Recombinant protein expression in Escherichia coli

Current Opinion in Biotechnology

Volumn 10, Issue 5, 1999, Pages 411-421

  Baneyx, François ^a  

^a University of Washington   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

RECOMBINANT PROTEIN;

BACTERIAL GENETICS; ESCHERICHIA COLI; EUKARYOTE; EXPRESSION VECTOR; GENETIC TRANSCRIPTION; NONHUMAN; PRIORITY JOURNAL; PROTEIN EXPRESSION; PROTEIN FOLDING; REVIEW; RNA TRANSLATION;

ESCHERICHIA COLI;

EID: 0032884573     PISSN: 09581669     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0958-1669(99)00003-8     Document Type: Review

References (69)

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66. Hayhurst A, Harris WJ Escherichia coli Skp chaperone coexpression improves solubility and phage display of single chain antibody fragments. Protein Expr Purif. 15:1999;336-343. This study shows that Skp coexpression can greatly improve the solubility of a highly toxic ScFv secreted in the periplasm and allow its phage display. Skp accumulation is higher when the protein is synthesized from a dicistronic operon than when provided on compatible plasmids under ara transcriptional control. High levels of Skp correlate with increase ScFv solubility.
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69. Wan EW, Baneyx F TolAIII co-overexpression facilitates the recovery of periplasmic recombinant proteins into the growth medium of Escherichia coli. Protein Expr Purif. 14:1998;13-22. E. coli strains bearing mutations in the tolQRAB gene cluster or certain other cell envelope proteins spontaneously release periplasmic proteins in the extracellular medium. Unfortunately, these bacteria are fragile and not amenable to high density fermentation. In this study, a ColE1-compatible plasmid encoding the third topological domain of TolA (TolAIII) is used to mimic the 'leaky' phenotype of tolA mutants. The usefulness of TolAIII coexpression in promoting the efficient release of resident and recombinant periplasmic proteins is demonstrated. It is noted that although this strategy reduces the total levels of recombinant β-lactamase secreted from the ompA leader, formation of periplasmic inclusion bodies is completely abolished.