Streamlined β-Galactosidase Assay for Analysis of Recombinant Yeast Response to Estrogens | Zendy

Ronan Le Guével | Zendy; Farzad Pakdel | Zendy

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Streamlined β-Galactosidase Assay for Analysis of Recombinant Yeast Response to Estrogens

Author(s) -

Ronan Le Guével,

Farzad Pakdel

Publication year - 2001

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/01305st05

Subject(s) - yeast , recombinant dna , lysis , arthrobacter , biology , bioassay , saccharomyces cerevisiae , biochemistry , microbiology and biotechnology , enzyme , gene , genetics

Here, we describe a rapid, convenient, and quantitative beta-galactosidase assay in liquid culture of recombinant yeast that expresses the estrogen receptor. This assay allows large-scale screening of chemicals (more than 600 samples/day) for the evaluation of their direct estrogenic potency and accurate determination of their EC50 with minimal manipulations. The assay, which is based on digestion of the yeast cell wall by lyticase (zymolase), a beta-glucanase isolated from Arthrobacter luteus, followed by a hypoosmotic shock lysis, is performed completely in 96-well plates. This protocol for using recombinant yeast with the two-hybrid technology significantly advances recombinant yeast manipulation.

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