PMID:21964791
| Citation |
Kai, L, Roos, C, Haberstock, S, Proverbio, D, Ma, Y, Junge, F, Karbyshev, M, Dötsch, V and Bernhard, F (2012) Systems for the cell-free synthesis of proteins. Methods Mol. Biol. 800:201-25 |
|---|---|
| Abstract |
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. |
| Links |
PubMed Online version:10.1007/978-1-61779-349-3_14 |
| Keywords |
Bacteriophage T7/enzymology; Cell Fractionation; DNA/genetics; DNA/metabolism; DNA-Directed RNA Polymerases/metabolism; Detergents/chemistry; Escherichia coli/cytology; Escherichia coli/genetics; Magnesium/chemistry; Membrane Proteins/biosynthesis; Membrane Proteins/genetics; Membrane Proteins/isolation & purification; Protein Engineering/methods; Recombinant Proteins/biosynthesis; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification |
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