Diversity Analysis Of Macaca Mulatta MHC DRB Alelles Exon 2 From Sichuan

76 indivalduals from 5 Sichuan Macaca mulatta wild groups have been selected to uesed for DRB gene exon 2 partial sequences diversity analysis by PCR amplitication, DGGE seperation and gene colone.In all 76 Macaca mulatta samples,the gene typles was listed for each sample,meanwhile,new allelic gene was named.After all amimo acid sequences was analysised,applied N-J methouds to constract DRB gene phylogenetic trees that contant the other specis primates genes which were downloaded from NCBI gene bank. Finally,in 5 groups,gene evolution pressurs,inter-group gene average genetic distance, between-group genetic distance,each one was anlysised.The results for each analysis were listed:1) 69 alleles has been detected by PCR amplification,DGGE and gene colone.57 of the 69 alleles are newly.In 5 populations,DRB1,3,4,5 and W,the 5 local alleles has been detected,meanwhile,21 sub-locus have been observed.Each sub-locus show the populational perticularies:Sub-locus DRB*W25 and DRB*W37 the two sub-locus only were detected in population Xiaojin;Sub-locus DRB*W31 and DRB*W40 have been only detected in population Heishui.The other sub-loci,have been observed in more than 2 populations simultaneously.2) After all gene sequences be translated into amimo acid ensued with sequence-blast by solftware MEGA4.0,the difference between locus amimo acids focus on the secondary constractureβregion,althrough,in the same locus,the distinction between two alleles focus onαregion.Compared with human equivalents the difference amimo acids comcentrate onαregions,aboutβregion the difference showed lower thanα.3) Gene phylogenetic tree by solftware MEGA4.0 was used by method N-J.From tree, consenqued that DRB*W07,DRB1*07,DRB*W02,DRB*W31 the 3 sub-locus did not detected in other primers except Macaca mulatta,may they belongs to Macaca mulatta perticulary,other loci can be finded in other primates yet.Locus DRB1*03,DRB1*10,DRB*W01 clusterd with human HLA-DRB1*13,DRB1*04 clusterd with human HLA-DRB1*04,DRB3*04 clusterd with human HLA-DRB*02,01,all the loci adhered with human counterparter may could be used for human immun-medicology.4) The average genetic distance in each group was caculated by solfteware MEGA4.0 with methods Nei-Gojibori-p and Kimular-2-parameters respectly.Results by method Kimular-2-parameters,the genetic distance in each group were arranged from high to low: group Hanyuan＞group Bazhong＞group Jiulong＞group Xiaojin＞group Heishui.This category document that,the average genetic distance influenced by uncletide acid disparity were the lowsted in group HeiShui and the hightest in group HanYuan.But results by method Nei-Gojibori-p,the genetic distance in each group were arranged from high to low were dismarch with results by method Kimura-2-parameter:group Xiaojin＞group Hanyuan＞group Jiulong＞group Heishui＞group Bazhong.This categry recover different with results by method Kimura-2-paramete,by method Nei-Gojibori-p,the average genetic distance influenced by amimo acid disparity were the lowest in group Bazhong and the highest in group Xiaojin.All the above conclude that:genetic distance influenced by nucletide acid disparity in inner-group,group Heishui was the lowest and group Hanyuan were the highest,however,by amimo acid disparity,group Xiaojin and Bazhong show the hightest and lowest respectily.The average genetic distance in 5 groups were caculated by sorlftware MEGA4.0 resoults show that the genetic distance between two groups dis-accordant with the real geographic setting.In DRB alleles evolution,although the dynamic in all populations were positive selections pressure,the pressure in each population show the differences that in population Xiaojin the pressure showed the hightest, but in Bazhong the pressure showed the lowest.