纯合子NEOr转基因小鼠品系的建立及鉴定

目的　建立清洁级Neor 转基因小鼠纯合子品系。方法　通过胚胎移植生物净化方法获得 1 0只清洁级NeorF1 小鼠 ,按孟德尔遗传法则交配 ,用PCR、Southernblot杂交和交配实验检测相结合的方法筛选纯合子。结果　选育出 4只纯合子 ,并建系。该纯合转基因小鼠与野生型小鼠交配制备的胎儿成纤维细胞具有G4 1 8抗性 ,可作为ES细胞基因打靶培养中的饲养层细胞。结论　通过微生物学和遗传学上对Neor 转基因小鼠进行质量控制 ,使Neor 转基因小鼠达到了清洁级 ,并建立纯合子品系。

Founding and Identification of Homozygote NEOr Trangenic Mouse Strain

Objective To establish clean homozygote Neo r trangenic mouse strain.Methods Ten clean offsprings of Neo r transgenic mice free of 10 microorganisms infection were obtained through bio cleaning of embryo transfer. Among them five mice were identified as Neo r positive by PCR and Southern blot. The positive mice were bred by brother sister breeding methods to produce homozygotes. Results Four homozygote Neo r transgenic mice were selected through PCR, Southern blot and breeding experiment to form Neo r transgenic mouse strain. The mouse embryonic fibroblast (MEF) prepared from fetuses of female mouse mated with male Neo r transgenic mouse showed resistance against G418 as high as 1 mg/ml in the culture medium. This Neo r MEF can be used as feed cell layer when ES cell gene targeting is conducted. Conclusion The above results show that we obtained clean animals through embryo transfer, a genetical homozygote Neo r transgenic mouse strain.