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. 2019 Jun 27;10:1472. doi: 10.3389/fmicb.2019.01472

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Copyright © 2019 Popa, Shi, Ruiz, Ferrer and Coca.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Ole18-CecA is susceptible to P. pastoris proteases. (A) Growth of recombinant strains carrying empty vector (EV), Olel8, Olel8-PAF102, Olel8-CecA. Aliquots of 5 μl of ten serial dilutions of log phase cultures (OD₆₀₀ = 1) were spotted on YPD agar media and allowed to grow for 2 days at 30°C. (B) Images of fluorescence and bright field microscopy of EV and Olel8-CecA cells after fluorescein diacetate (FDA) viability staining. (C) Immunoblot of OB fractions isolated from rice seeds producing Ole18-CecA^∗ or Ole18-PAF102^∗ fusion proteins, in the presence of total protein extracts (25 μg) from EV yeast cells, supplemented without (–) or with (+) protease inhibitors, incubated (I) or not (N) overnight at room temperature.