Báo cáo khoa học: Optimization of an Escherichia coli system for cell-free synthesis of selectively 15N-labelled proteins for rapid analysis by NMR spectroscopy
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Cell-free protein synthesis offers rapidaccess toproteins that are selectively labelledwith [ 15 N]aminoacids andsuitable for analysis by NMR spectroscopy without chromatographic purification. A system based on anEscherichia colicell ex-tractwas optimizedwith regard toproteinyieldandminimal usage of 15 N-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 reactionmedium were obtained by expression of a synthetic gene. ...
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