ArticleJournal of structural and functional genomics2004
High-throughput, genome-scale protein production method based on the wheat germ cell-free expression system.
Article in Journal of structural and functional genomics, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology.Frontiers in molecular biosciences · 2021Review
- KLR-70: A Novel Cationic Inhibitor of the Bacterial Hsp70 Chaperone.Biochemistry · 2020Article
- Study roadmap for high-throughput development of easy to use and affordable biomarkers as diagnostics for tropical diseases: a focus on malaria and schistosomiasis.Infectious diseases of poverty · 2017Article
- Studying macromolecular complex stoichiometries by peptide-based mass spectrometry.Proteomics · 2015Review
- Artificial environments for the co-translational stabilization of cell-free expressed proteins.PloS one · 2013Article
- Using translational enhancers to increase transgene expression in Drosophila.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- FLEXIQuant: a novel tool for the absolute quantification of proteins, and the simultaneous identification and quantification of potentially modified peptides.Journal of proteome research · 2009Article
- A robust two-step PCR method of template DNA production for high-throughput cell-free protein synthesis.Journal of structural and functional genomics · 2007Article
- Pilot studies on the parallel production of soluble mouse proteins in a bacterial expression system.Journal of structural and functional genomics · 2005Article
Corrections and comments
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Current cell-free protein expression systems are capable of synthesizing proteins with high speed and accuracy; however, the yields are low due to their instability over time. Escherichia coli based systems are not always sufficient for expression of eukaryotic proteins. This report reviews a high-throughput protein production method based on the cell-free system prepared from eukaryote, wheat embryos. We first demonstrate a method for preparation of this extract that exhibited a high degree of stability and activity. To maximize translation yield and throughput, we address and resolve the following issues: (1) optimization of the ORF flanking regions; (2) PCR-based generation of DNA for mRNA production; (3) expression vectors for large-scale protein production; and (4) a translation reaction that does not require a membrane. The combination of these elemental processes with robotic automation resulted in high-throughput protein synthesis.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.