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:
- Bioluminescent Bioreporter Integrated Circuits
- Pathogen detection using engineered bacteriophage
- Development of Noninvasive Bioluminescence Imaging for Cancer Diagnosis and Therapeutic Testing
- Detection of Estrogens by Bioluminescent Mammalian Cells
- Detection of hormonally active agents in the environment using yeast bioreporters
- Investigation of vitellogenin gene expression and development of transgenic zebrafish as bioreporters of estrogenic substances
- Codon Optimization and Expression of the
Photorhabdus luminescence luxAB genes in
Candida albicans
For more information, contact Gary Sayler or John Sanseverino.
Current Research
- Bioluminescent Bioreporter Integrated Circuits
- Pathogen detection using engineered bacteriophage
- Development of Noninvasive Bioluminescence Imaging for Cancer Diagnosis and Therapeutic Testing
- Detection of Estrogens by Bioluminescent Mammalian Cells
- Detection of hormonally active agents in the environment using yeast bioreporters
- Investigation of vitellogenin gene expression and development of transgenic zebrafish as bioreporters of estrogenic substances
- Codon Optimization and Expression of the Photorhabdus luminescence luxAB genes in Candida albicans
Figure 1. Bioluminescence emitted from individual colonies of microbial cells containing the genes for bacterial luciferase.


