神经鞘磷脂合成酶2基因敲除小鼠的饲养繁殖和鉴定

目的:繁殖和鉴定神经鞘磷脂合成酶2(SMS2)基因敲除(-/-)小鼠。方法:将引进的SMS2杂合子(+/-)小鼠采用杂交、回交、互交的方法进行繁殖,一旦获得雄性和雌性SMS2纯合子(-/-)小鼠,便按照同样方法进行繁殖;用酚氯仿提取法提取小鼠基因组DNA,PCR扩增,琼脂糖凝胶电泳对小鼠基因型做出鉴定;将SMS2野生型(+/+)小鼠的产仔数作为正常对照组,与SMS2(+/-)和SMS2(-/-)小鼠的产仔数进行比较,用单因素方差分析、Dunnett检验处理数据。结果:SMS2(-/-)小鼠的饲养繁殖鉴定获得成功,单因素方差分析SMS2(+/+)小鼠、SMS2(+/-)小鼠以及SMS2(-/-)小鼠三组产仔数比较无显著性差异(P>0.05),Dunnett检验SMS2(+/-)小鼠与正常组比较产仔数无显著性差异(P>0.05),SMS2(-/-)小鼠与正常组比较产仔数无显著性差异(P>0.05)。结论:用正确的繁殖方法和鉴定方法可以成功获得SMS2(-/-)小鼠。

Breeding, reproduing and identifying SMS2 gene knockout mice

Objective: To reproduce and identify SMS_2 gene knockout(-/-)mice. Methods: SMS_2 heterozygote mice was reproduced with crossbreed,backcross and intercross methods.Male homozygote mice were selected to mate with the female homozygote mice for acquiring them with the same methods.Hydroxybenzene and chloroform were used to pick up the mice genome DNA,PCR was used to enlarge,gelose gel electrophoresis was used to identify.The reproducing rate of SMS_2(+/+) mice was used to be the control group,camparing with the reproducing rate of SMS_2(+/-) mice and SMS_2(-/-) mice,the date was dealt with ANOVA and Dunnett test. Results: Breeding,reproducing and identifying of SMS_2(-/-) mice were successful,ANOVA showed that the reproducing rate compared with SMS_2(+/+)mice,SMS_2(+/-)mice and SMS_2(-/-) mice were not significantly difference,Dunnett test showed that the reproducing rate compared with the control group and SMS_2(+/-) mice were not significantly difference,the reproducing rate compared with the control group and SMS_2(-/-) mice were not significantly difference as well. Conclusion: Appropriate methods for reproducing and identifying are the effective way for acquiring SMS_2 knockout mice.