TNNI2突变转基因小鼠模型的建立

Construction of TNNI2 Mutation Transgenic Mice

目的建立TNNI2突变转基因小鼠模型。方法构建pEGFP-tnni2转基因构件,TNNI2基因的第175个氨基酸缺失,通过原核显微注射法,把线性化、纯化后的外源基因pEGFP-tnni2注射入BDF1小鼠受精卵中,胚胎移植给同期发情的假孕受体母鼠,获得子代小鼠。用PCR和Southern方法检测子代鼠尾DNA鉴定基因型。通过RT-PCR方法检测tnni2基因表达。结果移植注射胚胎115枚给4只假孕小鼠共出生了23只后代鼠,经PCR和Southern方法检测得到4只阳性小鼠。对其子代进行RT-PCR检测,tnni2基因在肌肉、心脏内表达。结论通过显微注射法使外源基因pEGFP-tnni2(TNNI2基因的第175个氨基酸缺失)在小鼠基因组中得到整合,建立了转pEGFP-tnni2的转基因小鼠模型。

Objective To construct the animal model of TNNI2 mutation transgenic mice. Methods pEGFP-tnni2 （p. K175del） was constructed, linearized and purified, then microinjected into fertilized mouse eggs. These eggs were transplanted into the pseudopregant mice. The genotype of transgenic founders were identified by PCR and Southern blot. The expressions of tnni2 transcript in the tissues of the transgenic mice were detected by RT-PCR. Result Transgenic BDF1 mice were obtained by microinjection. PCR and Southern blot results showed 4 of 23 mice were integrated. Two of the founders were positive by RTPCR. Conclusion TNNI2 mutation transgenic mice were successfully established by pronuclear microinjection.