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Figure 1 | Microbial Cell Factories

Figure 1

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 1

Schematic view of plasmid pPM170 Plasmid pPM170 allows the cloning of scFv genes between the Nco I and Not I sites under the control of the lactose promoter. The translated protein is fused at its C-terminal extremity to the α-fragment of βgal. A) Plasmid map with the main restriction sites; B) DNA and aminoacid sequences fused to the 3' end and the C-terminus of the cloned scFv. The scFv sequence is followed by two tags (in blue), the α-fragment of βgal (in green, aminoacids 6 to 59 of βgal) and a 30 aminoacid long peptide (in orange) originating from pBR322 plasmid and M13 gene IV sequence (see text). The sequences of the two sequenced spontaneous lac+ mutants, 173S1 and 173S2 (see text), are also shown.

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Microbial Cell Factories

ISSN: 1475-2859

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