Polymorphic Analysis Based On Mitochondrial DNA And MHC-DPB1 Gene In Different-altitude Area Of Rhesus Macaque

According to the geographical distribution,Rhesus macaque included six subspecies:Macaca mulatta vestita,Macaca mulatta mulatta,Macaca mulatta lasiotis,Macaca mulatta littoralis,Macaca mulatta brevicaudus,and Macaca mulatta tcheliensis in China.The organizational structure,physiological metabolism and immune tissue of Rhesus macaque be highly similar with human,so as the model animal in the human medical research.In this study,we uesd the techniques of conventional PCR and cloning sequencing from three aspectives to analysis the polymorphic of complete mitochondrial genome,mitochondrial DNA control region and MHC-DPB1 gene of Rhesus macaque species.Mainly including the following aspects of this work:1)The total length complete mitochondrial genome of Rhesus macaque subspecies were 16561 bp?16565bp and it included 13 protein-coding genes(PCG),2 Ribosomal RNA genes(rRNA)and 22 transfer genes(tRNA)and one non-coding region(CR).The mitochondrial genome structure was consistent with other vertebrates,a typical closed circular double stranded DNA molecule.All genes encod on heavy strand,expected the NADH6 gene and eight transfer genes encod on light strand.As the tRNASer(AGY)gene lost the D ring could not form the typical clover.The mitochondrial DNA control region total length were 1078?1087bp,and it mainly contained Central conserved domain,two Extended terminal associated sequence(ETAS I and ETAS2)and three Conserved sequence block(CSB1,CSB2 and CSB3).Based on the 13 PCGs to calculation the synonymous substitutions(Ks)and non-synonymous substitutions(Ka)and the ration of Ka/Ks.The result indicated that:The difference of Ka/Ks of NADH6 gene between Hainan subspecies and four subspecies was very significant(P<0.01),and the NADH6 gene evolved in Hainan subspecies accumulated relatively non-synonymous mutations.2)21 haplotypes were defined as 216 mitochondrial DNA CR sequences of Rhesus macaque(Xizang,Shi Baoshan,Kowloon,Qianling Mountain,South Bay Monkey Island and Wulongkou)group.The genetic diversity calculated based on haplotype:Tajima's D was tested at-2.533 and deviation from neutral mutation reached a significant level(P<0.05)of Wulongkou group,so it maybe experienced the historical group expansion.The genetic distance and genetic differentiation coefficient and gene flow of Rhesus macaque group showed a certain correlation with their distribution.Based on the haplotype of CR sequences to constructed the phylogenetic tree and mediation Network digram,the result showed that the haplotype of Rhesus macaque from different geographical location according to the origin were clusted together.3)26 effective alleles were defined by 372 Mamu-DPB1 gene exon 2 sequences of Rhesus macaque(Xizang,Shi Baoshan,Qianling Mountain,South Bay Monkey Island and Wulongkou)group and it included 17 alleles(new defined)and nine alleles had been reported.The Ka/Ks ratio less than 1 in all sites and Antigen-binding sites and Non-Antigen-binding sites of Mamu-DPBl genes exon 2 and had no significant difference(P>0.05),and it indicated that the Mamu-DPBlgene exon 2 of Rhesus macaque population had experienced a purely selective selection."Random-site" model detected Rhesus macaque groups Mamu-DPB1*21:02 allele had two positive selection sites(6 R,60 N)and the posterior probability less than 0.01.The Mamu-DPBl phylogenetic tree showed that:Cross-species polymorphism existed in Mamu-DPB1*01:17?Mamu-DPB1*06? Mamu-DPB1*01:11,12 new alleles alone gathered a branch.So we speculated that new alleles may be a new DPB1*allele pedigree in Rhesus macaque.