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用显微注射法建立转bcl-x_l基因小鼠

Developing bcl-x_l transgenic mice by microinjection

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【作者】 肖文伍王芙蓉张苏明赵浩斌郑新明樊俊华魏庆信张旻

【Author】 XIAO Wen wu 1 , WANG Fu rong 1 ,ZHANG Su ming 1 ,ZHAO Hao bin 2 , ZHENG Xin ming 2 , FAN Jun hua 2 , WEI Qing xin 2 , ZHANG Ming 1 (1.Department of neurology, Tongji Hospital,Tongji Medical College,Huazhong Scien

【机构】 华中科技大学同济医学院附属同济医院神经科湖北省农业科学院生物工程室华中科技大学同济医学院附

【摘要】 建立转bcl xl基因小鼠 ,继而传代建系 ,用于缺血性脑血管疾病的研究。采用显微注射法 ,将人bcl xl基因注入昆明小白鼠受精卵获得子代鼠 ,然后作PCR ,Southern blot ,mRNA ,Western blot检测以获得阳性鼠。实验中注射受精卵 16 5 4枚 ,移植卵数 14 4 8枚 ,受体鼠 4 9只 ,怀孕鼠 7只 ,子代鼠 13只 ,整合数 4只并均有表达 ,植入受精卵的存活率 83% ,受精卵的总存活率 0 .78% ,移植鼠的怀孕率 14 .3% ,整合效率 30 .7% ,总有效率为 0 .2 4 %。初步获得了转bcl xl基因小鼠

【Abstract】 It was attempted to develop bcl x l transgenic mouse and produce its lineages for studying ischemic cerebrovascular diseases,bcl x l gene was introduced into mice by microinjection .The presence of bcl x l and its expression were confirmed by PCR,Southern blot,mRNA inspection,Western blot in the founders.In our experiments, bcl x l gene was injected into pronucleus of 1 654 zygotes and 1 448 injected eggs were implanted into 49 foster mothers, 7 of which were pregnant and bore 13 offsprings. Among the babies, 4 offsprings were integrated with the bcl x l gene and expressed it. Totally, implanted zygotes survival rate (83%), zygotes total survival rate (0.78%), recipient mouse pregnance rate ( 14.3% ), integration rate (30.7%), expression rate(100%) ,total efficiency(0.24%)were obtained respectively. Bcl x l transgenic mice were obtained, 4 in number, but the transgenic efficiency was very low and the expression was unstable. The model of cerebral vascular disease will be made after acquiring transgenic lineages.

【关键词】 转基因小鼠bcl-xl基因显微注射
【Key words】 transgenic mousebcl x l genemicroinjection
【基金】 国家自然科学基金重点项目 (3 973 0 170 ) ;国家自然科学基金面上项目 (3 9770 810 )
  • 【文献出处】 中国神经科学杂志 ,Chinese Journal of Neuroscience , 编辑部邮箱 ,2002年03期
  • 【分类号】R743.31
  • 【被引频次】12
  • 【下载频次】178
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