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The dominant MHC class I gene is adjacent to the polymorphic TAP2 gene in the duck, Anas platyrhynchos

机译:显性的MHC I类基因与鸭(Anas platyrhynchos)中的多态TAP2基因相邻

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We are investigating the expression and linkage of major histocompatibility complex (MHC) class I genes in the duck (Anas platyrhynchos) with a view toward understanding the susceptibility of ducks to two medically important viruses: influenza A and hepatitis B. In mammals, there are multiple MHC class I loci, and alleles at a locus are polymorphic and co-dominantly expressed. In contrast, in lower vertebrates the expression of one locus predominates. Southern-blot analysis and amplification of genomic sequences suggested that ducks have at least four loci encoding MHC class I. To identify expressed MHC genes, we constructed an unamplified cDNA library from the spleen of a single duck and screened for MHC class I. We sequenced 44 positive clones and identified four MHC class I sequences, each sharing approximately 85% nucleotide identity. Allele-specific oligonucleotide hybridization to a Northern blot indicated that only two of these sequences were abundantly expressed. In chickens, the dominantly expressed MHC class I gene lies adjacent to the transporter of antigen processing (TAP2) gene. To investigate whether this organization is also found in ducks, we cloned the gene encoding TAP2 from the cDNA library. PCR amplification from genomic DNA allowed us to determine that the dominantly expressed MHC class I gene was adjacent to TAP2. Furthermore, we amplified two alleles of the TAP2 gene from this duck that have significant and clustered amino acid differences that may influence the peptides transported. This organization has implications for the ability of ducks to eliminate viral pathogens.
机译:我们正在研究鸭子(Anas platyrhynchos)中主要组织相容性复合体(MHC)I类基因的表达和连接,以期了解鸭子对两种医学上重要的病毒:甲型流感和乙型肝炎的易感性。在哺乳动物中,多个MHC I类位点,并且一个位点的等位基因是多态的,并共同表达。相反,在低等脊椎动物中,一个基因座的表达占主导。 Southern印迹分析和基因组序列扩增表明,鸭至少具有四个编码I类MHC的基因座。为了鉴定表达的MHC基因,我们从一只鸭的脾脏中构建了未扩增的cDNA文库,并筛选了I类MHC。 44个阳性克隆,鉴定出4个I类MHC序列,每个序列具有大约85%的核苷酸同一性。等位基因特异性寡核苷酸与Northern杂交的杂交表明,这些序列中只有两个被大量表达。在鸡中,主要表达的MHC I类基因位于抗原加工(TAP2)基因的转运蛋白附近。为了研究是否在鸭子中也发现了这种组织,我们从cDNA文库中克隆了编码TAP2的基因。从基因组DNA的PCR扩增使我们能够确定主要表达的MHC I类基因与TAP2相邻。此外,我们从这只鸭中扩增了TAP2基因的两个等位基因,它们具有明显且成簇的氨基酸差异,可能影响所运输的肽。该组织对鸭子消除病毒病原体的能力有影响。

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