Laboratoire d'Immunogénétique moléculaire, EA 3034, Faculté de Médecine Purpan, Université Paul Sabatier, Toulouse 3, France.
Immunogenetics. 2011 Nov;63(11):703-13. doi: 10.1007/s00251-011-0550-8. Epub 2011 Jun 28.
We characterized the repertoire of MHC class I transcripts of a Malaysian cynomolgus macaque by the study of EST libraries derived from the mRNA extracted from six tissues (thymus, spleen, bone marrow, liver, heart and pancreas). The MHC class I transcripts present in a lymph node of the same animal were characterized by pyrosequencing of amplified cDNA fragments (515 bp from exon 2 to the beginning of exon 4). All pyrosequence consensus sequences, but three corresponding to rare transcripts, were identical to those obtained from EST libraries. In total, we characterized 19 classical class I transcripts in the Malaysian macaque studied here. By means of high-throughput sequencing of exon 2 amplified from the genomic DNA (190 bp), we characterized 38 classical class I genes in the genome of this animal. By comparison, using the same method, we found 23 classical class I genes in the genome of a MHC homozygous Mauritian animal (H2/H2). All these results suggested that the Malaysian animal was most probably heterozygous. This study reveals that the high-throughput pyrosequencing allows not only to characterize the MHC class I transcripts but also to estimate the number of MHC class I genes in the genome of cynomolgus macaque.
我们通过研究从马来西亚食蟹猴的六种组织（胸腺、脾、骨髓、肝、心和胰腺）提取的 mRNA 衍生的 EST 文库，对其 MHC I 类转录本的库进行了特征分析。通过对扩增 cDNA 片段（来自外显子 2 的 515 个碱基到外显子 4 起始处）的焦磷酸测序，对同一动物淋巴结中的 MHC I 类转录本进行了特征分析。所有焦磷酸测序的共识序列，但有三个对应于罕见转录本的序列除外，均与从 EST 文库中获得的序列相同。在本研究中，我们总共鉴定了马来西亚猕猴中 19 个经典的 I 类转录本。通过对从基因组 DNA 扩增的外显子 2 进行高通量测序（190 个碱基），我们鉴定了该动物基因组中的 38 个经典 I 类基因。相比之下，使用相同的方法，我们在 MHC 纯合毛里求斯动物（H2/H2）的基因组中发现了 23 个经典 I 类基因。所有这些结果表明，马来西亚动物很可能是杂合子。这项研究表明，高通量焦磷酸测序不仅可以对 MHC I 类转录本进行特征分析，还可以估计食蟹猴基因组中 MHC I 类基因的数量。
