Establishment Of Immortal Cell Banks Of Ha-Li And Qi-Li And Research On The Y-DNA Polymorphism

The immortal cell banks of the Ha-Li and Qi-Li in Hainan Island were successfully established by Epstein-Barr virus transformation of B-lymphocytes and addition of cyclosporine to inhibit the activity of T lymphocytes. In the present study, 142 immortal cell lines of the Ha-li and the Qi-Li were established. The DNA banks of the Benti-Li, the Qi-Li and the Ha-Li were also set up.We used 10 Y chromosome markers to study affinities among 3 Li ethnicvpopulations (Benti-Li, Qi-Li and Ha-Li) and their paternal relations to other populations. The polymorphism survey included three bialleli sites (YAP; DYS199; M9), six Y chromosome specific STR polymorphic loci (DYS19; DYS3891; DYS390; DYS391; DYS393; DYS395),and a minisatellite locus. Analyzing the Y-DNA SNP sites, we found the C allele and the absence of YAP element were displayed in all 289 male individuals of 3 Li populations. M9 locus has G allele in all samples except one. These results suggest that 3 Li populations have common paternal origin at these SNP loci. The MSY2*3 was found at different frequency in three Li subgroups from Haina Island (1.12% in Benti-Li,7.22% in Qi-Li,19.63% in Ha-Li). Also we investigated the polymorphism of 6 Y-STR, 289 healthy male individuals in these 3 Li populations involved. The analysis of six Y-chromosome-specific microsatellite loci revealed a high level of polymorphism in three Li populations. Six of these loci were found to generate at least 67 different haplotypes and haplotypes diversity with the range of 0.886-0.975. The Ha-Li population revealed the highest level of genetic diversity as compare toIVBenti-Li and Qi-Li, while the Latter group was the most homogenous. Analysis of molecular variation (AMOVA) showed a moderate genetic differentiation among the 3 Li populations, which primarily result from the divergence between the Ha-li and the Qi-Li. These results might imply that Ha-Li genetic diversity is the recent admixture with the different immigrant populations. The conclusion is supported by the distribution of the most frequent allele (MFA) in Ha-Li according with Chinese Han in more Y-STR sites than Qi-Li and Benti-Li. A network was constructed by five Y-STR (DYS19, DYS3891, DYS390, DYS391, DYS393) haplotypes. In the network, no evident haplotype clustering was showed on the basis of different Li subgroup populations. Moreover, three Li populations Benti-Li, Qi-Li and Ha-Li are always clustered in most of the trees. The results suggest a close genetic relationship of these three populations. Some genetic evidences of origin and relationship among three Li populations have been shown in this research, which indicates reference populations for detecting male migration events and should be useful in population genetic and forensic application.' Similarly, the establishment of immortal cell lines and the DNA banks of the Ha-Li and Qi-Li not only systematically and permanently preserve the special Li ethnic group genome but also provide enough research materials for further studies.