Methods in molecular biology · 2011 · 43 citations · 20 references
Life SciencesBiochemistryMolecular BiologyProtein EngineeringProtein TranslationCell-free Synthetic BiologyProtein Expression EngineeringProtein SynthesisCell-free Metabolic EngineeringCell-free Transcription-translation SystemsCell-free Translation Systems
We describe a system for the cell-free expression of proteins based on extracts from Escherichia coli. Two reaction configurations, batch and continuous exchange, are discussed and analytical scale as well as preparative scale setups are documented. Guidelines for the systematic development and optimization of cell-free expression protocols are given in detail. We further provide specific protocols and parameters for the cell-free production of membrane proteins. High-throughput screening applications of CF expression systems are exemplified as new tools for genomics and proteomics studies.
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Species-independent translational leaders facilitate cell-free expression
Sergey Mureev, Oleksiy Kovtun, Uyen Nguyen et al. · Nature Biotechnology · 2009 · 150 citations · Full text
Cell‐free synthesis of functional aquaporin Z in synthetic liposomes
Norman Hovijitra, Jessica J. Wuu, Boris Peaker et al. · Biotechnology and Bioengineering · 2009 · 114 citations