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Asparaginase II-GFP fusion as a tool for studying the secretion of the enzyme under nitrogen starvation
Provedor de dados: 58
Autores: Sotero-Martins,Adriana; Bon,Elba Pinto da Silva; Carvajal,Elvira.
Production of asparaginase II of Saccharomyces cerevisiae is regulated by nitrogen and can be used as a model system for studying other secreted proteins in yeast. Green fluorescent protein (GFP) from Aequorea victoria was fused to the carboxy-terminus of the enzyme by genomic integration to the locus ASP3 of S. cerevisiae. We determined asparaginase II activity, mRNA ASP3, mRNA ASP3-GFP and GFP fluorescence. Nitrogen starvation in cells carrying the chimera ASP3-GFP caused an increase in fluorescence and in the expression of ASP3. We have shown that cells producing the chimera Asp3-GFPp displayed the same response to nitrogen starvation as control cells. We demonstrated that Asp3-GFPp can be used for studying asparaginase II secretion under nitrogen...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Asparaginase II; Nitrogen regulation; Saccharomyces cerevisiae; Genomic integration; Green fluorescent protein.
Ano: 2003
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000400017