论文黑麂Ⅱ类MHC DQA基因第二外显子扩增的序列分析).docVIP

黑麂Ⅱ类MHC DQA基因第二外显子扩增及序列分析 侯翠兰,生命科学学院 摘 要：主要组织相容性复合体(major histocompatibility complex，MHC)基因在机体对病原体免疫方面的联系已经得到越来越多的理论支持，特别是其Ⅱ类的DQ, DR基因，其多态性水平在一定程度上衡量着种群的进化潜力。作为一种功能基因遗传标记，其在非模式物种自然种群中的运用越来越多。黑麂(Muntiacus crinifrons)为中国特有的濒危鹿科动物，其野生种群分布范围和数量正逐步缩小。本文扩增了51个黑麂样品的MHC-DQA 第二外显子部分序列，结果得到3个等位基因，彼此之间的核苷酸差异从7到20个（氨基酸差异从4到14个）。没有一个个体获得两个以上等位基因，这说明这些序列可能来自单座位。 抗原结合位点(antigen-binding site ABS）上的非同义替代（dN=0156±0.059）：麂鹿科MHC； DQA； 序列分析 The Study of MHCⅡ DQA exon 2 amplification and sequence analysis in the Muntiacus crinifrons Cuilan Hou, College of life science Abstract: The major histocompatibility complex (MHC) has gained the more and more theoretical evidences of the relevance in pathogen resistance. Particularly the DQ, DR genes, which the level of polymorphism indicated the potential evolutionary ability to some extent. As a genetic marker of functional gene, it used a lot in non-model species in natural populations. The distribution of the wild populations of Black Muntjac (Muntiacus crinifrons), an endangered species endemic to China, is very narrow and its number is decreasing. In this study, we amplified partial MHC class II DQA exon 2 sequences from M. crinifrons. As a result, three alleles were detected in 51 investigated samples, which differ from each other by sever to 20 nucleotide substitutions (four to 14 amino acid substitutions). No a single individual obtained more than two alleles, which suggests these sequences may be from a single locus. A significantly higher rate of non-synonymouse substitutions (dN=0．156±0.059) than synonymous substitutions (dS=0.012±0.011) in antigen-binding site (ABS) codons indicated positive selection in the DQA locus. Key words: Muntiacus crinifrons； Cervidae； MHC ； DQA；sequence analysis 引 言: 黑麂(Muntiacus crinifrons),属偶蹄目(Artiodactyla)鹿科(Cervidae)麂属(Muntiacus)，为我国特有动物，是典型的亚热带山地森林动物。在我国被列为国家一级保护动物，在濒危野生动植物物种国际贸易公约(CITES)和国际自然与自然资源保护联盟(IUCN)名录中分别被列入附录Ⅰ和易危(VU)级，被公认为目前世界上最珍稀的鹿科动物之一。黑麂现仅分布于皖南、浙西以及与之接壤的江西和福建的局部山区，野生种群数量十分稀少（盛和林，1987