突变靶基因xylE转基因小鼠

Transgenic Mice Carrying xylE Mutational Target Gene

本研究以ｐＥＳｎｘ穿梭质粒为载体，选用恶臭假单孢杆菌ＴＯＬ质粒上的ｘｙｌＥ基因作为突变靶基因组构建的重组构件，经微注射导入ＩＣＲ小鼠５４９枚受精卵雄原核，将存活的３５２枚卵移入２４只假孕鼠输卵管内，７只妊娠，共产仔４１只，其中７只死胎，出生后死亡４只，３０只存活。注射卵存活率为６４％（３５２／５４９），出生率为１１．６％（４１／３５２）。存活鼠经ＰＣＲ－Ｓｏｕｔｈｅｒｎ检测，整合率为５７％（１７／３０）。在整合阳性鼠中选择了杂交信号较强的２只当代公鼠，通过回收载体和转化试验，结果表明整合基因完整，具有转化的功能，由此，建立了突变靶基因ｘｙｌＥ转基因小鼠模型。

In this study,transgenic mice were generated by microinjecting pESnx shuttle vector (12.7Kb) into the male pronucleus of 549 fertilized eggs of ICR mice.The pESnx contains a xylE mutational target gene from TOL plasmid of pseudomona putida.352 eggs survived were transplanted into the oviduct of 24 psuedopregnant ICR mice respectively. 7 got pregnant and 41 offsprings were delivered,7 of them were stillbirths,4 of them died in few days after birth.The ratio of survival eggs is 64%(352/549) and the ratio of birth is 11.7 offsprings per 100 transfered eggs.17 positive of the 30 survival offsprings were identified by PCR and Southern blotting analysis,the ratio of integration is 57%(17/30).By recovery of the vector and transformation,it indicated the integrated vector contained intact target gene,so transgenic mouse model carrying xylE mutational target gene was established.