Asparaginase II-GFP fusion as a tool for studying the secretion of the enzyme under nitrogen starvation

Brazilian Journal of Microbiology

Adriana Sotero-martins²  Elba Pinto Da Silva Bon²  Elvira Carvajal¹ 

[1] ,Universidade Federal do Rio de JaneiroRio de Janeiro RJ ,Brasil

关键词: asparaginase II;    nitrogen regulation;    Saccharomyces cerevisiae;    genomic integration;    green fluorescent protein;    asparaginase II;    regulação por nitrogênio;    Saccharomyces cerevisiae;    integração genômica;    proteína de fluorescência verde;   

DOI  :  10.1590/S1517-83822003000400017

来源: SciELO

【 摘 要 】

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 starvation in vivo.

【 授权许可】

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