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中文摘要：
主要组织相容性复合体（Major histocompatibility complex，MHC）在脊椎动物的免疫系统中起着重要的作用，常作为适应性遗传标记应用于保护遗传学研究。长江江豚（Neophocaena phocaenoides asiaeorientalis）是惟一生活于淡水环境中的江豚种群，且已处于濒危状况。为了开发适用于长江江豚保护遗传学研究的MHC遗传标记，首次采用北象海豹（Mirounga angustirostris）的一对DRB基因引物对长江江豚的基因组进行扩增，从5个个体中成功扩增并测序得到5条MHC DRB基因第二外显子188bp的核苷酸序列。BLAST结果表明这5条DRB特异序列与Gen-Bank中自鲸（Delphinapterus leucas）的DRB2序列具有较高的同源性，从而证实得到了预期扩增位点。进一步分析发现：这5条序列在4个核苷酸位点上产生替代，翻译后氨基酸序列在3个位点上发生替代；均具有完整的开放阅读框；且核苷酸的非同义替代率远远高于同义替代率；此外，从同一个体分离到两种以上不同DRB核苷酸序列，暗示着长江江豚在DRB座位上可能存在基因重复现象。初步分析结果表明长江江豚的DRB基因具有核苷酸多态性和氨基酸多态性及潜在功能性，并经受着强烈的自然选择。因此，该DRB座位可以作为适应性遗传标记进一步用于长江江豚遗传多样性以及种群适应能力评估等保护遗传学研究。
英文摘要：
The major histocompatibilihy complex （MHC） is a large gene family encoding a set of trans-membrane proteins that play a key role to initiate immune responses in the vertebrates. DRB gene is one member of the MHC gene family and usually with high diversity, thus, it is prevalently used as an adaptive genetic marker in conservation genetics study. The endangered Yangtze finless porpoise （Neophocaena phocaenoides asiaeorientalis） is also known as the only freshwater finless porpoise population in Chinese waters. In order to develop MHC genetic marker that suitable for conservation genetics study, a primer pair that originally designed for DRB gene of North elephant seal （Mirounga angustirostris） was used to amplify the genome of the Yangtze finless porpoise. As a result, 5 DRB exon 2 unique sequences of 188bp （ including partial peptide binding region, PBR） were first isolated from 5 individuals via PCR and sequencing. When BLAST in Gen- Bank, these sequences showed high similarity （97%） with the DRB2 allele of the beluga （Delphinapterus leucas） , indicating the expected DRB loci were successfully obtained for the Yangtze finless porpoise. In further sequence analysis, nucleotide substitutions were decided at 4 sites with inducing amino acid substitutions at 3 sites, all of the 5 DRB unique sequences showing entire open reading frames, and also much more non-synonymous substitutions than synonymous substitutions were detected especially at PBR. Additionally, more than two unique sequences were identified from one individual, suggesting possible gene duplication at this DRB locus, All of the above results indicated that DRB gene in the Yangtze finless porpoise had potential function and high variability, and also underwent strong positive selection. Thus, the DRB locus isolated and identified in this study could serve as useful adaptive genetic marker in the conservation genetics studies of the Yangtze finless porpoise, such as MHC genetic diversity and population adaptability estimation.
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