Isolation Of Microsatellite Markers In Tupaia Belangeri Chinensis And Analysis Of Groups Polymorphism

The tree shrews are classified into the order Scandentia and with a close affinity to primates. They have been proposed as a valid experimental animal in biomedical research. As the ones which researchers will use the most in the world, Tupaia belangeri chinensis are used to establish various disease models. However the experimental tree shrews are mostly the wild type, the genetic background is not clear, the genetic quality cannot be guaranteed, and the repeatability and scientificity of the results are poor. Therefore, control of genetic quality in experimental tree shrews is great necessity and scientific significance in the application of life science and biomedical research. But, there are few microsatellite loci reported in tree shrews. We are aim to find more new microsatellite loci and establish genetic control testing technology and standards in the artificial breeding tree shrew populations. Furthermore, we also analyze polymorphism analysis in the existing population groups. Firstly, we screen about 700 microsatellite loci from tree shrew genome sequence, then optimally selected about 100 loci to design primers, finally retained 33 pairs of primer. And adding 13 pairs of primer reported in references to the 33 pairs of primer for PCR amplification in Tupaia belangeri chinensis. According to the results of the agarose electrophoresis and polyacrylamide electrophoresis, we selected 22 microsatellite loci suitable for genetic testing. We randomly selected 15 males and 15 females from F1 generation of tree shrews as samples, a total of 30. Then the selected 22 microsatellite loci were used to do STR scan. The result shows:(1) 13 pair of primers have been chosen from the reference, including 7 pairs of primer of,3 pairs of which are available;3 pairs of primer of Tupaia minor,1 pair of which is available;3 Pairs of primer of Tupaia belangeri,2 pairs of which are available, all of these primers ultimately narrowed down to 6.The expected heterozygosity of TG4 among wild Tupaia glis is 0.76,well in the research results in 0.3;the expected heterozygosity of TG22 among wild Tupaia glis is 0.93,well in the research results in 0.37;the expected heterozygosity of JS188 among wild Tupaia longipeså'Œwild Tupaia tana are respectively 0.77 and 0.63, and in the research results in 0.56; the expected heterozygosity of SKTg22 among wild Tupaia longipeså'Œwild Tupaia tana are respectively 0.90 and 0.76, and in the research results in 0.53. the group STR scanning shows that among the 22 new loci, account of the weakness of some primers’ fluorescence signal.only 16 pairs had been chosen for analysis of groups polymorphism. The alleles of these 16 loci range from 2 to 15, mean alleles number is 6.375.Fragments size range from 123 to 432bp.The mean expected heterozygosity is 0.6138. Mean Shannon index is 1.265. Mean PIC is 0.563. Data analysis in our paper indicate that:(1) As microsatellite loci is not entirely in common use among different trees shrew species.it is necessary to search suitable microsatellite makers for Tupaia belangeri chinensis.(2)Compared to other tree shrews wild type populations, the genetic polymorphism in the population,our result is relatively low, which may be due to interspecific differences among different trees shrews species or a reasonable decline for the polymorphism of the first filial generation.(3)Based on polymorphism information content,the genetic diversity in the population we studied is highly polymorphic. The 16 new microsatellite loci we screen can provide high polymorphic information, which apply to genetic testing for Tupaia belangeri chinensis and give a scientific basis for tree shrew population’s genetic quality monitoring.