Development of HPRT-Mutants and the Resultant c DNA by Metabolization of Benzo[a]pyrene-7,8-dihydrodiol
Czaiczynski, Brion Jacob
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The purpose of this study was to investigate the mutagenic effects of Benzo[a]pyrene-7,8-dihydrodiol (BPD). By using DNA repair deficient cells from a person diagnosed with xeroderma pigmentosum (XP), it was then possible to increase the mutagenic effects of a known toxicant. BPD was added to XP skin fibroblast cells that can actively metabolize this compound. In order to study the specific effects, hypoxanthine phosphoribosyltransferase (HPRT) was selected as the target gene. By variations in media, HPRT- cells were selected. These cells were used as a template for RT-PCR. Once the HPRT gene cDNA is produced from these cells, it will then possible to sequence this mutated gene and find regions of the gene that have a high mutation frequency. There was success in both creating HPRT- mutants and making of the subsequent cDNA, however the sequencing portion of this study has not yet been completed.