Cloning of the regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans
| Autor: | Brownlee Ag, Johnson Ri, Taylor Lh, Mark X. Caddick, Herbert N. Arst |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
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 |
| Zdroj: | The EMBO Journal. 5:1087-1090 |
| ISSN: | 0261-4189 |
| DOI: | 10.1002/j.1460-2075.1986.tb04326.x |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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