ArticleScientific reports2017
Cell-free protein synthesis from non-growing, stressed Escherichia coli.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- Characterizing Cell-Free Transcription and Translation Dynamics with Nucleic Acid-Based Assays.ACS synthetic biology · 2026Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Designing of an extract production protocol for industrial application of cell-free protein synthesis technology: Building from a current best practice to a quality by design approach.Engineering biology · 2023Review
- Establishing a Cell-Free Transcription-Translation Platform forACS biomaterials science & engineering · 2023Article
- What remains from living cells in bacterial lysate-based cell-free systems.Computational and structural biotechnology journal · 2023Review
- Systems biology-based analysis of cell-free systems.Current opinion in biotechnology · 2022Review
- Ribosome purification fromBiotechnology notes (Amsterdam, Netherlands) · 2022Article
- Key reaction components affect the kinetics and performance robustness of cell-free protein synthesis reactions.Computational and structural biotechnology journal · 2022Article
- Cell-Free PURE System: Evolution and Achievements.Biodesign research · 2022Review
- Preparation and Screening of Cell-Free Extract from Nongrowing Escherichia coli A19 Cells.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Cell-Free Protein Synthesis by Diversifying Bacterial Transcription Machinery.Biotech (Basel (Switzerland)) · 2021Article
- Development and comparison of cell-free protein synthesis systems derived from typical bacterial chassis.Bioresources and bioprocessing · 2021Article
- Methodologies for preparation of prokaryotic extracts for cell-free expression systems.Synthetic and systems biotechnology · 2020Review
- Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology.Frontiers in bioengineering and biotechnology · 2020Review
- Screening and Identification of Novel cGAS Homologues Using a Combination of in Vitro and In Vivo Protein Synthesis.International journal of molecular sciences · 2019Article
- Methods to reduce variability inSynthetic and systems biotechnology · 2019Article
- Cell-free Protein Expression Using the Rapidly Growing Bacterium Vibrio natriegens.Journal of visualized experiments : JoVE · 2019Article
- Deconstructing Cell-Free Extract Preparation for in Vitro Activation of Transcriptional Genetic Circuitry.ACS synthetic biology · 2019Article
- Cell-Free Protein Synthesis From Fast-GrowingFrontiers in microbiology · 2018Article
- Elimination of Ribosome Inactivating Factors Improves the Efficiency ofFrontiers in microbiology · 2018Article
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cell-free protein synthesis is a versatile protein production system. Performance of the protein synthesis depends on highly active cytoplasmic extracts. Extracts from E. coli are believed to work best; they are routinely obtained from exponential growing cells, aiming to capture the most active translation system. Here, we report an active cell-free protein synthesis system derived from cells harvested at non-growth, stressed conditions. We found a downshift of ribosomes and proteins. However, a characterization revealed that the stoichiometry of ribosomes and key translation factors was conserved, pointing to a fully intact translation system. This was emphasized by synthesis rates, which were comparable to those of systems obtained from fast-growing cells. Our approach is less laborious than traditional extract preparation methods and multiplies the yield of extract per cultivation. This simplified growth protocol has the potential to attract new entrants to cell-free protein synthesis and to broaden the pool of applications. In this respect, a translation system originating from heat stressed, non-growing E. coli enabled an extension of endogenous transcription units. This was demonstrated by the sigma factor depending activation of parallel transcription. Our cell-free expression platform adds to the existing versatility of cell-free translation systems and presents a tool for cell-free biology.
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