A self-inducible heterologous protein expression system in Escherichia coli

Loïc Briand (1) , Guillaume Marcion (2) , Alexandre Kriznik (3) , Jean-Marie Heydel (1) , Yves Artur (1) , Carmen Garrido (4, 2) , Renaud Seigneuric (2) , Fabrice Neiers (1)

1 CSGA - Centre des Sciences du Goût et de l'Alimentation [Dijon]
2 Protéines de choc thermique : mort cellulaire, différenciation cellulaire et propriétés tumorigéniques (U866, Cancer, équipe 3)
3 IMoPA - Ingénierie Moléculaire et Physiopathologie Articulaire
4 UNICANCER/CRLCC-CGFL - Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon]

Escherichia coli is an important experimental, medical and industrial cell factory for recombinant protein production. The inducible lac promoter is one of the most commonly used promoters for heterologous protein expression in E. coli. Isopropyl-beta-D-thiogalactoside (IPTG) is currently the most efficient molecular inducer for regulating this promoter's transcriptional activity. However, limitations have been observed in large-scale and microplate production, including toxicity, cost and culture monitoring. Here, we report the novel SILEX (Self-InducibLe Expression) system, which is a convenient, cost-effective alternative that does not require cell density monitoring or IPTG induction. We demonstrate the broad utility of the presented self-inducible method for a panel of diverse proteins produced in large amounts. The SILEX system is compatible with all classical culture media and growth temperatures and allows protein expression modulation. Importantly, the SILEX system is proven to be efficient for protein expression screening on a microplate scale.

Mots clés: stationary-phase; t1r3 taste receptor; recombinant gene; Expression systems; bacteriophage-t7 rna-polymerase; Applied microbiology; lac repressor; cloned genes; inducer; lactose; hsp70; mechanism

Domaines: Bactériologie; Génomique, Transcriptomique et Protéomique [q-bio.GN]

Loïc Briand, Guillaume Marcion, Alexandre Kriznik, Jean-Marie Heydel, Yves Artur, et al.. A self-inducible heterologous protein expression system in Escherichia coli. Scientific Reports, 2016, 6, pp.33037. ⟨10.1038/srep33037⟩. ⟨hal-01392201⟩