Date of Award

6-2011

Document Type

Open Access

Degree Name

Bachelor of Science

Department

Biology

First Advisor

John Horton

Language

English

Keywords

muscle tremors, weakness, DNA, equine, horse

Abstract

A single nucleotide substitution in a region of the skeletal muscle sodium channel gene (SCN4A) is known to cause an equine genetic disorder known as Hyperkalemic Periodic Paralysis (HYPP). The clinical effects of this disorder range from little or no symptoms to frequent episodes of muscle tremors, weakness, and/or complete collapse. Oligonucleotide primer pairs were designed for both the wild type and mutant alleles of the SCN4A gene for use in Amplification Refractory Mutation System-PCR (ARMS-PCR). These primers were tested with genomic DNA isolated from whole blood, saliva swabs, and hair of individual horses. It was determined that horse hair represented the most easily obtainable and reliable source for genomic DNA isolation. DNA was isolated from the hair of unaffected (wild-type) horses (N/N), a carrier for HYPP (N/H) and a homozygous mutant for the disease (H/H). As expected for the unaffected individual, a PCR product using the wild type-specific primer pair was generated, while the mutant-specific primer pair did not produce a product. DNA isolated from HYPP animals produced a PCR product with only the mutant-specific primer pair, and the carriers for HYPP produced a product in both wild type and mutant-specific PCR reactions. This robust DNA-based test was shown to generate an unambiguous assignment of the genetic status of the horses tested with respect to the HYPP trait.

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