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Quantitative PCR Analysis of Neurodevelopmental Gene Expression in c. elegans Quantitative PCR is a technique that can be used to analyze expression of specific genes at different developmental stages or in response to changing environmental conditions. My goal for this project was to establish a working protocol for quantitative PCR so that it can be implemented in future C. elegans research and be developed as a lab exercise for Biology classes taught at Occidental College. A qPCR protocol was established, and we are currently examining the expression of the tba-1 gene and several sod (superoxide dismutase) genes in C. elegans. tba-1 encodes an alpha-tubulin that plays a role in the development of motor neuron synapses and axons in C. elegans. The dominant tba-1(ju89) mutation causes defects in locomotion and neuronal development. tba-1 expression in 5 different strains was analyzed: N2 (wild type), tba-1(ju89) , and three tba-1 suppressor strains. Superoxide dismutase is an enzyme that catalytically removes the O2- radical and protects the organism from oxidative damage during aging. Mutation in SOD1 is known to be one cause of motor neuron death in Lou Gehrig’s disease. In C. elegans, five sod genes are known to regulate life span, reaction to stress, dauer formation, and metabolism and have distinct developmental expression patterns. Quantitative PCR analysis of sod gene expression in wild type, dauer animals and several mutant strains is in progress. Support provided by: Mary S. Caswell Endowment |
