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背景与目的 Vegfr2-luc转基因小鼠体内血管内皮生长因子受体2(vascular endothelial growth factor recep- tor 2,VEGFR2)的表达可以驱动荧光素酶报告基因(luciferase,luc)的表达,是活体动物水平实时监测血管生成情况的有利工具。本研究旨在对子代Vegfr2-luc转基因小鼠进行鉴定,以确定能否用于血管生成研究。方法 PCR检测新生小鼠基因组内luc基因;利用活体成像技术观察新生Vegfr2-luc转基因小鼠生长发育过程中以及皮肤伤口修复过程中 luc基因表达水平的变化情况;荧光素酶报告基因检测试剂盒检测成年(8周龄)转基因小鼠各脏器荧光素酶的活性和Real-time PCR检测各器官VEGFR2 mRNA的表达水平。结果 PCR结果显示50%(56/112)的新生小鼠携带luc基因。活体成像结果显示随着Vegfr2-luc转基因小鼠发育成熟,luc表达量逐渐降低(P<0.001);在皮肤伤口修复过程中, 伤口处luc表达水平先增强后降低(P<0.001)。雌性成年转基因小鼠各脏器VEGFR2 mRNA的表达水平与荧光素酶活性呈正相关(r=0.948,P<0.001)。将睾丸组织除外,雄性成年转基因小鼠各脏器VEGFR2mRNA的表达水平与荧光素酶活性同样呈正相关(r=0.836,P<0.001)。结论 Vegfr2-luc转基因子代小鼠体内luc表达水平的变化可以反映VEGFR2 的表达情况。
BACKGROUND & OBJECTIVE: The expression of luciferase (luc) gene in vegfr2-luc transgenic mice is driven by the expression of vascular endothelial growth factor receptor 2 (VEGFR2) Good tool to monitor angiogenesis in real time. The aim of this study was to identify progeny Vegfr2-luc transgenic mice to determine whether they can be used in angiogenesis studies. Methods The luc gene was detected by PCR in vivo. The changes of luc gene expression in the neonatal Vegfr2-luc transgenic mice during the growth and wound repair were observed by live imaging. The luciferase reporter assay kit The luciferase activity of each organ in adult (8 weeks old) transgenic mice and the expression of VEGFR2 mRNA in each organ were detected by Real-time PCR. Results PCR results showed that 50% (56/112) of newborn mice carried the luc gene. The results of in vivo imaging showed that the expression of luc decreased gradually with the development of Vegfr2-luc transgenic mice (P <0.001). At the wound healing, the expression of luc in the wounds increased first and then decreased (P <0.001). The expression of VEGFR2 mRNA in the organs of female adult transgenic mice was positively correlated with the luciferase activity (r = 0.948, P <0.001). Excluding the testicular tissue, the expression of VEGFR2 mRNA in the organs of male adult transgenic mice was also positively correlated with the luciferase activity (r = 0.836, P <0.001). Conclusion The changes of luc expression level in Vegfr2-luc transgenic offspring can reflect the expression of VEGFR2.