中国医科大学学报

中国医科大学学报

中国医科大学学报 ›› 2009, Vol. 38 ›› Issue (11): 820–.

• 基础医学 • 上一篇    下一篇

突变DJ-1转基因小鼠模型的建立

张梅英; 吴红联; 蔺美娜; 李兆阳; 周生来; 于洋; 王惟; 吕相川; 王禄增   

  1. 中国医科大学实验动物中心
  • 出版日期:2009-12-20 发布日期:2010-04-14
  • 通讯作者: 王禄增 E-mail:wanglz@163.com

Construction of Transgenic Mice Carrying DJ-1 Mutation

  • Online:2009-12-20 Published:2010-04-14

摘要:

目的探讨DJ-1L166P突变基因在帕金森病发生中的作用。方法构建pcDNA3.1-myc-his-DJ-1L166P真核表达载体,利用显微注射方法将DJ-1L166P基因片段(约3.9kb)导入BDF1小鼠受精卵雄原核并移植到同期受孕的ICR假孕母鼠输卵管中,对产出仔鼠的鼠尾组织DNA进行聚合酶链反应和Southern
      blot检测,对Southern blot鉴定阳性的转基因小鼠进行逆转录聚合酶链反应和Western
      blot检测。结果共产生20只子代小鼠,1号和7号为DJ-1L166P转基因阳性小鼠。1号转基因小鼠大脑、小脑、肝、肺、肾、睾丸有不同程度的DJ-1mRNA表达,均高于正常对照小鼠,但只有大脑中DJ-1mRNA的表达水平显著高于正常对照小鼠(P<0.05);7号转基因小鼠只有肝、肺组织中有DJ-1mRNA表达,也高于正常对照小鼠,但差异无统计学意义。只有1号小鼠大脑有His标签蛋白的表达。结论成功获得1只DJ-1L166P基因突变转基因小鼠模型,为课题的下一步实验研究打下良好的基础。

关键词: DJ-1基因, 帕金森症, 转基因小鼠, DJ-1, 帕金森症, 转基因小鼠

Abstract:

Objective To study the role of DJ-1L166P mutation in Parkinson
      Disease.Methods pcDNA3.1-myc-his-DJ-1 L166P was constructed and then
      microinjected into mouse fertilized eggs.These eggs were transplanted into
      the oviduct of pseudopregnant mice.Transgenic mice were identified by
      polymerase chain reaction and Southern blot.Reverse transcription
      plolymerase chain reaction and Western blot were used to detect gene
      transcription and expression.Results A total of 20 transgenic mice were
      obtained,and 2 of these 20 mice(No.1 and No.7 mice,respectively)were
      integrated succesfully.In No.1 mouse,the expression of DJ-1 mRNA was
      detected in the cerebrum,cerebellum,liver,lung,kidney,and testicle.The
      level of DJ-1 mRNA in the cerebrum was higher in No.1 mouse than in
      controls(P < 0.05).The expression of DJ-1 mRNA was detected in the liver
      and lung of No.7 mouse,but no significant difference was found in the
      level of DJ-1 mRNA between No.7 mouse and controls.Histone tag was
      expressed only in the cere...更多brum of No.1 mouse.Conclusion The DJ-1L166P
      transgenic mouse model was successfully established,and it provides a good
      basis for further research.

Key words: DJ-1, Parkinson disease, transgenic mouse, DJ-1, Parkinson’s Disease, transgenic mice

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