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Preparation of multiple site-specific mutant proteins for NMR studies by PCR-directed cell-free protein synthesis

Ozawa, Kiyoshi; Qi, Ruhu


Cell-free protein synthesis (CFPS) offers a fast and inexpensive approach to selectively label proteins with isotopes that can then be detected by nuclear magnetic resonance (NMR) spectroscopy directly in the translation mixture. We describe a PCR-based approach for production of protein-coding circularized DNA templates that can be expressed in Escherichia coli extract in CFPS dialysis system. This approach typically yields target protein concentrations close to 1 mg/mL, which is sufficient...[Show more]

CollectionsANU Research Publications
Date published: 2014
Type: Journal article
Source: Methods in Molecular Biology
DOI: 10.1007/978-1-62703-782-2_11


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