Physiological Testing of Host Pathogens in an
E. coli Specific Bioreporter System
Bioreporter Mechanism
In the closed environment of the space shuttle, there is great potential for pathogenic activity to occur. With this bioreporter system, we will be able to monitor pathogenic activity by creating a photoreceptor able to measure bioluminescence produced due to interactions between several genes.
By conducting physiological tests on the host cells we are able to figure out whether the bioreporter system can function when the cells are stressed.
Genes and their Functions
• Lambda phage — E. coli specific
Contains luxI gene
• Phage infects host pathogen, integrating luxI gene
luxI produces acetyl homoserine lactone (AHL) molecules
• Bioreporter contains luxR gene
luxR recognizes AHL molecules and initiates lux cassette to produce bioluminescence
Physiological Testing and Outcomes
• Starvation of host pathogens
• 1 hour and 1 day
• Count numbers of cells that are living and dead by staining
• Measure amount of light produced by stressed cells using a 96 well plate and plate reader
• Based on the data collected, stress on the host cells does not decrease the amount of light produced within the bioreporter system.
• Time for peak light produced increases, while intensity of light remains the same
Author: Carmen Pearson, North Carolina State University
Principal Investigators: Kathleen Daumer, Dynamac Corporation
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