TY - JOUR
T1 - Controlled expression of recombinant proteins in Physcomitrella patens by a conditional heat-shock promoter
T2 - A tool for plant research and biotechnology
AU - Saidi, Younousse
AU - Finka, Andrija
AU - Chakhporanian, Mickhail
AU - Zrÿd, Jean Pierre
AU - Schaefer, Didier G.
AU - Goloubinoff, Pierre
PY - 2005/11
Y1 - 2005/11
N2 - 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.
AB - 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.
KW - β-glucuronidase
KW - Acetyl salicylic acid
KW - Actin cytoskeleton
KW - Benzyl alcohol
KW - GFP-talin
KW - Gmhsp17.3B promoter
KW - Inducible gene-expression system
UR - http://www.scopus.com/inward/record.url?scp=27644515982&partnerID=8YFLogxK
U2 - 10.1007/s11103-005-0889-z
DO - 10.1007/s11103-005-0889-z
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C2 - 16270224
AN - SCOPUS:27644515982
SN - 0167-4412
VL - 59
SP - 697
EP - 711
JO - Plant Molecular Biology
JF - Plant Molecular Biology
IS - 5
ER -