6533b7defe1ef96bd1275e7d

RESEARCH PRODUCT

A self-inducible heterologous protein expression system in Escherichia coli

Jean-marie HeydelGuillaume MarcionFabrice NeiersLoïc BriandYves ArturCarmen GarridoCarmen GarridoRenaud SeigneuricRenaud SeigneuricAlexandre Kriznik

subject

0106 biological sciences0301 basic medicineExpression systemslac operonHeterologousGene ExpressionmechanismLac repressorBiology[ SDV.MP.BAC ] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriologymedicine.disease_cause01 natural sciencesArticlelaw.inventionApplied microbiologylactose03 medical and health scienceslawlac repressor010608 biotechnologyt1r3 taste receptor[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN]Gene expressionmedicineEscherichia coliFood and NutritionInducerstationary-phaserecombinant geneinducerEscherichia coliMultidisciplinaryhsp70PromoterMolecular biology[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/BacteriologyRecombinant Proteins030104 developmental biologycloned genesBiochemistry[ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN]Alimentation et NutritionRecombinant DNA[SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN]bacteriophage-t7 rna-polymerase[SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology

description

AbstractEscherichia 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.

10.1038/srep33037https://hal-univ-bourgogne.archives-ouvertes.fr/hal-01392201