ArticlePloS one2016
Biochemical Preparation of Cell Extract for Cell-Free Protein Synthesis without Physical Disruption.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Article
- An Optimized and Systematically Integrated Expression, Extraction, and Purification Strategy Enabling Efficient Production of TCR-Like Antibodies in Escherichia coli.Current microbiology · 2026Article
- A Pichia pastoris Cell-Free Protein Synthesis Platform.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Cell-free expression system: a promising platform for bacteriophage production and engineering.Microbial cell factories · 2025Review
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- The extracts of osteoblast developed from adipose-derived stem cell and its role in osteogenesis.Journal of orthopaedic surgery and research · 2024Article
- Multimolecular Competition Effect as a Modulator of Protein Localization and Biochemical Networks in Cell-Size Space.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- 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
- Complex Dependence ofACS synthetic biology · 2023Article
- Established and Emerging Methods for Protecting Linear DNA in Cell-Free Expression Systems.Methods and protocols · 2023Review
- On-demand biomanufacturing through synthetic biology approach.Materials today. Bio · 2023Article
- Key reaction components affect the kinetics and performance robustness of cell-free protein synthesis reactions.Computational and structural biotechnology journal · 2022Article
- Activation of a dilutedSTAR protocols · 2021Article
- Evaluation of an E. coli Cell Extract Prepared by Lysozyme-Assisted Sonication via Gene Expression, Phage Assembly and Proteomics.Chembiochem : a European journal of chemical biology · 2021Article
- System concentration shift as a regulator of transcription-translation system within liposomes.iScience · 2021Article
- A Relationship between NTP and Cell Extract Concentration for Cell-Free Protein Expression.Life (Basel, Switzerland) · 2021Article
- Methodologies for preparation of prokaryotic extracts for cell-free expression systems.Synthetic and systems biotechnology · 2020Review
- Review
- A PEGDA/DNA Hybrid Hydrogel for Cell-Free Protein Synthesis.Frontiers in chemistry · 2020Article
- Article
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cell-free protein synthesis (CFPS) is a powerful tool for the preparation of toxic proteins, directed protein evolution, and bottom-up synthetic biology. The transcription-translation machinery for CFPS is provided by cell extracts, which usually contain 20-30 mg/mL of proteins. In general, these cell extracts are prepared by physical disruption; however, this requires technical experience and special machinery. Here, we report a method to prepare cell extracts for CFPS using a biochemical method, which disrupts cells through the combination of lysozyme treatment, osmotic shock, and freeze-thaw cycles. The resulting cell extracts showed similar features to those obtained by physical disruption, and was able to synthesize active green fluorescent proteins in the presence of appropriate chemicals to a concentration of 20 μM (0.5 mg/mL).
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