ArticleAdvanced structural and chemical imaging2018
Protein structural biology using cell-free platform from wheat germ.
Article in Advanced structural and chemical imaging, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Eukaryotic cell-free protein synthesis: Chassis diversification, system engineering, and emerging applications.Synthetic and systems biotechnology · 2026Review
- Half a Century of Progress: The Evolution of Wheat Germ-Based In Vitro Translation into a Versatile Protein Production Method.International journal of molecular sciences · 2025Review
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Unlocking the distinctive enzymatic functions of the early plant biomass deconstructive genes in a brown rot fungus by cell-free protein expression.Applied and environmental microbiology · 2024Article
- Opportunities to accelerate extracellular vesicle research with cell-free synthetic biology.Journal of extracellular biology · 2023Article
- The Use of Cell-free Protein Synthesis to Push the Boundaries of Synthetic Biology.Biotechnology and bioprocess engineering : BBE · 2023Review
- Multiple Gene Expression in Cell-Free Protein Synthesis Systems for Reconstructing Bacteriophages and Metabolic Pathways.Microorganisms · 2022Review
- Cell-free protein crystallization for nanocrystal structure determination.Scientific reports · 2022Article
- Cryo-EM structure of the diapause chaperone artemin.Frontiers in molecular biosciences · 2022Article
- Bacteriophage-encoded lethal membrane disruptors: Advances in understanding and potential applications.Frontiers in microbiology · 2022Review
- Tunable Heteroassembly of a Plant Pseudoenzyme-Enzyme Complex.ACS chemical biology · 2021Article
- Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology.Frontiers in molecular biosciences · 2021Review
- Serodiagnostic antigens of Clonorchis sinensis identified and evaluated by high-throughput proteogenomics.PLoS neglected tropical diseases · 2020Article
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Authors and funding
11 authors.
Funding
Abstract
One of the biggest bottlenecks for structural analysis of proteins remains the creation of high-yield and high-purity samples of the target protein. Cell-free protein synthesis technologies are powerful and customizable platforms for obtaining functional proteins of interest in short timeframes, while avoiding potential toxicity issues and permitting high-throughput screening. These methods have benefited many areas of genomic and proteomics research, therapeutics, vaccine development and protein chip constructions. In this work, we demonstrate a versatile and multiscale eukaryotic wheat germ cell-free protein expression pipeline to generate functional proteins of different sizes from multiple host organism and DNA source origins. We also report on a robust purification procedure, which can produce highly pure (> 98%) proteins with no specialized equipment required and minimal time invested. This pipeline successfully produced and analyzed proteins in all three major geometry formats used for structural biology including single particle analysis with electron microscopy, and both two-dimensional and three-dimensional protein crystallography. The flexibility of the wheat germ system in combination with the multiscale pipeline described here provides a new workflow for rapid production and purification of samples that may not be amenable to other recombinant approaches for structural characterization.
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Registered trials
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