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Appl Environ Microbiol. 1992 December; 58(12): 4038-4041

High-level expression of the Streptomyces clavuligerus isopenicillin N synthase gene in Escherichia coli.

Department 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.

This article has been cited by other articles:

• Alexander, D. C., Jensen, S. E. (1998). Investigation of the Streptomyces clavuligerus Cephamycin C Gene Cluster and Its Regulation by the CcaR Protein. J. Bacteriol. 180: 4068-4079 [Abstract] [Full Text]