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The University of Tennessee

Center for Environmental Biotechnology

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Research » Bioreporter Development and Bioimaging


Bioreporter Development and Bioimaging

Bioluminescent bioreporters are intact, living microorganisms genetically designed to respond to specific chemical or physical agents in their environment via the production of visible light. These bioreporters use the lux gene cassette derived from naturally bioluminescent bacteria such as Vibrio fischeri or Photorhabdus luminescens. The lux cassette consists of five genes, luxA, B, C, D, and E. The luciferase genes (luxAB) encode for the proteins responsible for generating bioluminescence while the reductase (luxC), transferase (luxD), and synthetase (luxE) genes encode for proteins involved in producing an aldehyde substrate required in the bioluminescent reaction. With all five of these genes operating in concert, the bioreporter cell is able to generate a fully self-directed bioluminescent signal in response to specific chemical or physical agent exposures, thus, no user interaction is required once the assay has been initiated. The intensity of the bioluminescent response then correlates to the concentration of the specific exposure agent, thereby reporting on the target’s presence as well as its bioavailability and overall effect on a living system. Due to their small size, bioreporters can also efficiently integrate into existing lab-on-a-chip and microfluidic technology interfaces to yield continuous, on-line, and near real-time detection and monitoring devices.

The Center for Environmental Biotechnology has been a leader in utilizing bacterial bioluminescence for environmental sensing from development of one of the first bioreporters for monitoring naphthalene bioavailability (King et al., 1990) to the first field release of a genetically modified microorganism for bioremediation applications (Ripp et al., 2000). Recent research has focused on developing bioluminescent bioreporter integrated circuits (BBIC); devices that fuse bacterial bioreporters with photodetectors on integrated circuits (Bolton et al., 2002).

CEB researchers were the first to insert the Photorhabdus luminescens lux operon into the yeast Saccharomyces cerevisiae for fully autonomous bioluminescence (Gupta et al., 2003). Bioluminescent yeast bioreporters opens a new suite of targets for environmental sensing including environmental estrogens (Sanseverino et al., 2005) and androgens (Eldridge et al., 2007). Building on the success of bacterial lux expression in lower eukaryotes, the P. luminescens lux genes have been codon optimized for expression in mammalian tissue cultures (Patterson et al., 2005). Autonomous bioluminescence expression opens a new avenue for tissue-based and whole animal bioimaging for the detection of, for example, cancer, blood glucose levels, and hormones.

The current research portfolio of CEB builds on this previous success. Current projects include:

For more information, contact Gary Sayler or John Sanseverino.