Fluorescence-based sensing system for copper using genetically engineered living yeast cells

Ranjit S. Shetty, Sapna K. Deo, Yue Liu, Sylvia Daunert

Research output: Contribution to journalArticle

40 Scopus citations

Abstract

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 Acel present in S. cerevisiae to control the expression of the reporter protein, GFPuv. When copper ions are present in the sample, the Acel protein activates the cupl promoter located upstream from the gfpuv 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 x 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.

Original languageEnglish (US)
Pages (from-to)664-670
Number of pages7
JournalBiotechnology and Bioengineering
Volume88
Issue number5
DOIs
StatePublished - Dec 5 2004
Externally publishedYes

Keywords

  • Copper detection
  • GFPuv
  • Reporter gene assay
  • Whole cell sensing

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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