MP1 New Approaches to Determining Soil and Sediment Exposures
256 Portland Ballroom
1:20 PM - 4:40 PM, Monday
() Direct assessment of mutagenicity using GFP transgenic C. elegans.
Glenn, T1, Graves, A2, Humphries, S3, Krizek, B3, Jagoe, C1, Stormberg, A4, Williams, P2, 1 Univ Georgia, Savannah River Ecology Lab, Aiken, SC, US2 Dept Env Health Sci, Univ Georgia, Athens, GA, US3 Dept Biol Sci, Univ South Carolina, Columbia, SC, US4 INEEL, Idaho Falls, ID, US
ABSTRACT- Direct assessment of the effects of potential mutagens is difficult and expensive with existing methodology. An ideal assay would be a sensitive and inexpensive animal model with a readily scored phenotype that responds quickly to exposure to bioavailable mutagens. To this end, we developed transgenic C. elegans, using the green fluorescent protein (GFP) reporter gene, incorporating a microsatellite construct as a molecular "On/Off" switch that responds to environmental mutagens. GFP produces a fluorescent product that is directly visible, eliminating the need for exogenous substrates and cofactors, and expensive or hazardous chromagens. GFP is visible in live animals, allowing repeated observations over time, and selection of mutants for breeding or further study. Detection of heritable mutations requires examination of offspring, and experiments with model mutagens in aquatic media resulted in high percentages of offspring expressing GFP. Future work will focus on the use of 96-well plates and flow cytometry to rapidly measure expression in large numbers of worms. This approach will greatly decrease the cost and effort required to assay mutagenicity. Additionally, this detection system uses an animal whose genome has been sequenced completely and is an increasingly important model for biomedical research.
Key words: mutation assay, transgenic, nematode, microsatellite