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Development of Fluorescent Biosensors for In Situ Trace-Metal Quantification and Biomonitoring

Objectives: To develop a fluorescent heavy metal biosensor that can be expressed in microalgae or immobilized on a glass surface for quantification of heavy metals in the environment.  Rationale:  There is a need for a simple, rapid, non-invasive, non-destructive and quantitative bioassay system for quantifying heavy metals in the environment.  Methodology:  A modified cyan fluorescent protein linked to a metallothionein II and a yellow fluorescent protein (Chameleon-MT) will be expressed in Chlamydomonas cells or E. coli for use as a biosensor. Upon binding Hg, Cu, Cd, Pb, Zn or Ag the Chameleon-MT will undergo fluorescence resonance energy transfer and a metal-dependent and quantifiable shift in its fluorescence emission spectrum. Differential patterns of Chameleon-MT expression will allow us to monitor heavy metals on cell surfaces as well as in the cytoplasm. In addition, we will alter the specificity and affinity of the fluorescent heavy metal biosensor. Finally, we will test hand held detection systems for field use.

Status
In progress
Type
Project
Start Date
End Date
Scope of Study
Laboratory Investigation
Scale of Phenomena
Cellular
Physical/Chemical
State Province
Ohio
Annex Numbers
Surveillance and Monitoring

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