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Escherichia coli amino acid auxotrophic expression host strains for investigating protein structure-function relationships.
Iwasaki, Toshio; Miyajima-Nakano, Yoshiharu; Fukazawa, Risako; Lin, Myat T; Matsushita, Shin-Ichi; Hagiuda, Emi; Taguchi, Alexander T; Dikanov, Sergei A; Oishi, Yumiko; Gennis, Robert B.
Afiliación
  • Iwasaki T; Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan.
  • Miyajima-Nakano Y; Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan.
  • Fukazawa R; Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan.
  • Lin MT; Department of Biochemistry.
  • Matsushita SI; Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan.
  • Hagiuda E; Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan.
  • Taguchi AT; Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan.
  • Dikanov SA; Department of Veterinary Clinical Medicine, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
  • Oishi Y; Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Tokyo 113-8602, Japan.
  • Gennis RB; Department of Biochemistry.
J Biochem ; 169(4): 387-394, 2021 Apr 29.
Article en En | MEDLINE | ID: mdl-33289521
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Regulación Bacteriana de la Expresión Génica / Escherichia coli Idioma: En Revista: J Biochem Año: 2021 Tipo del documento: Article País de afiliación: Japón

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Regulación Bacteriana de la Expresión Génica / Escherichia coli Idioma: En Revista: J Biochem Año: 2021 Tipo del documento: Article País de afiliación: Japón