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Ohio Sea Grant College Program
and Stone Laboratory

Ohio Sea Grant and Stone Laboratory

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

Project Number: R/BT-008, Progress Report

Start Date: 6/1/2003

Completion Date: 7/31/2007

Revision Date: 8/24/2006

Principal Investigator(s)1.Richard T. Sayre, Biochemistry and Plant Biology The Ohio State University*
This shows the current affiliation and may not match affiliation at time of participation. *

Funding Record

Source: Ohio Sea Grant College Program
Source FundState MatchPass Through
Total$ 255,882.00$ 148,489.00$ 0.00

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.

Publications & Media

Presentations
Presentations• Rajamani S, Ewalt J, Torres M, Falcao V, Colepicolo P and Sayre RT 2006, Transgenic Microalgae as Heavy Metal Biosensors.
ASLO/AGU Ocean Sciences Meeting, Honolulu, HI.
Presentations• Sayre RT, Rajamani S, Falcao V, Torres M, Ewalt J and Colepicolo P 2005, Microalgae as Heavy Metal Biosensors.
International Marine Biotechnology Conference. St Johns, Newfoundland, Canada. Third Place Poster Award

Supported Students

StudentSathish Rajamani (Graduate, Ph. D.)
Ohio State University
StudentZoee Gokhale (Graduate, Ph. D.)
Ohio State University