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2002
LSU-HHMI Summer Undergraduate Research Program |
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 Jaime
Rueda (University of Texas – Pan American) (Vince L. Wilson,
LSU Dept. of Biological Sciences and Institute of Environmental
Studies) A novel assay used to detect germline
intragenic mutations caused by ENU in Drosophila melanogaster
In the field of genetics and germline mutagenesis there are
several assays that can detect point mutations but most lack
the desired sensitivity level required to detect one mutation
in thousands to millions of cells. This novel assay involves
combining polymerase chain reaction(PCR) with a restriction
enzyme digestion and ligase chain reaction. The overall process
is called PCR/RE/LCR, which can detect one mutant cell out of
a million wild type cells. The advantage of this assay over
others is its ability to detect a mutation at a desired sensitivity
in small amounts of DNA. In Drosophila melanogaster sperm exposed
to varying concentrations of a known mutagen, ethylnitrosourea(ENU),
we were able to detect one mutant cell out of 10000 wild type
cells. This result corresponds to those expected from the Sex
Linked Recessive Lethal(SLRL) test that is the standard in many
biological laboratories. This assay is not only useful for germline
mutagenesis but it can also provide a means to detect mutations
in oncogenic loci. Perhaps in the future this knowledge will
allow oncologists or geneticists to detect an oncogenic mutation
before it becomes a tumor so that treatment can be performed
much sooner and more efficiently.
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