牙本质涎蛋白转基因小鼠的构建

Generation of transgenic mouse of dentin sialoprotein

目的构建小鼠牙本质涎蛋白(DSP)转基因小鼠。方法将pcDNA3.1载体中的CMV启动子替换为启动子cβ-actin,构建载体pcDNA3.1-CX,然后将PCR获得的DSP基因编码序列克隆到pcDNA3.1-CX中,构建DSP转基因载体pcDNA3.1-CX-dsp;将线性化的载体DNA注射到小鼠受精卵的雄原核,受精卵移植到假孕母鼠的输卵管。仔鼠出生后,用PCR及Southern blot检测阳性小鼠。结果共移植717枚注射过的受精卵至29只受体鼠,移卵后产仔67只,阳性4只。阳性鼠分别传代,开始建系。结论通过显微注射的方法成功获得了DSP转基因小鼠。

Objective To establish the transgcnic mouse model of DSP. Methods Plasrnid pcDNA3.1 - CX was constructed by substituting promoter cβ - actin for CMV promoter of pcDNA3.1. And the ultimate tranagenic vector, peDNA3.1 - CX - dsp, was constructed by cloning DSP coding sequence into peDNA3.1 - CX. The peDNA3.1 - CX - dsp plasmid was linearized and mieroinjeeted into the male pronueleus of the zygotes. The tail DNA of pups was tested by PCR and Southern blot.Results Seven hundred and seventeen （717） embryos were implanted to twenty-nine recipient pseudopregnant mice. Four of the sixty - seven pups carried the transgene. The establishment of the transgenic mouse line of DSP was under progress. Conclusion Founders of the DSP transgenic mouse were obtained successfully.