本文選題：基因敲除 + Cre/LoxP條件性基因敲除技術(shù)　； 參考：《山東醫(yī)藥》2017年36期

【摘要】：目的建立血管內(nèi)皮細(xì)胞敲除Rheb基因雜合子小鼠模型,旨在進(jìn)一步研究Rheb基因在胚胎血管形成中的作用。方法將5只Rheb~(flox/flox)雄鼠與15只Rheb~(flox/+)雌鼠進(jìn)行交配,共繁殖出子代小鼠76只(基因型為Rheb~(flox/flox)或Rheb~(flox/+))。將76只子代小鼠與20只血管內(nèi)皮細(xì)胞特異性表達(dá)Tek-Cre重組酶的小鼠(以下稱Cre小鼠)進(jìn)行交配,得到基因型為Tek-Cre×Rheb~(flox/+)及單純Rheb~(flox/+)子代小鼠共140只。采用PCR法對140只小鼠鼠尾組織Rhebflox及Tek-Cre基因型進(jìn)行鑒定,并記錄其2月齡體長及體質(zhì)量;采用Western blotting法檢測Tek-Cre×Rheb~(flox/+)小鼠與單純Rheb~(flox/+)小鼠肝組織Rheb蛋白表達(dá)。結(jié)果共得到基因型為Tek-Cre×Rheb~(flox/+)小鼠72只、單純Rheb~(flox/+)小鼠68只,其體長分別為(18.50±0.13)、(18.80±0.10)cm,體質(zhì)量分別為(29.50±0.15)、(30.10±0.21)g,二者比較P均0.05;肝組織Rheb蛋白相對表達(dá)量分別為0.21±0.17、0.66±0.12,二者比較P0.05。結(jié)論成功構(gòu)建血管內(nèi)皮細(xì)胞敲除Rheb基因雜合子小鼠模型,將為研究Rheb基因在胚胎血管形成中的作用奠定基礎(chǔ)。
[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 鹵0.13), (18.80 鹵0.10) cm and body weight (29.50 鹵0.15), (30.10 鹵0.21) g, respectively (P 0.05), and the relative expression of Rheb protein in liver tissue was 0.21 鹵0.170.66 鹵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.
【作者單位】： 南方醫(yī)科大學(xué)南方醫(yī)院;湖南醫(yī)藥學(xué)院第一附屬醫(yī)院;南方醫(yī)科大學(xué)武漢臨床醫(yī)學(xué)院;
【基金】：廣州市科技計劃項目(2013J4500040)
【分類號】：R321;R-332
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本文編號：2077746