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PMID: 16270224 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Controlled expression of recombinant proteins in Physcomitrella patens by a conditional heat-shock promoter: a tool for plant research and biotechnology.

Plant molecular biology ·Vol. 59 ·No. 5 ·2005-11-00 ·Pages 697-711

Saidi Y, Finka A, Chakhporanian M, Zrÿd JP, Schaefer DG, Goloubinoff P

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 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.

MeSH Terms
Aspirin/pharmacology Benzyl Alcohol/pharmacology Biotechnology/methods Bryopsida/drug effects,genetics,metabolism Dose-Response Relationship, Drug Enzyme Induction Gene Expression Regulation, Enzymologic/genetics Gene Expression Regulation, Plant/drug effects,genetics Heat-Shock Proteins/genetics Plant Proteins/genetics,metabolism Promoter Regions, Genetic/genetics Recombinant Proteins/genetics,metabolism Temperature Time
Chemicals
Heat-Shock Proteins Plant Proteins Recombinant Proteins Benzyl Alcohol Aspirin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Saidi Younousse
Department of Plant Molecular Biology, Lausanne University, Lausanne CH-1015, Switzerland.
Finka Andrija
Chakhporanian Mickhail
Zrÿd Jean-Pierre
Schaefer Didier G
Goloubinoff Pierre
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Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
DOI
10.1007/s11103-005-0889-z
Published
2005-11-00
Pages
697-711
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
External Links
PubMed source DOI: 10.1007/s11103-005-0889-z View journal details
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