1. Cloning of the regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans
- Author
-
Brownlee Ag, Johnson Ri, Taylor Lh, Mark X. Caddick, and Herbert N. Arst
- Subjects
Genetics ,General Immunology and Microbiology ,biology ,Nitrogen ,General Neuroscience ,Genes, Fungal ,Mutant ,Nucleic Acid Hybridization ,DNA Restriction Enzymes ,Molecular cloning ,biology.organism_classification ,Molecular biology ,Aspergillus nidulans ,General Biochemistry, Genetics and Molecular Biology ,Genes, Regulator ,Genomic library ,Cloning, Molecular ,Molecular Biology ,Gene ,Research Article ,Southern blot ,Regulator gene ,Chromosomal inversion - Abstract
The areA gene, which mediates nitrogen metabolite repression in the fungus Aspergillus nidulans, lies sufficiently close to a telomere that no indispensable gene can be distal to it. We were able therefore to exploit the existence of a near terminal pericentric inversion to devise a method for cloning areA plus the region beyond it towards the telomere. In crosses heterozygous for this inversion a class of duplication-deficient progeny lacking areA and the region centromere-distal to it is obtained. We, therefore, sought clones from an A. nidulans gene library in lambda Charon 4 able to hybridize to total genomic DNA from a wild-type strain but not to that from a duplication-deficiency strain. A clone, containing an 11.6-kb insert, which hybridised weakly to duplication-deficiency DNA, overlapped chromosome breakpoints of three different aberration-associated areA alleles and was able to transform an areA mutant to areA+. Southern blotting and genetic analysis established that the transforming sequence had integrated in the region centromere distal to areA. The cloning method yielded other clones from the region centromere-distal to areA which were used to show that the translocation associated with a mutant areA allele is reciprocal rather than non-reciprocal, a fact which could not be established by classical genetics. Finally, analysis of the cloned portion of the dispensable region centromere-distal to areA indicates that this region contains at least 0.5% of the A. nidulans genome.
- Published
- 1986
- Full Text
- View/download PDF