基础医学与临床 ›› 2023, Vol. 43 ›› Issue (3): 415-418.doi: 10.16352/j.issn.1001-6325.2023.03.415

• 研究论文 • 上一篇    下一篇

Nrf2敲除小鼠模型的构建与基因型鉴定

王闽蓉, 张峰, 鲁利群*   

  1. 成都医学院第一附属医院 儿科, 四川 成都 610500
  • 收稿日期:2022-04-15 修回日期:2022-07-21 出版日期:2023-03-05 发布日期:2023-02-27
  • 通讯作者: * luliqun7@sina.com
  • 基金资助:
    成都市科技局技术创新研发项目(2019-YF05-00271-SN)

Construction and genotype identification of Nrf2 knockout mouse model

WANG Minrong, ZHANG Feng, LU Liqun*   

  1. Department of Pediatrics, the First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China
  • Received:2022-04-15 Revised:2022-07-21 Online:2023-03-05 Published:2023-02-27
  • Contact: * luliqun7@sina.com

摘要: 目的 构建并鉴定Nrf2敲除模型小鼠。方法 将引进一对雌雄杂合子(Nrf2-/+)小鼠进行交配,繁育出F1代小鼠;将F1代小鼠中基因型为Nrf2-/+的雌雄小鼠继续进行交配,获得F2代小鼠;小鼠5日龄时剪尾部组织提取DNA,用PCR和琼脂糖凝胶电泳进行基因型结果判定。提取Nrf2敲除小鼠肺组织蛋白质,用Western blot检测NRF2蛋白表达情况,佐证鉴定结果。结果 雌雄纯合子(Nrf2-/-)基因敲除小鼠可继续交配,获得基因敲除纯合子小鼠;F1与F2代杂合子小鼠交配可获得3种基因型:野生型(Nrf2+/+)、杂合子(Nrf2+/-)、纯合子 (Nrf2-/-),用PCR成功鉴定出子代小鼠的基因型;Nrf2敲除小鼠肺组织的NRF2蛋白表达显著低于野生型(P<0.05)。结论 本研究成功繁育出Nrf2全身敲除模型小鼠,为进一步探讨NRF2的作用及其机制提供了新的实验工具。

关键词: Nrf2, 基因鉴定, 肺脏, 小鼠

Abstract: Objective To construct and identify Nrf2 knockout mouse model. Methods A pair of male and female heterozygous (Nrf2-/+) mice were introduced to breed F1 mice. Male and female Nrf2-/+ mice from F1 generation were bred to obtain F2 generation mice. DNA was extracted from the tail tissue of mice at 5 days of age and their genotype was determined by PCR and agarose gel electrophoresis. NRF2 protein was extracted and coding gene was knocked out from the lung tissues of mice. The expression of NRF2 protein was detected by Western blot, and the identification results were confirmed laterally. Results Homozygous (Nrf2-/-) mice continued to mate and obtain homozygous mice. Three genotypes were identified when F1 was mated with F2 generation heterozygous mice: wild type (Nrf2+/+), heterozygous (Nrf2+/-) and homozygous (Nrf2-/-). The genotypes of offspring mice were successfully identified by PCR. NRF2 protein expression in lung tissue of Nrf2 knockout mice was significantly lower than that of wild-type mice (P<0.05). Conclusions The mouse model with Nrf2 knockout is successfully bred in this study, which provides a new experimental tool for further exploring the role and mechanism of NRF2.

Key words: Nrf2, gene identification, lungs, mouse

中图分类号: