Identification of novel MHC class I sequences in pig-tailed macaques by amplicon pyrosequencing and full-length cDNA cloning and sequencing
Identification of novel MHC class I sequences in pig-tailed macaques by amplicon pyrosequencing and full-length cDNA cloning and sequencing
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DOI：
10.1007/s00251-009-0397-4
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发表时间：
2009-10-01
期刊：
影响因子：
3.2
通讯作者：
O'Connor, David H.
中科院分区：
文献类型：
--
作者：
O'Leary, Claire E.;Wiseman, Roger W.;O'Connor, David H.
猪尾猕猴(Macaca Nomestrina)在生物医学研究中提供了重要的动物模型，但由于对主要组织相容性复合体(MHC)I类基因的不完全了解，限制了该物种在HIV和其他疾病发病机制研究中的应用。在这里，我们描述了全面的MHC I类基因分型，使用焦磷酸测序来评估跨越MHC I类转录物高度多态的肽结合区的367个碱基的互补DNA(CDNA)-PCR扩增片段。我们检测到29个先前描述的MANE转录本，90个新的I类序列，以及8个共享的MHC IB类单倍型。我们使用这些基因分型数据来通知全长MHC I类基因的发现，表征了66个新的全长转录本。这些新的全长序列几乎是以前所描述的MANE-B序列数量的三倍。本文描述的全面的基因类型和全长MANE转录本为猪尾猕猴作为生物医学研究的模式生物增加了价值；此外，MHC基因分型和等位基因发现的协调方法可以扩展到其他特征不太好的非人类灵长类物种。
Pig-tailed macaques (Macaca nemestrina) provide important animal models in biomedical research, but utility of this species for HIV and other disease pathogenesis research is limited by incomplete knowledge of major histocompatibility complex (MHC) class I genetics. Here, we describe comprehensive MHC class I genotyping of 24 pig-tailed macaques, using pyrosequencing to evaluate a 367- bp complementary DNA (cDNA)-PCR amplicon spanning the highly polymorphic peptide-binding region of MHC class I transcripts. We detected 29 previously described Mane transcripts, 90 novel class I sequences, and eight shared MHC class IB haplotypes. We used this genotyping data to inform full-length MHC class I cDNA allele discovery, characterizing 66 novel full-length transcripts. These new full-length sequences nearly triple the number of Mane-B cDNA sequences previously characterized. The comprehensive genotypes and full-length Mane transcripts described herein add value to pig-tailed macaques as model organisms in biomedical research; furthermore, the coordinated method for MHC genotyping and allele discovery is extensible to other less well-characterized nonhuman primate species.