A chimeric light-regulated amino acid transport system allows the isolation of blue light regulator (blr) mutants of Neurospora crassa
| Autor: | M Rodriguez-Franco, Alessandra Carattoli, Giuseppe Macino, E Kato, W D Stuart |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
southern blotting
Amino Acid Transport Systems Light White Collar-1 Messenger Mutant 4 fluorophenylalanine carotenoid messenger rna n methyltryptophan amino acid transport article gene control gene fusion gene mutation genetic transformation neurospora crassa nonhuman northern blotting phenotype photosensitivity priority journal promoter region regulator gene southern blotting Amino Acid Transport Systems Amino Acids Base Sequence Blotting Northern Blotting Southern Carrier Proteins Chimera DNA Primers DNA Fungal Gene Expression Regulation Fungal Genes Fungal Molecular Sequence Data Mutagenesis Neurospora crassa Polymerase Chain Reaction Promoter Regions (Genetics) RNA Messenger Support Non-U.S. Gov't Ultraviolet Rays Neurospora crassa chemistry.chemical_compound Aromatic amino acids Northern Promoter Regions Genetic Non-U.S. Gov't Southern Regulator gene chemistry.chemical_classification Multidisciplinary amino acid transport biology Blotting n methyltryptophan Amino acid Amino acid permease Fungal Biochemistry Support Research Article Ultraviolet Rays Tryptophan DNA biology.organism_classification Molecular biology Gene Expression Regulation Genes chemistry RNA |
| Zdroj: | Proceedings of the National Academy of Sciences. 92:6612-6616 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.92.14.6612 |
| Popis: | We have developed a system for the isolation of Neurospora crassa mutants that shows altered responses to blue light. To this end we have used the light-regulated promoter of the albino-3 gene fused to the neutral amino acid permease gene mtr. The product of the mtr gene is required for the uptake of neutral aliphatic and aromatic amino acids, as well as toxic analogs such as p-flurophenylalanine or 4-methyltryptophan. mtr trp-2-carrying cells were transformed with the al-3 promoter-mtr wild-type gene (al-3p-mtr+) to obtain a strain with a light-regulated tryptophan uptake. This strain is sensitive to p-fluorophenylalanine when grown under illumination and resistant when grown in the dark. UV mutagenesis of the al-3p-mtr(+)-carrying strain allowed us to isolate two mutant strains, BLR-1 and BLR-2 (blue light regulator), that are light-resistant to p-fluorophenylalanine and have lost the ability to grow on tryptophan. These two strains have a pale-orange phenotype and show down-regulation of all the photoregulated genes tested (al-3, al-1, con-8, and con-10). Mutations in the BLR strains are not allelic with white collar 1 or white collar 2, regulatory genes that are also involved in the response to blue light. |
| Databáze: | OpenAIRE |
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