›› 2009, Vol. 36 ›› Issue (7): 72-74.

• 生物技术 • 上一篇    下一篇

缺失糖基磷脂酰肌醇锚的Doppel 转基因小鼠的构建

李静1,张洋2,于恒智3,赵魁1,张守峰2,扈荣良2,邓旭明1   

  1. (1.吉林大学畜牧兽医学院, 长春 130062; 2.军事医学科学院军事兽医研究所, 长春 130062;3.青岛农业大学, 青岛 266109)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-07-20 发布日期:2009-07-20
  • 通讯作者: 邓旭明

Foundation of Transgenic Mice Expressing Doppel without Glycosyl Phosphatidyl Inositol

LI Jing1, ZHANG Yang2, YU Heng-zhi3, ZHAO Kui1, ZHANG Shou-feng2,HU Rong-liang2, DENG Xu-ming1   

  1. (1.Department of Animal Husbandry and Veterinary Medicine, Jilin University, Changchun 130062, China; 2.Veterinary Research Institute, Academy of Military Medical Science, Changchun 130062, China;3.Qingdao Agricultural University, Qingdao 266109, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-07-20 Published:2009-07-20
  • Contact: DENG Xu-ming

摘要: 将缺失糖基磷脂酰肌醇(glycosyl phosphatidyl inositol,GPI)的Doppel的质粒(pDoppel 1-155),用Not Ⅰ酶切,回收并纯化目的基因片段,通过显微注射,导入BALB/c小鼠受精卵的雄原核内,结果32只仔鼠出生,其中有6只PCR检测阳性。将其中1只F0代小鼠与正常小鼠交配,共获得21只F1代小鼠,经PCR检测,有10只仔鼠为阳性。对2只F1代PCR阳性小鼠脑组织进行RT-PCR检测,其中1只为阳性。采集3只F1代PCR阳性小鼠脑组织进行Western blotting检测,在小鼠脑组织能够表达17 ku的Doppel 1-155蛋白。本研究为进一步深入研究Doppel蛋白在体内的生物学特性提供了转基因动物模型。

关键词: 糖基磷脂酰肌醇; Doppel; 转基因; 小鼠

Abstract: A recombinant Doppel plasmid without glycosyl phosphatidyl inositol (pDoppel 1-155) was digested by Not Ⅰ,then the objective gene was recovered and purified. The gene was introduced into male pronucleus of BALB/c mouse fertilized ovum by microinjection, then we got 32 mice which have been injected the Dpl 1-155 gene, in which 6 was positive by PCR; one F0 generation of Tg mouse expressing Dpl without GPI was mated with a wild-type mouse, then 21 F1 generation mouse were got, in which 10 were positive by PCR identification. Brain tissue of three F1 generation mice which were positive by PCR identification was collected, and was identified by Western blotting, there was 17 ku Dpl 1-155 protein expressed in the brain. So we have generated Tg mice expressing Dpl without GPI which provides Tg animal models for investigating the bionomics of Dpl protein.

Key words: glycosyl phosphatidyl inositol; Doppel; transgene; mouse

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