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Molecular cloning of the imidazoleglycerolphosphate dehydratase gene of Trichoderma harzianum by genetic complementation in Saccharomyces cerevisiae using a direct expression vector

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Summary

The Trichoderma harzianum imidazoleglycerolphosphate dehydratase gene (igh) has been isolated by complementation of a Saccharomyces cerevisiae his3 mutant using a direct expression vector. This Escherichia coli-yeast shuttle vector was developed to allow efficient cloning and expression of cDNA libraries. The cDNA is 627 nucleotides long and codes for a protein of 209 amino acids with an apparent molecular mass of 22 466 daltons. The predicted protein sequence showed 63.6%, 58.7%, and 38.4% identity respectively to the corresponding enzymes from S. cerevisiae, Pichia pastoris and E. coli. Northern analysis showed that the expression of the igh gene in T. harzianum is not inhibited by external histidine and the level of igh mRNA was about threefold higher in cells starved of histidine.

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Author notes

  1. A. Herrera-Estrella

    Present address: Centro de Investigación y Estudios Avanzados, Unidad Irapuato, A.P. 629, Irapuato, Mexico

Authors and Affiliations

  1. Laboratorium voor Genetica, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000, Gent, Belgium

    G. H. Goldman, A. Herrera-Estrella, R. A. Geremia & M. Van Montagu

  2. Laboratorium voor Moleculaire Biologic, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000, Gent, Belgium

    J. Demolder, S. Dewaele & R. Contreras

Authors
  1. G. H. Goldman
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  2. J. Demolder
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  3. S. Dewaele
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  4. A. Herrera-Estrella
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  5. R. A. Geremia
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  6. M. Van Montagu
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  7. R. Contreras
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Goldman, G.H., Demolder, J., Dewaele, S. et al. Molecular cloning of the imidazoleglycerolphosphate dehydratase gene of Trichoderma harzianum by genetic complementation in Saccharomyces cerevisiae using a direct expression vector. Molec. Gen. Genet. 234, 481–488 (1992). https://doi.org/10.1007/BF00538709

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  • DOI: https://doi.org/10.1007/BF00538709

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