ArticleMicrobial biotechnology2015
A chromosomally encoded T7 RNA polymerase-dependent gene expression system for Corynebacterium glutamicum: construction and comparative evaluation at the single-cell level.
Article in Microbial biotechnology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed, 91 citations in OpenAlex.
- Molecular Basis of pH-Dependent Activity in T7 RNA Polymerase Revealed by Constant pH Replica Exchange Molecular Dynamics.ACS physical chemistry Au · 2026Article
- Rapid Identification of Superior Endogenous Signal Peptides for Heterologous Protein Secretion by Corynebacterium glutamicum Through Modular Cloning and Automation.Microbial biotechnology · 2026Article
- Engineering T7 RNA polymerase-cascaded systems controlled by nisin and theophylline for protein overexpression and targeted gene mutagenesis inSynthetic and systems biotechnology · 2025Article
- Reduced Genetic Heterogeneity for Stable Bioproduction by Harnessing the Bias and Mechanism of Mutation.Microbial biotechnology · 2025Article
- Improving polyketide biosynthesis by rescuing the translation of truncated mRNAs into functional polyketide synthase subunits.Nature communications · 2025Article
- A cross-species inducible system for enhanced protein expression and multiplexed metabolic pathway fine-tuning in bacteria.Nucleic acids research · 2025Article
- OptoLacI: optogenetically engineered lactose operon repressor LacI responsive to light instead of IPTG.Nucleic acids research · 2024Article
- A type III polyketide synthase cluster in the phylum Planctomycetota is involved in alkylresorcinol biosynthesis.Applied microbiology and biotechnology · 2024Article
- Production of Double-Stranded RNA Using the Prokaryotic Promoter-Mediated Bidirectional Transcription.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum.Microbial cell factories · 2023Article
- Growth-rate dependency of ribosome abundance and translation elongation rate in Corynebacterium glutamicum differs from that in Escherichia coli.Nature communications · 2023Article
- Screening and identification of potential MERS-CoV papain-like protease (PLpro) inhibitors; Steady-state kinetic and Molecular dynamic studies.Saudi pharmaceutical journal : SPJ : the official publication of the Saudi Pharmaceutical Society · 2023Article
- High-level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source.Microbial biotechnology · 2022Article
- Physiological, Biochemical, and Structural Bioinformatic Analysis of the Multiple Inositol Dehydrogenases from Corynebacterium glutamicum.Microbiology spectrum · 2022Article
- A manually curated compendium of expression profiles for the microbial cell factory Corynebacterium glutamicum.Scientific data · 2022Article
- Biosynthesis of eriodictyol from tyrosine by Corynebacterium glutamicum.Microbial cell factories · 2022Article
- Customized chitooligosaccharide production-controlling their lengthFrontiers in bioengineering and biotechnology · 2022Article
- Highly tunable TetR-dependent target gene expression in the acetic acid bacterium Gluconobacter oxydans.Applied microbiology and biotechnology · 2021Article
- Efficient production of long double-stranded RNAs applicable to agricultural pest control by Corynebacterium glutamicum equipped with coliphage T7-expression system.Applied microbiology and biotechnology · 2021Article
- A Green Light-Regulated T7 RNA Polymerase Gene Expression System for Cyanobacteria.Marine biotechnology (New York, N.Y.) · 2021Article
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4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Corynebacterium glutamicum has become a favourite model organism in white biotechnology. Nevertheless, only few systems for the regulatable (over)expression of homologous and heterologous genes are currently available, all of which are based on the endogenous RNA polymerase. In this study, we developed an isopropyl-β-D-1-thiogalactopyranosid (IPTG)-inducible T7 expression system in the prophage-free strain C. glutamicum MB001. For this purpose, part of the DE3 region of Escherichia coli BL21(DE3) including the T7 RNA polymerase gene 1 under control of the lacUV5 promoter was integrated into the chromosome, resulting in strain MB001(DE3). Furthermore, the expression vector pMKEx2 was constructed allowing cloning of target genes under the control of the T7lac promoter. The properties of the system were evaluated using eyfp as heterologous target gene. Without induction, the system was tightly repressed, resulting in a very low specific eYFP fluorescence (= fluorescence per cell density). After maximal induction with IPTG, the specific fluorescence increased 450-fold compared with the uninduced state and was about 3.5 times higher than in control strains expressing eyfp under control of the IPTG-induced tac promoter with the endogenous RNA polymerase. Flow cytometry revealed that T7-based eyfp expression resulted in a highly uniform population, with 99% of all cells showing high fluorescence. Besides eyfp, the functionality of the corynebacterial T7 expression system was also successfully demonstrated by overexpression of the C. glutamicum pyk gene for pyruvate kinase, which led to an increase of the specific activity from 2.6 to 135 U mg(-1). It thus presents an efficient new tool for protein overproduction, metabolic engineering and synthetic biology approaches with C. glutamicum.
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