Polymorphism Analysis Of MHC Class I A Sequences Of Cynomolgus Monkey

MHC (major histocompatibility complex) represents a tightly linked cluster of genes, which produce transmembrane glycoproteins on the cell surface, histo- compatibility antigen.The antigen by combining with antigen-binding site (antigen- binding site, ABS) can recognize the foreign peptide antigen, and present them to cytotoxic T cells and trigger the immune response to protect themselves from infection.Nonhuman primates are excellent models for studying human disease because they are immunologically similar to human and susceptible to many of the same pathogens. Rhesus macaques was previously widely used as an animal model in biomedicine study. Due to the export ban of India in 1978, the availability of rhesus macaques is restricted. More and more researchers have been using other species which is genetically close to rhesus macaques as a live animal model, such as cynomolgus macaques (macaca fascicularis). The value of cynomolgus macaques as a model for infectious disease research constantly improve. Cynomolgus macaques have previously been used in studies of Mycobacterium tuberculosis (Mtb), Ebola virus, and dengue virus.There is a finding that the variability of MHC significantly impact on the stability authenticity and repeatability of the experimental result. Considerable research has been conducted on the HLA and rhesus macaque (Macaca mulatta) MHC, but there are relatively limited studies on cynomolgus macaques MHC.This study targets at cynomolgus macaques immortalization A lymphocyte series and fresh blood of cynomolgus macaques. using the technology roadmap of―specificity primers PCR amplification-cloned-sequencing‖. Primers were designed in the conservative UTR, MHC around 3000bp were amplified .218 MHC class I alleles around 3000bp have been identified from 10 Macaca fascicularis. 11 of them , which have higher frequency , have been submitted to Genbank. BLAST analysis showed that the novel alleles include 7 Mafa-A alleles and 4 Mafa-AG alleles. Different Mafa-A alleles have been identified in each sample, it can be inferred that Mafa-A alleles have been duplicated and Mafa-AG alleles are derivative of Mafa-A alleles. Based on the new alleles, we analyze the alleles distribution among the samples and the phylogenetic tree. Eight exons of MHC were spliced through Genscan software and comparation with Rhesus monkey. The analysis of amino acid sequences translated by the new MHC gene sequences showed that exon 2 and exon3 have high polymorphism.Take GU215170 gene for example to study protein structure of the MHC I A loci of Cynomolgus macaques. Furthermore, their membrane region, signal peptide, secondary structure, tertiary structure were also predicted.