hApoE7转基因小鼠纯合子的培育和鉴定

目的：载脂蛋白E(apoE)是血浆中参与脂质代谢的重要蛋白质，并且与迟发型早老性痴呆(AD)相关。为从整体水平研究人突变ApoE基因剂量与效应的关系，我们拟培育C57BL/6-hApoE7纯合子转基因小鼠。方法：将经Southern杂交鉴定为阳性的F1代小鼠进行互配，仔代经PCR初筛，再行Southern杂交鉴定，杂交结果经扫描定量分析以确定hApoE7基因整合的拷贝数，纯合子小鼠的拷贝数为杂合子的两倍。经此法鉴定的纯合子小鼠与正常C57 BL/6小鼠进行侧交以进行进一步的鉴定。结果：Southern杂交鉴定出F1代转基因小鼠6只，F2代转基因小鼠7只，其中纯合子3只。将所得到的纯合子小鼠与正常的C57 BL/6小鼠交配，仔代鼠经PCR鉴定结果全部为阳性。结论：通过培育C57 BL/6-hApoE7纯合子转基因小鼠，发现人ApoE7基因在转基因小鼠体内可以稳定遗传，转基因的遗传特征符合常染色体单位点整合。

Breeding and Identifying of the Homozygous hApoE7 Transgenic Mice

Objective Apolipoprotein E (apoE) is an important serum protein that plays a role in lipid metabolism. Moreover, apoE4 has been identified as a major risk factor for Alzheimer's disease (AD). In order to study the relationship between the gene dose and effect at the whole body level, we bred the homozygous hApoE7 transgenic mice. Methods F1 generation of the C 57BL/6-hApoE7 transgenic mice was mated, using PCR method and Southern blot to test the offspring.The image of the blot was done with laser scanning to identify the copy number of hApoE7, the copy number of the homozygote should be as twice as heterozygote. Then those homozygous mice were cross-bred with normal C 57BL/6 mice to test further. Results 6 F1 generation and 7 F2 generation transgenic mice including 3 homozygotes were identified by Southern blot. The offspring of the crossbreeding of the homozygous mice with normal C 57BL/6 mice were all positive by PCR examination. Conclusion Through breeding the homozygous hApoE7 transgenic mice, we found apoE7 gene could be inherited stably in the transgenic mice and the inheritance pattern of the transgene was compatible with a single autosomal integration site.