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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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Bibliographic Details
Title: Controlled expression of recombinant proteins in Physcomitrella patens by a conditional heat-shock promoter: a tool for plant research and biotechnology.
Authors: Saidi, Y., Finka, A., Chakhporanian, M., Zrÿd, J.P., Schaefer, D.G., Goloubinoff, P.
Publication Year: 2025
Collection: Université de Lausanne (UNIL): Serval - Serveur académique lausannois
Subject Terms: Aspirin/pharmacology, Benzyl Alcohol/pharmacology, Biotechnology/methods, Bryopsida/drug effects, Bryopsida/genetics, Dose-Response Relationship, Drug, Enzyme Induction, Gene Expression Regulation, Enzymologic/genetics, Plant/drug effects, Plant/genetics, Heat-Shock Proteins/genetics, Plant Proteins/genetics, Plant Proteins/metabolism, Promoter Regions, Genetic/genetics, Recombinant Proteins/genetics, Recombinant Proteins/metabolism, Temperature, Time
Description: 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.
Document Type: article in journal/newspaper
File Description: application/pdf
Language: English
ISSN: 0167-4412
Relation: Plant Molecular Biology; https://iris.unil.ch/handle/iris/152911; serval:BIB_9E1288D32DF8; 000233044100001
DOI: 10.1007/s11103-005-0889-z
Availability: https://iris.unil.ch/handle/iris/152911
https://doi.org/10.1007/s11103-005-0889-z
Accession Number: edsbas.29BF3
Database: BASE
Description
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.
ISSN:01674412
DOI:10.1007/s11103-005-0889-z