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Figure 9 | Microbial Cell Factories

Figure 9

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 9

Comparison of cytoplasmic and periplasmic soluble expression Genes of scFv225.28S and its mutant R4.1 were cloned in plasmid pAB1 [28]. In this plasmid, scFv genes are expressed under the control of the lac promoter with the pel B signal sequence fused at their N-terminal extremity in order to target protein to the periplasm. Soluble and insoluble extracts were prepared from strain TG1 and were analyzed by Coomassie blue staining (A) and western blot (B) as in Figure 8. The two last extracts (Cyto) are those analyzed in lanes 1 and 2 of Figure 8 (soluble cytoplasmic extracts of scFv225.28S and R4.1, cloned in pPM210 and expressed in TG1lon).

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Microbial Cell Factories

ISSN: 1475-2859

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