Cloning of the Aspergillus oryzae 5-aminolevulinate synthase gene and its use as a selectable marker |
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Authors: | Susan L Elrod Aubrey Jones Randy M Berka Joel R Cherry |
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Institution: | (1) Novo Nordisk Biotech Inc., 1445 Drew Avenue, Davis, CA 95616, USA e-mail: selrod@calpoly.edu, US |
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Abstract: | The hemA gene encoding 5-aminolevulinate synthase, the first enzyme in heme biosynthesis, was cloned from Aspergillus oryzae and evaluated as a selectable marker for the transformation of filamentous fungi. Deletion of the hemA gene resulted in a lethal phenotype that could be rescued either by the supplementation of culture media with 5-aminolevulinic
acid (ALA) or by transformation with the wild-type hemA gene, but not by growth on rich media, nor by the addition of exogenous heme. Transformation of a hemA deletion strain with the hemA gene linked to a lipase expression cassette yielded ALA prototrophs expressing lipase. The hemA gene can therefore be used as a selectable marker for the transformation of A. oryzae.
Received: 16 March 2000 / Accepted: 18 July 2000 |
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Keywords: | hemA Heterologous gene expression Heme biosynthesis Recombinant enzyme |
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