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Optimization of an Escherichia coli system for cell-free synthesis of selectively 15 N-labelled proteins for rapid analysis by NMR spectroscopy

Ozawa, Kiyoshi; Headlam, Madeleine; Schaeffer, Patrick; Henderson, Blair; Dixon, Nicholas; Otting, Gottfried

Description

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]

CollectionsANU Research Publications
Date published: 2004
Type: Journal article
URI: http://hdl.handle.net/1885/87258
Source: European Journal of Biochemistry
DOI: 10.1111/j.1432-1033.2004.04346.x

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