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Genetic Analysis of Classically and Embryologically Mutagenized C. Elegans
Chatley, Kevin Berg, Christian
Chatley, Kevin
Berg, Christian
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2013
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2013-05-03
Abstract
Both embryonic and classical mutagenesis of C. elegans were performed to analyze the difference in mutation rate and spectrum (i.e., types of mutations) observed upon genetic analysis using the chemical mutagen ethyl methane sulfonate (EMS). With embryonic mutagenesis, embryos were exposed to EMS at the one to four cell stage. With classical mutagenesis, the mutations are typically observed in the F2 generation; however, with embryonic mutagenesis, these mutations were seen in the F1 generation, indicating these mutations occurred early in development. For this project, 21 mutants were obtained, and 17 of these were analyzed and found containing mutations intragenic to unc-58 confirmed by previous genetic crosses. These strains are being reanalyzed from a previous, similar experiment to check for correctness and completeness, as a higher mutation rate was recorded than what was expected. In addition, 5 classically mutated strains were analyzed containing similar mutations. All of these strains were used for DNA extractions, polymerase chain reactions for the unc-58 sequence, and then genetically analyzed for mutations in the gene. Because of the difference in cell cycle checkpoints in embryos and adults, we expect to observe a difference in mutation rate between the two populations sequenced. From this study, we hope to gain scientifically significant knowledge with respect to the lack of homologous genes in human embryos.
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Biology