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Improved yeast-screening assay for phytoestrogens

Biosci Biotechnol Biochem. 2021-03; 
Natthida Sophon, Choladda Phiwpesh, Sieng Darith, Wichai Cherdshewasart, Chuenchit Boonchird
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Plasmid DNA Preparation … The plasmid pUC57 harboring synthetic yEGFP (Cormack et al. 1997) was purchased from GenScript™ (USA). The YEp type plasmid pYES2 with URA3 selectable markers was obtained from Invitrogen™ (USA) and applied for construction of yEGFP reporter plasmid … Get A Quote

摘要

A yeast estrogen screening (YES) assay was improved to increase sensitivity for detection of phytoestrogens. New yeast strains minus one or the other of transporters Pdr5 or Snq2 and harboring yEGFP as a reporter gene were developed. The new strains showed 2-100 fold improvement in sensitivity for detection of standard estrogens and anti-estrogens. In addition, the assay time (1h) using the newly developed strains was shorter than that (4h) previously reported. Furthermore, the snq2-minus strains were most effective for detection of estrogenic activity while the pdr5-minus strains were most effective for detection of anti-estrogenic activity. The efficacy of the new methods was evaluated and confirmed by testin... More

关键词

PDR5, SNQ2, YES assay, phytoestrogens, yeast enhanced green fluorescent protein