Cx43基因条件敲除小鼠的繁殖、基因型鉴定和骨髓检测  

Breeding,genotyping and bone marrow examination of the conditional connexin 43 knock out mice

作　　者：徐燕霞[1] 周东明[1] 傅晋翔[1] 

机构地区：[1]苏州大学附属第二医院血液科,江苏省215004

出　　处：《江苏医药》2017年第11期753-756,共4页Jiangsu Medical Journal

基　　金：国家自然科学基金(81272631)

摘　　要：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 explore the breeding,genotyping and mechanism of the onset of myelodysplasia in the conditional connexin 43（Cx43）knock out mice（Cx43KO mice）.Methods Two pairs of transgenic mice（Cx43loxP/loxP and Lyz-Cre/＋）were inbreeded to produce Cx43loxP/-_Lyz-Cre/＋ mice,females of which were used to mate with the male Cx43loxP/loxP mice to finally get the Cx43loxP/loxP_Lyz-Cre/＋ mice（Cx43KO mice）.After genome DNA was extracted from the mice tails,the genotype was identified by PCR and agarose gel electrophoresis.The real-time PCR was used to detect the accuracy of the Cx43 KO mice.Results Both breeding and reproduction of Cx43 KO mice were successful.It was fruitful to obtain four genotype mice of Cx43loxP/loxP_Lyz-Cre/＋,Cx43loxP/-_Lyz-Cre/＋,Cx43loxP/loxP and Cx43loxP/-.The results of breeding were met with the Mendel’s law.The expressions of Cx43 mRNA in bone marrow,liver and spleen were lower in Cx43 KO mice than those in heterozygote ones（Cx43loxP/-_Lyz-Cre/＋）（P〈0.05）.Conclusion The PCR method is able to correctly identify the genotype of the offspring.Mating female Cx43loxP/-_Lyz-Cre/＋ mice with male Cx43loxP/loxP ones is a useful approach to obtain the Cx43 KO mice.

关 键 词：基因敲除 连接蛋白43 基因型鉴定 聚合酶链反应 

分 类 号：R965[医药卫生—药理学]