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Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology

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Abstract

The ability to express tightly controlled amounts of endogenous and recombinant proteins in plant cells is an essential tool for research and biotechnology. Here, the inducibility of the soybean heat-shock Gmhsp17.3B promoter was addressed in the moss Physcomitrella patens, using β-glucuronidase (GUS) and an F-actin marker (GFP-talin) as reporter proteins. In stably transformed moss lines, Gmhsp17.3B-driven GUS expression was extremely low at 25 °C. In contrast, a short non-damaging heat-treatment at 38 °C rapidly induced reporter expression over three orders of magnitude, enabling GUS accumulation and the labelling of F-actin cytoskeleton in all cell types and tissues. Induction levels were tightly proportional to the temperature and duration of the heat treatment, allowing fine-tuning of protein expression. Repeated heating/cooling cycles led to the massive GUS accumulation, up to 2.3% of the total soluble proteins. The anti-inflammatory drug acetyl salicylic acid (ASA) and the membrane-fluidiser benzyl alcohol (BA) also induced GUS expression at 25 °C, allowing the production of recombinant proteins without heat-treatment. The Gmhsp17.3B promoter thus provides a reliable versatile conditional promoter for the controlled expression of recombinant proteins in the moss P. patens.

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Abbreviations

ASA:

acetyl salicylic acid

BA:

benzyl alcohol

GFP:

green fluorescent protein

GT:

GFP-talin

GUS:

β-glucuronidase

Fv/Fm:

the maximal photochemical efficiency of PSII

MU:

4-methylumbelliferone

MUG:

4-methylumbelliferyl-β-D-glucuronide

PSII:

photosystem II

SA:

specific activity

sHSP:

small heat-shock protein

TSP:

total soluble proteins

Ubi:

ubiquitin

X-Gluc:

5-bromo-4-chloro-3-indolyl-β-D-glucuronide

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  1. Department of Plant Molecular Biology, Lausanne University, Lausanne, CH-1015, Switzerland

    Younousse Saidi, Andrija Finka, Mickhail Chakhporanian, Jean-Pierre Zrÿd, Didier G. Schaefer & Pierre Goloubinoff

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  1. Younousse Saidi
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  2. Andrija Finka
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  3. Mickhail Chakhporanian
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  4. Jean-Pierre Zrÿd
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  5. Didier G. Schaefer
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  6. Pierre Goloubinoff
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Correspondence to Pierre Goloubinoff.

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Saidi, Y., Finka, A., Chakhporanian, M. et al. Controlled Expression of Recombinant Proteins in Physcomitrella patens by a Conditional Heat-shock Promoter: a Tool for Plant Research and Biotechnology. Plant Mol Biol 59, 697–711 (2005). https://doi.org/10.1007/s11103-005-0889-z

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