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目的探讨连接蛋白43(Cx43)基因条件敲除小鼠(Cx43KO小鼠)的繁殖、基因型鉴定及其导致的骨髓增生异常机制。方法将引进的2对转基因小鼠Cx43loxP/loxP和Lyz-Cre/+进行交配饲养和繁殖,选取子一代雌性Cx43loxP/-_Lyz-Cre/+与雄性Cx43loxP/loxP合笼交配,获得Cx43loxP/loxP_Lyz-Cre/+小鼠(即Cx43KO小鼠)。提取鼠尾组织基因组DNA,采用PCR方法鉴定小鼠基因型,RT-PCR方法进一步验证Cx43KO小鼠的正确性。结果 Cx43KO小鼠繁殖成功,成功获得Cx43loxP/loxP_Lyz-Cre/+、Cx43loxP/-_Lyz-Cre/+、Cx43loxP/loxP、Cx43loxP/-四种基因型小鼠;其繁殖结果符合孟德尔遗传定律。Cx43KO小鼠骨髓、肝、脾Cx43mRNA的表达均较杂合型小鼠下降(P<0.05)。结论 PCR方法可准确鉴定子鼠的基因型。雌性Cx43loxP/-_Lyz-Cre/+小鼠与雄性Cx43loxP/loxP小鼠交配是获得Cx43KO小鼠的有效途径。
Objective To investigate the breeding, genotypic identification and the mechanism of myelodysplastic abnormalities in connexin 43 (Cx43) knockout mice (Cx43 KO mice). Methods Two pairs of transgenic mice, Cx43loxP / loxP and Lyz-Cre / +, were bred and bred. The first generation female Cx43loxP / -_ Lyz-Cre / + was crossed with male Cx43loxP / loxP to obtain Cx43loxP / loxP_Lyz- Cre / + mice (ie Cx43 KO mice). The genomic DNA of the tail tissue was extracted and the genotypes of the mice were identified by PCR. The correctness of Cx43KO mice was further verified by RT-PCR. Results Cx43KO mice were successfully propagated and successfully obtained Cx43loxP / loxP_Lyz-Cre / +, Cx43loxP / -_ Lyz-Cre / +, Cx43loxP / loxP, Cx43loxP / - mice. The reproductive results were in accordance with Mendel’s law of inheritance. The Cx43 mRNA expression in bone marrow, liver and spleen of Cx43KO mice was lower than that of heterozygous mice (P <0.05). Conclusion PCR method can accurately identify the genotypes of offspring. Mating female Cx43loxP / -_ Lyz-Cre / + mice with male Cx43loxP / loxP mice was an effective way to obtain Cx43 KO mice.