell culture.