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Fluorescence‐based sensing system for copper using genetically engineered living yeast cells
Author(s) -
Shetty Ranjit S.,
Deo Sapna K.,
Liu Yue,
Daunert Sylvia
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20331
Subject(s) - green fluorescent protein , copper , saccharomyces cerevisiae , plasmid , yeast , fluorescence , activator (genetics) , metal ions in aqueous solution , biology , chemistry , reporter gene , biophysics , microbiology and biotechnology , biochemistry , gene , ion , gene expression , quantum mechanics , physics , organic chemistry
A whole cell‐based optical sensing system for copper was developed based on Saccharomyces cerevisiae cells harboring plasmid pYEX‐GFPuv. The basis of this system was the ability of the transcriptional activator protein Ace1 present in S. cerevisiae to control the expression of the reporter protein, GFPuv. When copper ions are present in the sample, the Ace1 protein activates the cup1 promoter located upstream from the gfp uv gene in plasmid pYEX‐GFPuv, thus inducing the production of GFPuv. The concentration of copper ions in the sample can then be related to the GFPuv expressed in the yeast. The amount of GFPuv produced in the system was determined by monitoring the fluorescence emitted at 507 nm after excitation at 397 nm. This system can detect copper at concentrations as low as 5 × 10 −7 M, and is selective for copper over a variety of metal ions, with the exception of silver. The applicability of this sensing system to different analytical platforms and in real samples is demonstrated. © 2004 Wiley Periodicals, Inc.