Optimization of an Escherichia coli system for cell-free synthesis of selectively 15 N-labelled proteins for rapid analysis by NMR spectroscopy
Cell-free protein synthesis offers rapid access to proteins that are selectively labelled with [15N]amino acids and suitable for analysis by NMR spectroscopy without chromatographic purification. A system based on an Escherichia coli cell extract was optimized with regard to protein yield and minimal usage of 15N-labelled amino acid, and examined for the presence of metabolic by-products which could interfere with the NMR analysis. Yields of up to 1.8 mg of human cyclophilin A per mL of...[Show more]
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|Source:||European Journal of Biochemistry|
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