β-肌动蛋白启动子指导下人bcl-2转基因小鼠模型的建立

Establishment of transgenic mouse model carrying bcl-2 under the direction of β-actin promoter

目的 :通过原核注射法将肌动蛋白启动子 /bcl 2基因转入小鼠受精卵 ,制作转bcl 2基因的小鼠 .方法 :构建 β actin启动子 /bcl 2表达载体 ,小鼠进行超排获得原核期胚胎 ,应用原核注射法进行转基因操作 ,对新生小鼠通过基因组DNAPCR和Southernblot方法进行鉴定 .结果 :注射成功率为5 8% (5 80 / 10 0 0 ) ,新生小鼠为 33只 ,PCR检测转基因阳性为 8只 ,经过Southern杂交检测有 4只转基因阳性鼠 .结论 :通过原核注射法获得转bcl 2基因的阳性小鼠 ,建立bcl 2转基因动物模型 ,为脑缺血再灌注损伤的机制研究提供手段 .

AIM: To construct a transgenic mouse model carrying bcl-2 gene by prokaryotic injection of promoter-β-actin-bcl-2. METHODS: The promoter-β-actin-bcl-2 expression vector was constructed and the vector was injected into mouse prokaryotic embryo and the DNA of transgenic mice was identified with PCR and Southern blot. RESULTS: The achievement rate of prokaryotic injection was 58%. Among the 33 newborn mice, 8 were tested to be bcl-2 positive with PCR while Southern blotting identified only 4 bcl-2 positive mice. CONCLUSION: The transgenic mouse model carrying bcl-2 gene has been established by prokaryotic injection, which provides an effective model in studying the mechanism of cerebral ischemia-reperfusion injury.