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PMID:1743513
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| Citation |
Gianì, S, Manoni, M and Breviario, D (1991) Cloning and transcriptional analysis of the ADE6 gene of Saccharomyces cerevisiae. Gene 107:149-54 |
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| Abstract |
The Saccharomyces cerevisiae gene, ADE6, encoding 5'-phosphoribosylformyl glycinamidine synthetase (EC 6.3.5.3) has been cloned by complementation of an ade6 auxotroph. Transformation of ade6 mutants with ADE6-carrying centromeric plasmids restored normal, adenine-independent growth behavior in the recipients. Strains containing a disrupted ade6 allele were constructed and behaved as stable adenine auxotrophs. Southern transfer and genetic analyses of strains carrying a disrupted ade6 allele demonstrated that the cloned gene was ADE6 and not a suppressor. The cloned ADE6 DNA was mapped on the RAD2-proximal fragment of chromosome VII by hybridization on yeast chromosomes separated by pulsed-field gel electrophoresis. Northern-blot hybridization experiments show that the ADE6 region produces two different mRNA species of approx. 5 and 2 kb. Disappearance of the larger, but not the smaller, transcript is associated with ade6 mutations. A threefold repression in the amount of the 5-kb ADE6 mRNA is observed when growth medium is supplemented with exogenous adenine. |
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| Keywords |
Adenine/pharmacology; Blotting, Northern; Blotting, Southern; Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; Cloning, Molecular; Electrophoresis, Gel, Pulsed-Field; Gene Expression Regulation, Fungal/genetics; Genes, Fungal/genetics; Ligases/genetics; Plasmids/genetics; Restriction Mapping; Saccharomyces cerevisiae/drug effects; Saccharomyces cerevisiae/enzymology; Saccharomyces cerevisiae/genetics; Transcription, Genetic/genetics; Transformation, Genetic/genetics |
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