Controlled expression of recombinant proteins in Physcomitrella patens by a conditional heat-shock promoter: a tool for plant research and biotechnology.
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 beta-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 degrees C. In contrast, a short non-damaging heat-treatment at 38 degrees 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 degrees 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.
Saidi
Younousse
Y
Department of Plant Molecular Biology, Lausanne University, Lausanne CH-1015, Switzerland.
Finka
Andrija
A
Chakhporanian
Mickhail
M
Zrÿd
Jean-Pierre
JP
Schaefer
Didier G
DG
Goloubinoff
Pierre
P
D001241:Aspirin / Q000494:pharmacology; D019905:Benzyl Alcohol / Q000494:pharmacology; D001709:Biotechnology / Q000379:methods*; D019068:Bryopsida / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000235:genetics*; D006360:Heat-Shock Proteins / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D013696:Temperature; D013995:Time
D006360:Heat-Shock Proteins; D010940:Plant Proteins; D011994:Recombinant Proteins; D019905:Benzyl Alcohol; D001241:Aspirin