分子技术对表达GFP转基因胚胎的再检测  

Re-detection of Transgenic Embryos by Molecular Technology

作　　者：肖红卫[1] 郑新民[1] 李莉[1] 陈思怀[2] 张涌[2] 魏庆信[1] 

机构地区：[1]湖北省农业科学院畜牧兽医研究所,武汉430064 [2]西北农林科技大学生物工程研究所

出　　处：《湖北农业科学》2006年第2期144-146,共3页Hubei Agricultural Sciences

基　　金：国家863高技术项目资助(2001AA216071)

摘　　要：把经脂质体包埋的含CD59基因的重组质粒pEGFP-CD59注进小鼠睾丸组织和小鼠的输精管中,分别过1个月和7d与经过超排处理的母鼠交配,在胚胎发育的不同发育时期冲出胚胎于荧光显微镜下观察,显出荧光的转基因小鼠胚胎经PCR和Southern杂交检测确认,转基因小鼠胚胎阳性率为11%(3/27)。表明GFP基因不能完全准确地反映转基因的情况,需要用PCR结合Southern杂交方法对表达绿色荧光蛋白转基因胚胎进行再检测。The recombinant plasmids pEGFP-CD59 fused with human CDs9 gene were directly injected into male mouse＇s testis and male mouse＇s spermaductus after combinanted with liposome. The transfected males were fertilized with females one month and seven days later. The embryos at the different stage of development were took out and detected with the Fluorescence Microscope. The positive embryos were confirmed with the PCR and Southern Blotting, the positive rate of transgenic embryos is 11%（3/27）. The GFP gene can＇t confirm transgene completely. The re-detection of transgenic embryos by PCR,Southern Blotting is needed.

关 键 词：PCR SOUTHERN杂交 绿色荧光蛋白 转基因胚胎 

分 类 号：Q781[生物学—分子生物学] Q785