Immunogenetics

, Volume 63, Issue 11, pp 703–713 | Cite as

Characterization of MHC class I transcripts of a Malaysian cynomolgus macaque by high-throughput pyrosequencing and EST libraries

  • Alice Aarnink
  • Pol-André Apoil
  • Ichiro Takahashi
  • Naoki Osada
  • Antoine Blancher
Original Paper

Abstract

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.

Keywords

MHC transcriptoma EST Pyrosequencing Macaca fascicularis 

Supplementary material

251_2011_550_MOESM1_ESM.doc (32 kb)
Supplementary Table 1Numbers of reads obtained by pyro-sequencing. (DOC 31 kb)
251_2011_550_MOESM2_ESM.doc (56 kb)
Supplementary Table 2Exon 2 of MHC class I genes obtained by blast in a single haplotypic sequence of Mamu class I region deposited by Shiina et al. (AB128049 accession genbank number). (DOC 56 kb)
251_2011_550_MOESM3_ESM.doc (51 kb)
Supplementary Table 3Characterization of MHC class II transcripts of a Malaysian macaque by EST libraries number of EST. The consensus sequences were obtained with 97 % identity in the full length coding region. (DOC 51 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Alice Aarnink
    • 1
  • Pol-André Apoil
    • 1
  • Ichiro Takahashi
    • 2
  • Naoki Osada
    • 3
  • Antoine Blancher
    • 1
  1. 1.Laboratoire d’Immunogénétique moléculaire, EA 3034, Faculté de Médecine PurpanUniversité Paul Sabatier, Toulouse 3, IFR150 (INSERM), CHU de ToulouseToulouse cedex 9France
  2. 2.Department of Biomedical ResourcesNational Institute of Biomedical InnovationIbarakiJapan
  3. 3.Division of Evolutionary GeneticsNational Institute of GeneticsMishimaJapan

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