Integration and expression of HLA-B2704 gene in transgenic mice

Objective To study the integration and expression of HLA-B2704 gene in HLA-B2704 transgenic mice. Methods HLA-B2704 gene was introduced into fertilized eggs of C57BL/6×Kunming and Kunming×Kunming by microinjection. The founder mice were screened by PCR for integration of HLA-B2704 transgene and were then further confirmed by southern blot. RT-PCR and flow cytometry were carried out to detect the expression of HLA-B2704 gene at mRNA and protein level. Results Ten F0 hybrid mice carried HLA-B2704 gene in 101 F0 hybrid mice, 3 of 64 (4.7%) hybrid mice from Kunming×Kunming and 7 of 37 (18.9%) hybrid mice from C57BL/6×Kunming background carried HLA-B2704 gene. Statistical analysis showed that there was significant difference in the integration rate between the two groups (P0.05). HLA-B2704 gene was expressed at mRNA level in the skin, colon, testicle and spleen tissue of the transgenic mice. The expression of HLA-B2704 gene on the cell membrane of their peripheral blood lymphocytes was 7.87% in HLA-B2704 transgenic mice at protein level. Conclusion Although the survival rate of fertilized eggs injected is not significantly different, the fertilized eggs of hybrid mice with C57BL/6 mice gene background are more easily integrated with exogenous gene, while the fertilized eggs of hybrid mice with Kunming mice gene background are integrated with difficult. Compared with Kunming×Kunming F0, C57BL/6×Kunming F0 is more suitable for production of transgenic mice. The complex of HLA-B27 heavy chain molecule combined with mouse β2 microglobulin is unstable, so it is difficult to express on the surface of the cell membrane.