Evolution Analysis Of Chinese Macaques And The Study Of Special Genetic Marker

When identifying the doubtful samples of macaques obtained in wild survey and protecting work,it was difficult to identify these which were usually polluted,putrescent,part and denier(for example:hair,bones, nail,viscera,muscle fibre and so on).It was very significative to found a sensitive,accurate,dependable method which can identify 5 macaques.Test 1:According to both side sequences of cytb gene of human and Macaca mulatta,we designed and compose a pair of primers:Ca,Cs,and amplified cytb gene of two macaques(Macaca nemestrina and Macaca assamensis).1140 bp of cytb genes was obtained by nucleotide sequencing (GenBank submission number of Macaca nemestrina:EU204975).Used DNAstar software to compare these sequences,seted up phylogenetic tree and carried out phylogenetic analysis.This contributes to molecule identification of these macaques.Test 2:First,the gene sequences of cytb downloaded from the Genbank, which came from seven common species,five macaques and ten sib species, were alined by the DNAstar software,and looked for conservative mutational site of five macaques.Then according to the basic principle of primer designing,designed and synthesized a pair of specific macaque primers:Ms and Ma.The total DNA were extracted from muscles,hairs and feces of ten near original species and seven common species,then cytb were amplified to examine the specificity,sensitivity and repeatability of Ms and Ma.The result showed that Ma and Ms particularly amplified the DNA of five macaques,while didn't amplify the common species and near original species,and the rate is 100%.It showed this method is stable, sensitive,and reliable.So that obtained a method with which the macaques can be identified,and established a molecule biology method of identifying five macaques in the lab.It was a convenient,reliable method for identification of partial and little the macaque samples.Looked for conservative mutation site by aligning cytb gene sequences of five macaques.According to this mutation site,using the Primer 5.0 and Oligo 6.0 software,designed specific primers of each macaque,and examined their specificity,sensitivity and repeatability by PCR.The optimized the conditions of PCR.The result showed that each macaque's specific primers can amplified the DNA of corresponding macaque.In other words,the primer is very particular,and it didn't amplify the other species except corresponding macaques,and the rate was 100%.The method was stable,sensitive and reliable,and can indentify the little sample of each corresponding macaques.So established a molecule biology method of identifying each macaque in the lab,and it provided a convenient, reliable method to identify the remains and partial macaque samples obtained from wild investigates.