Engineering in Life Sciences · 2012 · 72 citations · 38 references
Life SciencesBiochemistryMolecular BiologyPost-translational ModificationProtein EngineeringProtein TranslationCell-free Synthetic BiologyMembrane Protein EngineeringMembrane ProteinsProtein Expression EngineeringProtein SynthesisTranslation In EukaryotesCell-free Translation SystemsProtein Overexpression
Cell‐free protein synthesis (CFPS) is a valuable method for the fast expression of difficult‐to‐express proteins as well as posttranslationally modified proteins. Since cell‐free systems circumvent possible cytotoxic effects caused by protein overexpression in living cells, they significantly enlarge the scale and variety of proteins that can be characterized. We demonstrate the high potential of eukaryotic CFPS to express various types of membrane proteins covering a broad range of structurally and functionally diverse proteins. Our eukaryotic cell‐free translation systems are capable to provide high molecular weight membrane proteins, fluorescent‐labeled membrane proteins, as well as posttranslationally modified proteins for further downstream analysis.
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Phosphorylation of initiation factor eIF-2 and the control of reticulocyte protein synthesis
Paul J. Farrell, Ken Balkow, Tim Hunt et al. · Cell · 1977 · 752 citations