Yasukochi Yoshiki, Kurosaki Toshifumi, Yoneda Masaaki, Koike Hiroko
Department of Biosystems Science, the Graduate University for Advanced Studies, Hayama.
Genes Genet Syst. 2010 Apr;85(2):147-55. doi: 10.1266/ggs.85.147.
Genetic diversity estimation of the major histocompatibility complex (MHC) gene may be an important tool in the assessment of immune response ability against infectious disease. We were able to identify a near full-length expressed DQB sequence by RACE-PCR method from the Asiatic black bear, Ursus thibetanus in Japan. This is the first such full length expression in the Ursidae. The bear had at least one functional DQB locus. In phylogenetic tree analysis its DQB amino acid sequence formed a monophyletic group with DQB sequences from members of the order Carnivora and had a 90% nucleotide sequence similarity with the DQB allele of the California sea lion, Zalophus californianus. We compared the DQB amino acid composition of U. thibetanus with those of several other mammalian species including Homo sapiens. Amino acid residues known to be functionally important for human MHC genes, tended to be also conserved among other mammalian species while PBRs in the beta1 domain were heterogeneous among mammalian species. The DQB sequence obtained from the bear had not only no putative frameshifts or deletions but also no abnormal amino acid mutations such as had been observed in human DQB molecules. This suggests that the bear DQB sequence was an apparently functional DQB allele. As a preliminary study, we sequenced the exon 2 region of DQB alleles from genomic DNA, and succeeded to amplify the exon 2 of DQB loci. Our study will provide useful information for conservation genetics of the U. thibetanus as well as more generally regarding the mammalian MHC region.
主要组织相容性复合体(MHC)基因的遗传多样性估计可能是评估针对传染病免疫反应能力的重要工具。我们通过RACE-PCR方法从日本的亚洲黑熊(Ursus thibetanus)中鉴定出一个近乎全长的表达DQB序列。这是熊科中首次出现这样的全长表达。该熊至少有一个功能性DQB位点。在系统发育树分析中,其DQB氨基酸序列与食肉目成员的DQB序列形成一个单系群,并且与加利福尼亚海狮(Zalophus californianus)的DQB等位基因具有90%的核苷酸序列相似性。我们将西藏黑熊的DQB氨基酸组成与包括智人在内的其他几种哺乳动物的进行了比较。已知对人类MHC基因功能重要的氨基酸残基,在其他哺乳动物物种中也倾向于保守,而β1结构域中的PBRs在哺乳动物物种之间是异质的。从熊身上获得的DQB序列不仅没有推定的移码或缺失,也没有在人类DQB分子中观察到的异常氨基酸突变。这表明熊的DQB序列是一个明显功能性的DQB等位基因。作为一项初步研究,我们对基因组DNA中DQB等位基因的外显子2区域进行了测序,并成功扩增出DQB位点的外显子2。我们的研究将为西藏黑熊的保护遗传学以及更广泛的哺乳动物MHC区域提供有用信息。