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Utilizing Single Nucleotide Polymorphisms Microtubules are structural polymers essential to fundamental cellular mechanisms such as cell division, cell motility, and molecular transport within cells. Understanding the intra- and extra-molecular interactions of these dynamic polymers may help inform the treatment of neurodegenerative diseases characterized by breakdown of axonal transport. Previously, the C. elegans mutation tba-1(ju89) was characterized, and found to disrupt normal synapse formation and axonal outgrowth (Baran et al., 2010). Two independent isolates that suppress the tba-1(ju89) mutation, sup15 and sup33.1, were separately crossed into the polymorphic strain CB4856 to generate recombinant worms for genetic mapping. Determining the molecular identity of the suppressors will help reveal how microtubules are normally regulated in neurons and how disruption of these mechanisms can lead to synaptic defects. The Snip-SNP technique capitalizes on the naturally occurring differences, or polymorphisms, between the wild-type N2 strain and CB4856. This technique involves single nucleotide polymorphisms (snip-SNP’s) that can be detected by restriction enzyme digestion and subsequent gel electrophoresis. Preliminary data suggests that sup15 may be localized to the left arm of C. elegans chromosome III, while sup33.1 may contain a mutation in the middle of the X chromosome as well as a mutation on the left arm of chromosome III. Experiments are underway that employ other snip-SNPs to narrow down the locations of the two suppressors and to purify the suppressor DNA in preparation for whole-genome sequencing. Support provided by: National Science Foundation Grant to Prof. Baran |
