Optimizing Membrane Protein Overexpression in the Escherichia coli strain Lemo21(DE3)

Bacteriophage membrane protein P9 as a fusion partner for the efficient expression of membrane proteins in Escherichia coli

“…Researchers have also attempted to modify host bacterial cells to optimize membrane protein production. By tuning the transcription rate of target genes and reducing the adverse effects of overexpression of membrane proteins, Schlegel et al optimized an E. coli host system for efficient membrane protein expression [15,16]. Massey-Gendel et al developed a method to select mutant strains of E. coli that improve the expression of target membrane proteins and demonstrated high levels of expression of membrane proteins from Mycobacterium tuberculosis in these mutant strains [17].…”

Engineering Escherichia coli for Functional Expression of Membrane Proteins

“…Moreover, the fluorescence reports the absolute amount of folded fusion protein, which can be observed not only in gel but also in vivo Linares et al, 2010). We and others have observed that the in vivo activity of transport proteins correlates with the corresponding in-cell and in-gel GFP fluorescence of the fusion proteins (Geertsma, Groeneveld, et al, 2008;Hibi et al, 2008;Schlegel et al, 2012). In brief, the glutamate transporter GltP from E. coli and lactose transporter LacSΔIIA from Streptococcus thermophilus have been fused to the N-terminus of GFP and expressed in E. coli.…”

The synthesis of recombinant membrane proteins in yeast for structural studies

“…However, high mRNA synthesis rates may be countered by high rates of mRNA degradation [19]. Moreover, evidence from prokaryotic expression systems suggests that acquired mutations that lower promoter efficiency lead to improved functional yields of membrane proteins for some, but not all, targets [20]. In a separate study, a series of E. coli strains that had been evolved to improve their yield characteristics were found to have a mutation in the hns gene, which has a role in transcriptional silencing [21].…”