PMID: 9631547Jun 19, 1998Paper

Coupling site-directed mutagenesis with high-level expression: large scale production of mutant porins from E. coli

FEMS Microbiology Letters
A PrilipovR Koebnik

Abstract

Combination of an origin repair mutagenesis system with a new mutS host strain increased the efficiency of mutagenesis from 46% to 75% mutant clones. Overexpression with the T7 expression system afforded large quantities of proteins from mutant strains. A series of E. coli BE host strains devoid of major outer membrane proteins was constructed, facilitating the purification of mutant porins to homogeneity. This allowed preparation of 149 porin mutants in E. coli used in detailed explorations of the structure and function of this membrane protein to high resolution.

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Citations

Jun 24, 2003·Journal of Molecular Biology·Pavel JanscakThomas A Bickle
Mar 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·S R SauterV Braun
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