人白细胞抗原-B27(HLA-B27)转基因小鼠的建立

Construction of Transgenic Mice of Human Leukocyte Antigen B27(HLA B27)

人白细胞抗原 -B2 7( HLA-B2 7)与强直性脊柱炎 ( AS)有强相关性 ,为构建 AS动物模型以用于其发病机理和治疗的研究 ,将 HLA-B2 7基因与 β2 -微球蛋白基因等量混合后 ,利用显微注射技术注入昆明小鼠的1 3 70枚受精卵中 ,再将注射后存活的 1 0 50枚卵移植到 58只假孕母鼠体内 ,产仔 97只。PCR检测上述 97只转基因仔鼠 ,结果 2 7只为阳性。Southern blotting检测上述 2 7份 PCR阳性小鼠的基因组 ,结果 8只为阳性 ,表明已成功地建立了 HLA-B2

The transgenic mice of human leukocyte antigen B27 (HLA B27) has been generated by microinjection of the mixture of 6.5 kb HLA B27 gene and 15 kb β 2 microglobulin(β 2 m) gene into the fertilized eggs of mice. 1 050 out of 1 370 microinjected eggs have been transplanted to 58 pseudopregnant mice and 97 offspring were produced. Genomic DNA samples from the offspring have been detected by PCR and southern blotting techniques. The results showed that 27 out of 97 mice were positive by analysis of PCR and 8 mice were positive by southern blotting detection.