Med. Weter. 69 (3), 174-180, 2013

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Kosowska B., Strzała T., Brzezińska K., Grabowski K., Dobosz T., Lebioda A.
Molecular phylogenetic analysis of the domestic cat and the Eurasian lynx based on the Cyp21 gene
The aim of the study was to determine the nucleotide sequence of the CYP21 gene of the Eurasian lynx, eight representatives of five selected domestic cat breeds and a European mixed-breed cat on account of the key role of this gene product in adrenal steroidogenesis. Cats nuclear DNA was obtained from peripheral blood, while the lynx DNA was isolated from muscle tissue. 21HS gene amplification was performed in 10 parts with the unified PCR conditions. The reaction products were sequenced. On the basis of the determined nucleotide (nc) sequence of the gene, the sequence of amino acids (aa) of the protein was determined. The sequences obtained in our study and collected from the GeneBank were aligned using Mafft and subjected to phylogenetic analysis using the program MrBayes 3.2. A total of 11 SNP’s were detected in 9 tested cat breeds and an additional 3 indels in the mixed-breed cat. Between the studied cats and lynx a total of 33 SNP differences were determined. In addition, three indels were located in the lynx that were absent in cats. In domestic cats the 21HS gene occurs as a series of three long alleles: 2500bp, 2503bp, and 2504bp. Both alleles of lynx were 2502bp in length. Genetic distance was identified between the studied breeds of cats, as well as between cats and the lynx. A genetic analysis of Cyp21 nucleotide sequences of cats and lynx was performed against homologous sequences obtained from GeneBank, derived from 13 different vertebrate species. In addition, evolutionary distance was estimated between the amino acid sequences (aa) of the cat / lynx and homologous sequences obtained from the GeneBank and derived from 15 different vertebrate species. It was demonstrated that the aa sequence of cat is almost identical to the sequence of lynx. Overall, basing on 21HS aa sequence, the closest kinship links felines and canids, then felines with pigs and ruminants, after which were rodents. The lowest CYP21 kinship links aa sequences of felines with eels and birds, followed by humans and apes.
Key words: Felis catus, Lynx lynx, , Cyp21, CYP21, genetic analysis, phylogenetic analysis.