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

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

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)697-711
Number of pages15
JournalPlant Molecular Biology
Volume59
Issue number5
DOIs
StatePublished - Nov 2005
Externally publishedYes

Keywords

  • Acetyl salicylic acid
  • Actin cytoskeleton
  • Benzyl alcohol
  • GFP-talin
  • Gmhsp17.3B promoter
  • Inducible gene-expression system
  • β-glucuronidase

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