Using genetic sampling of hair to confirm origin and identify Florida Panthers

Using genetic sampling of hair to confirm origin and identify Florida Panthers

Alvin Sherman Library 4009

This project investigates the origin of a puma currently living in a zoo in Ecuador. Although this particular puma (named Sasha) is assumed to be of South American origin there is some speculation that she was brought to South America from Florida in the early 1980s. In order to determine Sasha’s most likely geographical origin and subspecies, DNA was extracted from hair from three known Florida panthers as well as from one of Sasha’s cubs. The target gene for this research is the mitochondrial NADH-5 (ND5) gene that is passed on from the maternal parent. Several variations (haplotypes) in this gene have previously been shown to be indicative of the different puma subspecies, which reflect geographical range. Following extraction and isolation of the DNA, primers for the mitochondrial ND5 gene were added and polymerase chain reaction (PCR) was performed in order to isolate this specific ND5 gene and amplify it. The PCR products were then run on a gel to confirm that the isolated section of DNA was 328 base pairs, the size of the ND5 gene. The restriction enzyme Bsma1 was then added to the gene. This restriction enzyme targets a single nucleotide difference between the North American and South American haplotype by cutting the ND5 gene into two similarly sized fragments and leaving the ND5 gene from the South American puma subspecies uncut. The differences in nucleotide sequence among puma subspecies (haplotypes) can also be investigated by sequencing the ND5 gene isolated from the different puma individuals.