A self-inducible heterologous protein expression system in Escherichia coli.
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-β-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.
Centre des Sciences du Goût et de l'Alimentation, INRA, Université de Bourgogne Franche-Comté, F-21000 Dijon, France.
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Briand
Université de Bourgogne Franche-Comté, Dijon, France.|INSERM, UMR 866, 7 blvd Jeanne d'Arc, 21000 Dijon, France.
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Marcion
UMR 7365 CNRS-Université de Lorraine IMoPA, 9 Avenue de la Forêt de Haye 54505 Vandoeuvre Les Nancy.
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Kriznik
Centre des Sciences du Goût et de l'Alimentation, INRA, Université de Bourgogne Franche-Comté, F-21000 Dijon, France.|Université de Bourgogne Franche-Comté, Dijon, France.
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Heydel
Centre des Sciences du Goût et de l'Alimentation, INRA, Université de Bourgogne Franche-Comté, F-21000 Dijon, France.|Université de Bourgogne Franche-Comté, Dijon, France.
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Artur
Université de Bourgogne Franche-Comté, Dijon, France.|INSERM, UMR 866, 7 blvd Jeanne d'Arc, 21000 Dijon, France.|Anticancer Center Georges François Leclerc, Dijon, France.
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Garrido
Université de Bourgogne Franche-Comté, Dijon, France.|INSERM, UMR 866, 7 blvd Jeanne d'Arc, 21000 Dijon, France.
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Seigneuric
Centre des Sciences du Goût et de l'Alimentation, INRA, Université de Bourgogne Franche-Comté, F-21000 Dijon, France.|Université de Bourgogne Franche-Comté, Dijon, France.
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Neiers
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D011994:Recombinant Proteins