The Analysis Of Parts Of MHC Gene Frequencies And Polymorphism Of Macaca Mulatta Population In Yunnan

MHC gene diversities are widely existence in rhesus macaque(Macaca mulatta ) population from different geographical origin,it can influence the stability,reliability and reproducibility of the experimental result.Therefore,in order to evaluate the results of research,it's very important to understanding the immunogenetics background of our rhesus macaque population.Macaca mulatta MHC genetic characterization becomes a new researching hotspot.In this report,some locus located on Mamu classâ… andâ…¡gene of rhesus macaque were analyzed by polymerase chain reaction with sequence-specific primers and Single-Strand Conformation Polymorphism(SSCP)o Six alleles in MHC classâ… A region(from Mamu-A~* 01 to A~* 06 ) and ten alleles of four locus in MHC classâ…¡DRB region(Mamu-DRB1~*0303,030/06,0309,0403,1005/07;DRB3~*0403; DRB5~*0301/02;DRB6~*0101,0103,0104/05) of 140 Yunnan origin rhesus macaque were typed.92 of these rhesus macaque were from wild group of Yunnan JinDong,and 48 were from breeding colony in the primate center of our institute.The results showed that Mamu-A~*01 to A~*06 gene were missing in these research groups.For MHC classâ…¡DRB region,the results showed that the frequencies of DRB1 ~*0305/06,DRB1~*0309,DRB1 ~*0403,DRB3~*0403,DRB6~*0101,DRB6~*0103 in wild group was 34.8%,41.3%,27.2%,40.2%,40.2%,95.7%.On the other side the frequencies of these loci in breeding colony were 45.8%,58.3%,22.9%,66.7%, 54.2%,62.5%.The detection result of the rest loci were negative.Compared with other regions of Chinese rhesus macaque and Indian origin rhesus macaque(Indian group),Yunnan rhesus macaque shows a distinct difference of MHC gene.In addition, the differences of allelic polymorphism are not significant between wild group and breeding colony.In order to verify further correctness of amplification products of each allele and locus,the Mamu-DRB1~*0305/06 and 0309 alleles of classâ…¡gene were chosen to direct sequencing and SSCP detection.Two of three random sampling sequencing results of Mamu-DRB1~*0305/06 is completely identical with the reference sequence of GenBank,but another is mutated in 5 base pairs.The sampling sequencing result of Mamu-DRB1~*0309 is completely identical with the reference sequence of GenBank. The result of SSCP showes that more than 80%positive samples are the same with the sequencing samples,the primers have good specificity.Combind the method of sequencing with SSCP ensures the correctness of the result,avoid large-scale sequencing,and reduce the research cost.