<i>Escherichia coli</i> amino acid auxotrophic expression host strains for investigating protein structure–function relationships
Toshio Iwasaki, Yoshiharu Miyajima-Nakano, Risako Fukazawa, Myat T. Lin, Shinichi Matsushita, Emi Hagiuda, Alexander T. Taguchi, Sergei A. Dikanov, Yumiko Oishi, Robert B. Gennis
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.