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Updated: Feb 12, 2026

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
Published on: January 6, 2015
SlyD-deficient Escherichia coli strains: A highway to contaminant-free protein extraction
Vladislav V Mokhonov1, Ekaterina A Vasilenko1, Ekaterina N Gorshkova1
1Center of Molecular Biology and Biomedicine, Institute of Biology and Biomedicine, Lobachevsky State University, Nizhniy Novgorod, Russia.
Abstract:
Binding of native bacterial protein SlyD to metal affinity matrices remains a major problem in affinity purification of His-tagged recombinant proteins from Escherichia coli cells. In this study, four novel E. coli strains that lack the expression of SlyD/SlyX, were engineered using λ-red mediated chromosomal deletion. The resultant mutant E. coli strains allow us to obtain SlyD-free proteins immediately after metal affinity chromatography, and eliminate additional purification processes. As a model protein, bispecific antibodies composed of anti-F4/80 VHH module and anti-TNF VHH module (MYSTI-2) were used. Using this protein we have shown that the SlyD/SlyX-deficient E. coli strains allow us to obtain a fully functional protein.
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