RFP转基因小鼠模型的建立

发布时间：2018-07-04 07:33

本文选题：红色荧光蛋白(RFP) + 载体　；参考：《黑龙江畜牧兽医》2017年19期

【摘要】：为了建立组织广泛表达红色荧光蛋白(red fluorescence protein,RFP)的转基因小鼠,为肿瘤和干细胞等研究提供动物模型,试验从pdsred-express2-n1载体上酶切后获得线性化的CMV-Dsred基因片段,将CMV-Dsred基因注入ICR小鼠受精卵雄原核内,获得的仔鼠利用活体荧光蛋白观察系统、PCR和Southern-blot的方法进行分子水平的检测,同时用后代小鼠的组织制作冰冻切片在荧光显微镜下观察。结果表明:荧光蛋白活体成像系统检测到组织广泛表达红色荧光蛋白的小鼠,阳性小鼠基因组中均检测到Dsred基因的整合,在心脏、肝脏、脾脏、肺脏、肾脏、肠道、子宫、肌肉、脂肪和皮肤等组织器官中均检测到红色荧光蛋白。说明成功获得了组织广泛表达红色荧光蛋白转基因小鼠模型。
[Abstract]:In order to establish transgenic mice with widely expressed red fluorescent protein (red fluorescence protein (RFP), and to provide animal models for tumor and stem cell studies, a linearized CMV-Dsred gene fragment was obtained from pdsred-express2-n1 vector after restriction endonuclease digestion. CMV-Dsred gene was injected into the male pronucleus of ICR mouse fertilized eggs. The offspring were detected at molecular level by PCR and Southern-blot, and frozen sections were made from the tissues of progeny mice under fluorescence microscope. The results showed that in vivo fluorescence imaging system, the tissues of mice with widely expressed red fluorescent protein were detected, and the integration of Dsred gene was detected in the genomes of positive mice, including heart, liver, spleen, lung, kidney, intestine, uterus, liver, spleen, lung, kidney, intestine, uterus, kidney, intestine and uterus. Red fluorescent protein was detected in muscle, fat and skin. The results showed that the transgenic mouse model of widely expressing red fluorescent protein was successfully obtained.
【作者单位】：华南农业大学动物科学学院广东省农业动物基因组学与分子育种重点实验室;河南牧业经济学院动物科技学院;
【基金】：国家“973”计划专项(2011CB944202) 河南牧业经济学院博士科研启动基金项目(51000788)
【分类号】：Q78;S865.13

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