Identification of novel polymorphisms and two distinct haplotype structures in dog leukocyte antigen class I genes: DLA-88, DLA-12 and DLA-64
The current information on the polymorphism variation and haplotype structure of the domestic dog leukocyte antigen (DLA) genes is limited in comparison to other experimental animals. In this paper, to better elucidate the degree and types of polymorphisms and genetic differences for DLA-88, DLA-12 and DLA-64, we genotyped four families of 38 beagles and another 404 unrelated dogs representing 49 breeds by RT-PCR based Sanger sequencing. We also sequenced and analyzed the genomic organization of the DLA-88 and DLA-12 gene segments to better define these two-gene DLA haplotypes more precisely. We identified 45 alleles for DLA-88, 15 for DLA-12 and six for DLA-64, of which 20, 14 and six, respectively, were newly described alleles. Therefore, this study shows that the DLA-12 and DLA-64 loci are far more polymorphic than previously reported. Phylogenetic analysis strongly supported that the DLA-88, DLA-12 and DLA-64 alleles were independently generated after the original divergence of the DLA-79 alleles. Two distinct DLA-88 and DLA-12 haplotype structures, tentatively named DLA-88–DLA-12 and DLA-88–DLA-88L, were identified, and the novel haplotype DLA-88–DLA-88L contributed to 32.7% of the unrelated dogs. Quantitative real-time PCR analysis showed that the gene expression levels of DLA-88L and DLA-88 were similar, and that the gene expression level of DLA-12 was significantly lower. In addition, haplotype frequency estimations using frequently occurring alleles revealed 45 different DLA-class I haplotypes (88-88L/12-64) overall, and 22 different DLA-class I haplotypes in homozygous dogs for 18 breeds and mongrels.
KeywordsDog Major histocompatibility complex; MHC Dog leukocyte antigen; DLA Polymorphism Haplotype structure Gene expression
We thank Toshihiro Watari in Animal Medical Center (ANMEC) at Nihon University, Shinichi Namba in Marble Veterinary Medical center (Fujisawa, Japan) and Oriental Yeast Co. Ltd. (Tokyo, Japan) for providing the dog blood samples. We also thank Atsuko Shigenari and Sayaka Ito in Department of Molecular Life Science in Tokai University School of Medicine for technical assistance. This work was supported by Nihon University President’s Grant for Multidisciplinary Studies.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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