Isolation, characterization and disruption of the areA nitrogen regulatory gene of Gibberella fujikuroi
| Autor: | George A. Marzluf, B. Feng, Bettina Tudzynski, Veronika Homann |
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| Rok vydání: | 1999 |
| Předmět: |
Gibberella
Nitrogen Genes Fungal Molecular Sequence Data Mutant Nitrate Reductase Neurospora crassa Fungal Proteins Nitrate Reductases Gene Expression Regulation Fungal Genetics Amino Acid Sequence Cloning Molecular DNA Fungal Molecular Biology Regulator gene Zinc finger Fungal protein Sequence Homology Amino Acid biology Genetic Complementation Test food and beverages Sequence Analysis DNA biology.organism_classification Gibberellins DNA-Binding Proteins Complementation Biochemistry Gene Targeting Mutation Gibberella fujikuroi Sequence Alignment Transcription Factors |
| Zdroj: | Molecular and General Genetics MGG. 261:106-114 |
| ISSN: | 1432-1874 0026-8925 |
| DOI: | 10.1007/s004380050947 |
| Popis: | The gene areA-GF, a homologue of the major nitrogen regulatory genes nit-2, areA, nre and NUT1 of Neurospora crassa, Aspergillus nidulans, Penicillium chrysogenum and Magnaporthe grisea, respectively, was cloned from the gibberellin (GA)-producing rice pathogen Gibberella fujikuroi. areA-GF encodes a protein of 972 amino acid residues which contains a single putative zinc finger DNA-binding domain that is at least 98% identical to the zinc finger domains of the homologous fungal proteins. The areA-GF gene has been shown to be functional in N. crassa by heterologous complementation of a RIP induced nit-2 mutant. The transformation rate was nearly as high as in a homologous complementation control. Transformants were able to utilize nitrate and expressed a normally regulated nitrate reductase activity. To generate areA-GF- mutants, gene replacement experiments were performed using a linearized replacement vector carrying the hygromycin B phosphotransferase (hph) gene. The replacement of the zinc finger by the hygromycin cassette resulted in transformants which were unable to utilize nitrogen sources other than ammonium and glutamine, and gave significantly reduced gibberellin production yields. Complementation of such a mutant with the wild-type gene led to the full recovery of gibberellin production. |
| Databáze: | OpenAIRE |
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