DMD免疫缺陷型小鼠模型的建立及其基因型鉴定

目的建立假肥大型肌营养不良症(DMD)免疫缺陷型小鼠(scid/mdx双基因缺陷型,scid-/-/mdx-/-),以及其抗肌萎缩蛋白基因(dystrophin)和scid基因的PCR鉴定方法。方法利用经典遗传学的杂交-互交方法,将非糖尿病-严重联合免疫缺陷(NOD-scid)雄鼠(scid-/-)和mdx雌鼠(mdx-/-)进行交配,提取杂合子F2代小鼠的基因组DNA,对dystrophin以及scid基因进行片段扩增,电泳后观察结果。结果46只杂合子F2代小鼠中共鉴定出13只dystrophin和scid双基因缺陷型的纯合小鼠,将人骨髓间充质细胞移植入小鼠,能整合分化成骨骼肌细胞。结论实验成功的得到了DMD免疫缺陷型小鼠,并提供了一种简单快速准确的PCR鉴定方法,为DMD的实验研究中的异种间移植提供了有效的动物模型。

Establishment of DMD immunodeficient mouse model and its gene identification

Objective To establish duchenne muscular dystrophy(DMD) immunodeficient mouse model and PCR methods for the identification of its dystrophin and scid genotypes. Methods Following the cross-backcross method,NOD-scid male mouse was mated with mdx female mouse.The genome DNA was extracted from the tails of the F2 generations and PCR method was employed to amplify the normal and aberrant fragments of dystrophin and scid gene.The results were observed after electrophoresis. Results Among the 46 individuals of F2 generation,there were 13 homozygotes deficient in double gene of scid and dystrophin.Transplantation of the mesenchymal stem cells into these mice,it was found that the cells could integrate with the mouse cells and even differentiation into skeletal myogenesis cells. Conclusion DMD immunodeficient mouse model was successfully established,which may facilitate the intensively research of DMD in xenogenic transplantation,and supplied a simple,rapid and precise PCR method to identify its genotype.