Large quantities of recombinant PR-5 proteins from the extracellular matrix of tobacco: Rapid production of microbial-recalcitrant proteins |
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Authors: | Albino Maggio Matilde Paino D'Urzo Laura R Abad Satomi Takeda Paul M Hasegawa Ray A Bressan |
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Institution: | (1) Center for Plant Environmental Stress Physiology, Purdue University, 1165 Horticulture Building, 47907-1165 West Lafayette, Indiana, USA |
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Abstract: | A procedure for the extraction of large quantities of PR-5 proteins that have been recalcitrant to microbial-based expression
systems is described. Targeting of the recombinant proteins to the extracellular matrix allowed efficient protein extraction
by a vacuum infiltration/centrifugation system. Approximately 1 kg of fresh leaves from transgenic tobacco plants overexpressing
either truncated osmotin (Liu et al., 1996) or A9 fromAtriplex nummularia L. (Casas et al., 1991) yielded between 3 and 5 mg of purified proteins that fully retained their antifungal activity. The
entire system of overexpression, extraction, and purification could be easily scaled up for the production of several grams
of protein. |
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Keywords: | PR-5 proteins transgenic tobacco extraction |
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