潮霉素抗性转基因BDF1小鼠模型的建立

目的建立具有潮霉素（hygromycin）抗性转基因BDF1小鼠，用于制备携有hygromycin抗性筛选标志ES阳性细胞克隆的饲养层。方法通过显微注射的方法，将含有潮霉素B磷酸转移酶基因片段（5.1kb）导入BDF1受精卵雄原核中，共注射169枚受精卵，然后将129枚受精卵细胞植入同期受孕的受体母鼠输卵管内。结果共产生37只转基因小鼠，经PCR和Southern检测获得9只阳性小鼠，对一只子代鼠进行RT-PCR检测证明hyg基因已经在肾、肌肉、脾内表达。结论成功的建立具有潮霉素抗性的BDF1转基因鼠，该模型动物可以为基因敲除研究提供良好的基础条件。

Establishment of a Hygromycin-Resistant Transgenic BDF1 Mouse Model

Objective To establish a hygromyein-resistant transgenie BDF1 model for preparation of feeder layers of hygromyein-resistant ES cells clones. Methods Hygremyein B phosphotransferase gene fragments were mleroinjected into the zygote male pronuelei of BDF1 mice. Altogether 169 zygotes were injected and 129 zygote cells were transplanted into the oviduct of aceeptor pregnant female mice. Results Genome DNA from mice tail were detected by PCR and Southern blot showing 9 of 37 were positives transgenie mice. One of the founders was analysed by RT-PCR and found that hyg was expressed in the kidney, muscle and spleen. Conclusion A hygromyein-resistant transgenie BDF1 mouse model has been successfully established. This model will provide favorable fundamental conditions for gene knock-out studies.