新生血管可视化转基因小鼠的建立

摘要

目的 建立新生血管的内皮中有绿色荧光蛋白(GFP)表达的转基因动物.方法 构建含巢蛋白第二内含子和GFP的载体nestin-hsp68-gfp,转入U251细胞,验证其在真核细胞中的表达.然后用显微注射的方法,建立转基因动物.小鼠出生后,剪尾提基因组,用PCR初筛,然后对初筛阳性的小鼠检测GFP的表达.结果 出生13只小鼠,初筛2只阳性.能检测到GFP在出生后和胚胎小鼠中有表达.结论 成功制作标记新生血管的转基因小鼠.%Objective To establish transgenic mice with GFP expression in the vascular endothelium during neovascularization. Methods The vector nestin-hsp68-gfp containing nestin second inrron was introduced into U251 cells and the expression level of GFP was detected by fluorescence microscopy. Transgenic mice were produced by microinjection. The genome of the offspring mice was screened by PCR, and GFP expression in the vascular endothelium was detected using immunohistochemistry. Results Thirteen offspring mice were obtained and 2 of them were positive for GFP in the vascular endothelium as detected by PCR. GFP was detected in the offspring mice both at the embryonic stage and after birth. Conclusion The transgenic mice with GFP expression in the vascular endothelium during neovascularization have been successfully established.