血管内皮细胞敲除Rheb基因杂合子小鼠模型建立及意义

发布时间：2018-06-28 10:49

  本文选题：基因敲除 + Cre/LoxP条件性基因敲除技术　； 参考：《山东医药》2017年36期

【摘要】：目的建立血管内皮细胞敲除Rheb基因杂合子小鼠模型,旨在进一步研究Rheb基因在胚胎血管形成中的作用。方法将5只Rheb~(flox/flox)雄鼠与15只Rheb~(flox/+)雌鼠进行交配,共繁殖出子代小鼠76只(基因型为Rheb~(flox/flox)或Rheb~(flox/+))。将76只子代小鼠与20只血管内皮细胞特异性表达Tek-Cre重组酶的小鼠(以下称Cre小鼠)进行交配,得到基因型为Tek-Cre×Rheb~(flox/+)及单纯Rheb~(flox/+)子代小鼠共140只。采用PCR法对140只小鼠鼠尾组织Rhebflox及Tek-Cre基因型进行鉴定,并记录其2月龄体长及体质量;采用Western blotting法检测Tek-Cre×Rheb~(flox/+)小鼠与单纯Rheb~(flox/+)小鼠肝组织Rheb蛋白表达。结果共得到基因型为Tek-Cre×Rheb~(flox/+)小鼠72只、单纯Rheb~(flox/+)小鼠68只,其体长分别为(18.50±0.13)、(18.80±0.10)cm,体质量分别为(29.50±0.15)、(30.10±0.21)g,二者比较P均0.05;肝组织Rheb蛋白相对表达量分别为0.21±0.17、0.66±0.12,二者比较P0.05。结论成功构建血管内皮细胞敲除Rheb基因杂合子小鼠模型,将为研究Rheb基因在胚胎血管形成中的作用奠定基础。
[Abstract]:Objective to establish a murine model of vascular endothelial cell knockout Rheb gene heterozygote in order to further study the role of Rheb gene in embryonic angiogenesis. Methods five male flox/flox rats and 15 female Rheb ~ (flox/) mice were mated, and 76 offspring mice (genotype Rheb~ (flox/flox) or flox/).) were bred. 76 offspring mice were mated with 20 mice with Tek-Cre recombinant enzyme specifically expressed by vascular endothelial cells (hereinafter referred to as CRE mice). The genotypes of Tek-CRE 脳 Rheb~ (flox/) and pure Rheb~ (flox/) progenies were 140. The Rhebflox and Tek-Cre genotypes in the tail tissues of 140 mice were identified by polymerase chain reaction (PCR), and their body length and body weight were recorded, and the expression of Rheb protein in the liver tissues of Tek-Cre 脳 Rheb~ (flox/) mice and simple Rheb~ (flox/) mice were detected by Western blotting. Results there were 72 Tek-Cre 脳 Rheb~ (flox/) mice and 68 simple Rheb~ (flox/) mice with body length (18.50 卤0.13), (18.80 卤0.10) cm and body weight (29.50 卤0.15), (30.10 卤0.21) g, respectively (P 0.05), and the relative expression of Rheb protein in liver tissue was 0.21 卤0.170.66 卤0.12, respectively (P 0.05). Conclusion the successful construction of vascular endothelial cell knockout mouse model of Rheb gene heterozygote will lay a foundation for the study of the role of Rheb gene in embryonic angiogenesis.
【作者单位】： 南方医科大学南方医院;湖南医药学院第一附属医院;南方医科大学武汉临床医学院;
【基金】：广州市科技计划项目(2013J4500040)
【分类号】：R321;R-332
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