Amplification of Microsatellites of Japanese Raccoon Dogs (Nyctereutes procyonoides viverrinus) with Primers for the Dog ZUBECA4 Locus

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  • イヌZUBECA4プライマーによって増幅されるニホンタヌキマイクロサテライト座について

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The Japanese raccoon dog short tandem repeats (STRs, or microsatellites) were investigated using the primer set for the ZUBECA4 locus, one of the dog STR primers. The properties of Japanese raccoon dog PCR products were examined by electrophoresis and sequence analyses. Six alleles were detected by the electrophoresis analyses in nine Japanese raccoon dogs. The sequence analyses on the six alleles revealed that variants are based on the difference in the total number of repeats of GAAA and GCAA. The repeating units of Japanese raccoon dogs were simpler than those of dog alleles. The six alleles were designated as 12-16 and 18. Allele 15 and 18 were divided into 15^{12}, 15^{14}, 15^{15}, 18^{13} and 18^{14} sub-alleles by the times GAAA was repeated. Their frequencies were estimated to be: allele 12=0.10, allele 13=0.17, allele 14=0.17, allele 15^{12}=0.22, allele 15^{14}=0.06, allele 15^{15}=0.06, allele 16=0.06, allele 18^{13}=0.06 and allele 18^{14}=0.10. Heterozygosity was calculated to be 0.79 in STR alleles and 0.86 in sub-alleles. Further, new primers for Japanese raccoon dogs were developed based on the sequences of the Japanese raccoon dog alleles. This primer set was successfully amplified as a small-sized amplicon, which is more suitable for practical uses than dog primers. These findings indicated that the ZUBECA4 locus is a quite useful STR for species identification, individual identification, pedigree study and population study on Japanese raccoon dogs.

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