骨髓特异性高表达neuritin转基因小鼠的构建

Establishment of transgenic mice model with overexpression of neuritin in bone marrow

目的构建骨髓特异性高表达neuritin转基因小鼠并进行繁殖、基因型鉴定,为进一步研究neuritin蛋白治疗外周神经病变奠定基础。方法将2对转基因小鼠loxp-stop-loxp-neuritin与转基因小鼠lyz-Cre/+进行饲养和繁殖,对其子鼠提取尾组织DNA,采用PCR方法鉴定小鼠基因型,获取基因型neuritinloxp/+_lyz-Cre/+即为骨髓特异性高表达neuritin小鼠。选择C57BL/6j野生型小鼠作为对照,并通过组织免疫荧光检测小鼠骨髓neuritin的表达量,进一步验证neuritin loxp/+_lyz-Cre/+小鼠的准确性。结果 2种转基因纯合子小鼠均有繁殖能力,相互杂交其子鼠基因型为neuritin loxp/+_lyz-Cre/+、neuritin loxp/-_lyz-Cre/+、neuritin loxp/+_lyz-Cre/-、neuritin loxp/-_lyz-Cre/-,其繁殖结果基本符合孟德尔遗传定律。免疫荧光结果显示:neuritin loxp/+_lyz-Cre/+小鼠骨髓neuritin表达量显著高于野生型小鼠(P<0.05)。结论 PCR是小鼠基因型鉴定的可靠方法。雄性lyz-Cre/+小鼠与雌性neuritin loxp/+小鼠交配是获得neuritin loxp/+_lyz-Cre/+小鼠的有效途径。

Objective To establish transgenic mice model with over expression of neuritin in bone mar-row,for the further study on the function of neuritin protein in the treatment of peripheral neuropathy. Methods Two pairs of transgenic mice（loxp-stop-loxp-neuritin and lyz-Cre/＋）were fed and propagated,the DNA from themice tails extracted and the genotype of transgenic mice identified by PCR. The wild type mice with B6 were as-signed as the controls,and the immunofluorescence was used to detect the accuracy of the neuritinloxp/＋_lyz-Cre/＋. Results The two trensgenetic homozygous mice had the ability to reproduce,and the hybrid offspringswere neuritinloxp/＋_lyz-Cre/＋,neuritinloxp/-_lyz-Cre/＋,neuritinloxp/＋_lyz-Cre/-,neuritinloxp/-_lyz-Cre/-. The re-sults were met with the Mendel′s law. The results of immunofluorescence showed that the expression of neuritin ofneuritinloxp/＋_lyz-Cre/＋ mice in bone marrow was significantly higher than the wild type mice（P0.05）. Conclusion The PCR method is of high reliability for identification of sub pus genotype and the female neuritinloxp/＋mice mating with the male lyz-Cre/＋ ones is an effective way for obtaining the neuritinloxp/＋_lyz-Cre/＋ mice.