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Development of a robust genetic test for hyperkalemic periodic paralysis (HYPP) in quarter horses
Author(s) -
Traver Samantha,
Horton Stephen
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.863.2
Subject(s) - mutant , primer (cosmetics) , genetics , biology , polymerase chain reaction , microbiology and biotechnology , gene , chemistry , organic chemistry
A single nucleotide substitution from cysteine to guanine 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). Genomic DNA from both whole blood and saliva swab samples were isolated from horses with wild type phenotypes for HYPP. As expected for an unaffected individual, our test showed the expected PCR product using the wild type specific primer pair, while the mutant specific primer pair did not generate a product. We then varied annealing temperatures and magnesium concentrations to optimize the PCR and found a temperature of 60°C and 3 mM magnesium to be optimal. We next plan to obtain DNA from both carrier horses and individuals expressing the full‐blown mutant phenotype. DNA isolated from HYPP animals should produce a PCR product with only the mutant specific primer pairs, and carriers for HYPP should produce a product in both wild type and mutant specific PCR reactions.