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Figure 3 | Microbial Cell Factories

Figure 3

From: Directed evolution of single-chain Fv for cytoplasmic expression using the β-galactosidase complementation assay results in proteins highly susceptible to protease degradation and aggregation

Figure 3

Effect of the 30 aminoacid long C-terminal peptide on scFv expression level scFv13 and scFv13R4 were cloned either in pPM170 (173 and 173R4) or in pPM173S1 (173S1 and 173S1-R4), and expressed in strain TG1. A) Phenotype of the strains on MacConkey lactose plates after 24 h at 37°C. B) Soluble and insoluble extracts of the four strains were analyzed by Coomassie blue staining (top) or by western blot (bottom) using the 9E10 anti-c-myc monoclonal antibody followed by an anti-mouse HRP-conjugated antibody and detected with a commercial kit (Pierce Supersignal West Pico kit #14079). The arrows show the position of the fusions (38 kDa for 173 and 173R4; 34 kDa for 173S1 and 173S1-R4) and the unfused scFv (28 kDa).

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Microbial Cell Factories

ISSN: 1475-2859

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