Isolation and identification of the Aspergillus nidulans gdhA gene encoding NADP-linked glutamate dehydrogenase |
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| Authors: | Sarah Jane Guff Alastair R. Hawkins Constantin Drainas James R. Kinghorn |
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| Affiliation: | (1) Department of Biochemistry and Microbiology, University of St. Andrews, Irvine Building, North Street, St. Andrews Fife, UK;(2) Department of Biochemistry, University of Leicester, Adrian Building, Leicester, UK;(3) Department of Chemistry, University of Ioannina, Ioannina, Greece;(4) Present address: Department of Genetics, University of Newcastle, Ridley Building, Newcastle, UK |
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| Abstract: | Summary The Neurospora crassa am gene was used as a heterologous probe to identify clones from two independently constructed Aspergillus nidulans gene libraries. These clones have a common HindIII 1.85 kb fragment. This A. nidulans nucleotide stretch hybridises to a N. crassa 2.7 kb BamHI fragment of wild type DNA but not to a co-migrating fragment from the DNA of the N. crassa am132 deletion mutant. One A. nidulans clone was shown to complement the N. crasse am132 deletion strain. The N. crassa transformants show low levels ( 5%) of heterologous glutamate dehydrogenase activity. The A. nidulans gdhA gene was found to locate in N. crassa at both the  homologous (i.e. am) site as well as non-nomologous sites. Partial nucleotide analysis of the fragment has revealed the 5 end of the locus and considerable homology with the N. crassa am gene. We concluded that we have cloned the A. nidulans gdhA gene.Abbreviations bp base pairs - kb 1,000 bp - GDH NADP-Iinked glutamate dehydrogenase - NADP nicotinamide adenine dinuc leotide phosphate - SDS sodium dodecyl sulfate -  phage lambda - DTT dithiothreitol - SSC 0.15 M NaCl, 0.015 Na3 citrate pH 7.0 |
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| Keywords: | Aspergillus nidulans Glutamate dehydrogenase Ammonium assimilation Fungal genes |
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