High-level expression of the Streptomyces clavuligerus isopenicillin N synthase gene in Escherichia coli
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Updated: Oct 20, 2026

The Multifaceted Benefits of Protein Co-expression in Escherichia coli
Published on: February 5, 2015
M Durairaj1, J L Doran, S E Jensen
1Department of Microbiology, University of Alberta, Edmonton, Canada.
Abstract:
The pcbC gene, which encodes isopenicillin N synthase (IPNS), was subcloned from Streptomyces clavuligerus into Escherichia coli by using the pT7 series of plasmid vectors. The polymerase chain reaction was used to introduce an NdeI site at the translation initiation codon of pcbC, allowing the gene to be inserted behind an E. coli type of ribosome binding site. This construction directed high-level expression of IPNS, but the IPNS was in an inactive form in inclusion bodies. Active IPNS was recovered by solubilizing and renaturing the protein.
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