TNNT3^(R69H)突变转基因小鼠模型的建立  

Establishment of TNNT3^(R69H) Mutation Transgenic Mice

作　　者：李兆阳[1] 吕相川[1] 汪瑛[1] 杨葳[1] 于洋[1] 周生来[1] 王惟[1] 张梅英[1] 郑志红[1] 

机构地区：[1]中国医科大学实验动物部辽宁省转基因动物重点实验室,辽宁沈阳110001

出　　处：《中国比较医学杂志》2012年第2期9-13,共5页Chinese Journal of Comparative Medicine

基　　金："重大新药创制"科技重大专项2010ZX09401-304;辽宁省科技厅攻关项目2009408001-1

摘　　要：目的建立TNNT3(R69H)突变转基因小鼠模型。方法构建pEGFP-TNNT3(R69H)转基因构件,通过原核显微注射方法将线性化、纯化后的外源质粒pEGFP-TNNT3(R69H)注射入BDF1小鼠受精卵中,胚胎移植至同期发情的假孕受体母鼠输卵管内,获得子代小鼠。用PCR和Southern blot方法检测子代鼠尾基因组DNA,通过RT-PCR及Western blot的方法检测TNNT3基因表达。结果 8只假孕小鼠共移植注射后的胚胎82枚,出生40只子代鼠,经PCR和Southern方法检测得到5只转基因阳性小鼠。对其子代小鼠进行RT-PCR、Western blot检测结果显示,TNNT3在转基因小鼠心脏和骨骼肌中表达量明显增多。结论通过显微注射法使外源基因pEGFP-TNNT3(R69H)在小鼠基因组中整合,成功建立了TNNT3(R69H)突变转基因小鼠模型。Objective To establish an animal model of TNNT3（（R69H）） mutation transgenic mice.Methods Transgenic component pEGFP-TNNT3（R69H） was constructed,linearized and purified,then microinjected into fertilized mouse eggs.These eggs were transplanted into pseudopregant mice.The genotype of transgenic founders were identified by PCR and Southern blot.The expression of TNNT3（R69H） transcript in the tissues of the transgenic mice was detected by RT-PCR and Western blot.Results Transgenic BDF1 mice were obtained by microinjection.PCR and Southern blot results showed 5 of 40 mice were integrated.Conclusions TNNT3（（R69H）） mutation transgenic mouse models are successfully established by pronuclear microinjection.

关 键 词：转基因小鼠 TNNT3突变 显微注射 

分 类 号：R33[医药卫生—人体生理学]