Affiliation:
1. Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan
2. Department of Biochemistry
3. Department of Veterinary Clinical Medicine, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
Abstract
Abstract
A set of C43(DE3) and BL21(DE3) Escherichia coli host strains that are auxotrophic for various amino acids is briefly reviewed. These strains require the addition of a defined set of one or more amino acids in the growth medium, and have been specifically designed for overproduction of membrane or water-soluble proteins selectively labelled with stable isotopes, such as 2H, 13C and 15N. The strains described here are available for use and have been deposited into public strain banks. Although they cannot fully eliminate the possibility of isotope dilution and mixing, metabolic scrambling of the different amino acid types can be minimized through a careful consideration of the bacterial metabolic pathways. The use of a suitable auxotrophic expression host strain with an appropriately isotopically labelled growth medium ensures high levels of isotope labelling efficiency as well as selectivity for providing deeper insight into protein structure–function relationships.
Funder
International Collaborations in Chemistry Grant from the Japan Society for the Promotion of Science
National Science Foundation
Japan Society for the Promotion of Science
Nagase Science and Technology Foundation Research Grant
Japan Agency for Medical Research and Development Grant
Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences
United States Department of Energy, and National Institute of General Medical Sciences Roadmap Initiative
National Institutes of Health Grant
Japan Society for the Promotion of Science Postdoctoral Fellow
Publisher
Oxford University Press (OUP)
Subject
Molecular Biology,Biochemistry,General Medicine
Cited by
5 articles.
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