Chemical mutagenesis of unc-82 and other muscle-affecting genes in Caenorhabditis elegans
Duszynski, Robert J.
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Caenorhabditis elegans is a species of nematode that has been a model organism for scientific research since the 1970s (Brenner, 1973). The striated muscle cell within C. elegans is a highly ordered structure that contains actin and myosin filaments, anchoring structures, and regulatory proteins. For C. elegans, uncoordinated (unc) mutations cause abnormal development of muscle cells producing deformed structure and slow locomotion (Riddle et aI., 1997). The unc-82 gene encodes for a protein essential for myosin thick filament organization (Flanagan et aI., 2004). The unc-82 alleles e1323 and e1220 introduce a stop codon and missense mutation respectively, in the kinase domain (Hresko et aI., 1994). In this study, an ENU mutagenesis using an RW1596 strain of C. elegans was performed in an attempt to produce a mutation in the unc-82 kinase domain and subsequently one mutation (Ph3) in muscle organization was observed. Fluorescent light revealed clumping of myosin near the center of muscle cells in ph3 individuals. Attempts to run a complementation test with the unc-82 (e1323) allele were unsuccessful and the complementation of ph3 to unc-82 was undetermined. Attempts to map ph3 to a chromosome using mapping crosses were also unsuccessful. Genomic DNA from the phl allele on the unc-82 locus isolated from previous research was extracted. Gel electrophoresis confirmed that the kinase domain of unc-82 was amplified using PCR. Our results suggest that ph3 is an unknown allele that could either be a mutation on unc-82 with a novel phenotype or a mutation of an unknown gene. Future experiments will clarify the location of the phl and ph3 mutations as well as the role ofUNC-82 and other proteins in muscle development pathways of C. elegans.