title: Characterization of MHC class I transcripts of a Malaysian cynomolgus macaque by high-throughput pyrosequencing and EST libraries.
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.
authors.affiliation: Laboratoire d'Immunogénétique moléculaire, EA 3034, Faculté de Médecine Purpan, Université Paul Sabatier, Toulouse 3, France.
authors.forename: Alice
authors.initials: A
authors.lastname: Aarnink
authors.forename: Pol-André
authors.initials: PA
authors.lastname: Apoil
authors.forename: Ichiro
authors.initials: I
authors.lastname: Takahashi
authors.forename: Naoki
authors.initials: N
authors.lastname: Osada
authors.forename: Antoine
authors.initials: A
authors.lastname: Blancher
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000276:immunology / Q000378:metabolism; D005091:Exons; D005805:Genes, MHC Class I; D006579:Heterozygote; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology; D008099:Liver / Q000276:immunology / Q000378:metabolism; D008252:Macaca fascicularis / Q000235:genetics / Q000276:immunology*; D008297:Male; D008969:Molecular Sequence Data; D009206:Myocardium / Q000276:immunology / Q000378:metabolism; D010179:Pancreas / Q000276:immunology / Q000378:metabolism; D013154:Spleen / Q000276:immunology / Q000378:metabolism; D013950:Thymus Gland / Q000276:immunology / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D015395:Histocompatibility Antigens Class I