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Targeted integration into the Acremonium chrysogenum genome: disruption of the pcbC gene

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Abstract

The cephalosporin C-producing fungus Acremonium chrysogenum was transformed to hygromycin B resistance using different vector constructs. These constructs contain sequences of the pcbC gene from A. chrysogenum, encoding isopenicillin N synthetase. Detailed analysis of transformants, including pulsed-field gel electrophoresis (PFGE), suggests that integration of multiple vector copies takes place predominantly via non-homologous integration. By increasing the length of vector-DNA homologous to genomic DNA, integration occurs more frequently into chromosome VI, carrying the endogencous pcbC gene copy. In gene disruption experiments, the length of vector homology required to obtain cephalosporin C-minus transformants was investigated. Inactivation of the pcbC gene was observed only when homologous fragments of more than 3.0 kb were used on both sites of the resistance cassette. Southern analysis indicated homologous, as well as heterologous, integration of recombinant DNA. The integration of multiple vector copies leads to the appearance of truncated pcbC transcripts.

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Authors and Affiliations

  1. Lehrstuhl für Allgemeine Botanik, Ruhr-Universität Bochum, D-44780, Bochum, Germany

    Markus Walz & Ulrich Kück

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  1. Markus Walz
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  2. Ulrich Kück
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Communicated by K. Wolf

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Walz, M., Kück, U. Targeted integration into the Acremonium chrysogenum genome: disruption of the pcbC gene. Curr Genet 24, 421–427 (1993). https://doi.org/10.1007/BF00351851

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  • DOI: https://doi.org/10.1007/BF00351851

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