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Yeast exo‐β‐glucanases can be used as efficient and readily detectable reporter genes in Saccharomyces cerevisiae
Author(s) -
Cid V. J.,
Alvarez A. M.,
Santos A. I.,
Nombela C.,
Sanchez M.
Publication year - 1994
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320100606
Subject(s) - saccharomyces cerevisiae , biology , reporter gene , yeast , plasmid , beta galactosidase , candida albicans , flow cytometry , gene , cell sorting , microbiology and biotechnology , genetics , gene expression
Yeast exo‐1,3‐β‐glucanases are secretable proteins whose function is basically trophic and may also be involved in cell wall glucan hydrolytic processes. Since fluorescein di(β‐ D ‐glucopyranoside) is a fluorogenic substrate detectable and quantifiable by flow cytometry, it was used for testing the ability of the EXG1 gene product of Saccharomyces cerevisiae and its homologous gene in Candida albicans to function as reporter genes. These open reading frames were coupled to different promoters in multicopy plasmids, and exoglucanase activity quantified at flow cytometry. Exoglucanases were found to be useful tools for the study of promoter regions in S. cerevisiae . This technique has the advantage over other reporter gene systems—such as β‐galactosidase fusions—that it does not require permeabilization of yeast cells and therefore it allows the recovery of viable cells—by sorting—after flow cytometry analysis.