实验动物科学 ›› 2024, Vol. 41 ›› Issue (6): 82-86.DOI: 10. 3969 / j. issn. 1006-6179. 2024. 06. 014

• 论著 • 上一篇    下一篇

C3 基因过表达模型小鼠的鉴定及饲养繁育

  

  1. ( 1. 首都医科大学实验动物部,北京 100069)  ( 2. 首都医科大学基础医学院实验动物学学系,北京 100069)
    ( 3. 首都医科大学基础医学院神经再生修复研究北京市重点实验室,北京 100069)
  • 收稿日期:2024-02-23 出版日期:2024-12-28 发布日期:2025-01-15
  • 通讯作者: 卢 静( 1969—) ,教授,博士研究生导师,研究方向为生殖发育,E-mail: lujing@ ccmu. edu. cn。 仵 毅( 1982—) ,副教授,研究方向为生殖发育,E-mail: wuyi@ ccmu. edu. cn。
  • 作者简介:彭博雅( 1984—) ,主管技师,研究方向为生殖发育,E-mail: pengboya@ ccmu. edu. cn。

Construction and Breeding of C3 Gene Over-expression Model Mice

  1. ( 1. Department of Experimental Animals,Capital Medical University, Beijing 100069,China)
    ( 2. Department of Experimental Zoology,School of Basic Medicine,Capital Medical University,Beijing 100069,China)
    ( 3. Beijing Key Laboratory of Nerve Regeneration and Repair,School of Basic Medicine,Capital Medical University,Beijing 100069,China)
  • Received:2024-02-23 Online:2024-12-28 Published:2025-01-15

摘要:

目的 构建并繁育 C57BL / 6J-ROSA26 em( C3+) 基因过表达小鼠,鉴定并分析其基因型和目的蛋白的表达情况,并验证检测方法的可适用性。 方法 将构建的 C57BL / 6J-ROSA26 em( C3+) 基因过表达阳性 founder 鼠与野生型小鼠配繁,获得的杂合子小鼠采取同胞交配方式(雌雄比 1 ∶ 1) ,得到纯合子小鼠。 使用鼠尾组织 DNA PCR 法鉴定杂合子及纯合子基因型,通过 Western blot 法检测小鼠间脑组织 C3 蛋白的表达差异。 分析 C57BL / 6J-ROSA26 em( C3+)基因过表达小鼠杂合子及纯合子的体质量变化和繁殖性能。 结果 C57BL / 6J-ROSA26 em( C3+) 基因过表达杂合子小鼠通过同胞交配可获得纯合子小鼠。 蛋白印迹法检测发现,与野生型小鼠相比,C3 蛋白在基因修饰小鼠组织内高表达。 C3 基因的过表达不影响小鼠繁殖,纯合子间可以稳定配繁。 但是,C3 基因的过表达显著降低了 5 周龄后转基因雌鼠的体 质 量。结论采用全同胞交配方式, 并通过PCR法进行基因型鉴定, 可稳定获得C57BL / 6J-ROSA26 em( C3+) 基因过表达小鼠。

关键词:

Abstract: Objective C3 gene over-expression mice were constructed and bred to identify and analyse their genotypes and the expression of target proteins, and to verify the applicability of the assay. Method The constructed C3 gene over-expression positive founder mice were bred with wild-type mice, and the heterozygous mice were obtained by sibling mating ( male-to-female ratio of 1 ∶ 1 ) , and the pure heterozygous mice were obtained. The genotypes of heterozygotes and heterozygotes were identified by PCR of mouse tail tissue DNA, and the differences in C3 protein expression in brain tissue between mice were detected by Western blot. The body weight changes and reproductive performance of heterozygous and pure heterozygous C3 over-expressing mice were analyzed. Result C3 gene over-expression heterozygous mice could obtain pure heterozygous mice by sibling mating. Protein blotting showed that C3 protein was highly expressed in the tissues of transgenic mice compared with wild-type mice, and the over-expression of C3 gene did not affect the reproduction of mice, and the mating was stable among the homozygous mice. However, over-expression of the C3 gene significantly reduced the body weight of transgenic female mice after 5 weeks of age. Conclusion The C3 over-expression mice can be stably obtained by full-sib mating and genotyping by PCR.

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