Abstract
The commonly used drug diclofenac is an important environmental anthropogenic pollutant. Currently, detection of diclofenac is mainly based on chemical and physical methods. Here we describe a yeast biosensor that drives the diclofenac-dependent expression of a recombinant fluorescent protein from the authentic promoter of the PDR5 gene. This key component of the pleiotropic drug response encodes a multidrug transporter that is involved in cellular detoxification. We analyse the effects on diclofenac sensitivity of artificial PDR5 promoter derivatives in wild-type and various yeast mutant strains. This approach enabled us to generate sensor strains with elevated drug sensitivity.
Highlights
Diclofenac monosodium (CAS 15307-79-6) is a non-steroidal anti-inflammatory drug (NSAID)that is prescribed against pain and rheumatic inflammations
Diclofenac-Mediated Induction of PDR5 Depends on the Transcription Factors Pdr1p and Pdr3p, the Master Regulators of the Pleiotropic Drug Response
In order to reveal the cis-acting elements and trans-acting factors that mediate this response, a reporter construct consisting of 1000 bp of the native 50 URS of PDR5 fused to TGFP was generated
Summary
Diclofenac monosodium (CAS 15307-79-6) is a non-steroidal anti-inflammatory drug (NSAID)that is prescribed against pain and rheumatic inflammations. In a long-term study of the effluents of three wastewater treatment plants (WWTP) over 10 months, diclofenac was detected with mean concentrations of 1–10 μg/L [2], whereas in a EU-wide monitoring survey of WWTP effluents a maximal concentration of 174 ng/L was detected with average values of 50 ng/L [3]. Contradictory results have been reported for the removal of diclofenac in WWTPs [4]: while in one WWTP removal rates up to 70% were found, removal was neither observed in other investigated conventional WWTPs [5] nor in a pilot sewage plant and biofilm reactors [6]. In the EU-wide monitoring survey cited above [3], analytics mainly encompassed chemical and physical methods as well as bioassays. The yeast Saccharomyces (S.) cerevisiae (strain W303.1a) was used for cytotoxicity tests based on its sensitivity to oxidative stress
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