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Fungal Genet Biol ; 30(2): 105-13, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11017766

RESUMO

The uapC gene of Aspergillus nidulans belongs to a family of nucleobase-specific transporters conserved in prokaryotic and eucaryotic organisms. We report the use of immunological and green fluorescent protein based strategies to study protein expression and subcellular distribution of UapC. A chimeric protein containing a plant-adapted green fluorescent protein (sGFP) fused to the C-terminus of UapC was shown to be functional in vivo, as it complements a triple mutant (i.e., uapC(-) uapA(-) azgA(-)) unable to grow on uric acid as the sole nitrogen source. UapC-GFP is located in the plasma membrane and, secondarily, in internal structures observed as fluorescent dots. A strong correlation was found between cellular levels of UapC-GFP fluorescence and known patterns of uapC gene expression. This work represents the first in vivo study of protein expression and subcellular localization of a filamentous fungal nucleobase transporter.


Assuntos
Aspergillus nidulans/metabolismo , Proteínas Fúngicas , Proteínas de Membrana Transportadoras , Proteínas Recombinantes de Fusão/metabolismo , Frações Subcelulares/metabolismo , Aspergillus nidulans/genética , Aspergillus nidulans/crescimento & desenvolvimento , Membrana Celular/metabolismo , Fluorometria , Proteínas de Fluorescência Verde , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Microscopia de Fluorescência , Testes de Precipitina , Proteínas Recombinantes de Fusão/genética
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