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Updated: May 6, 2026

Escherichia coli-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology
Published on: February 25, 2019
Engine out of the chassis: cell-free protein synthesis and its uses
Gabriel Rosenblum1, Barry S Cooperman1
1Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104-6323, United States.
Abstract:
The translation machinery is the engine of life. Extracting the cytoplasmic milieu from a cell affords a lysate capable of producing proteins in concentrations reaching to tens of micromolar. Such lysates, derivable from a variety of cells, allow the facile addition and subtraction of components that are directly or indirectly related to the translation machinery and/or the over-expressed protein. The flexible nature of such cell-free expression systems, when coupled with high throughput monitoring, can be especially suitable for protein engineering studies, allowing one to bypass multiple steps typically required using conventional in vivo protein expression.
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