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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

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【作者】 肖红卫郑新民李莉陈思怀张涌魏庆信

【Author】 XIAO Hong-wei1,2,ZHENG Xin-min1,LI Li1,CHEN Si-huai2,3,ZHANG Yong2,WEI Qing-xin1(1.Institute of Animal and Veterinary Science,Hubei Academy of Agricultural Sciences,Wuhan 430064,China;2.Institute of Bioengineering,Northwest Sci-Tech University of A& F,Yangling 712100,Shannxi,China;3.College of Anamal Science and Technology,Southwest Agricultural University,Rongchang 400716,Chongqing,China)

【机构】 湖北省农业科学院畜牧兽医研究所西北农林科技大学生物工程研究所湖北省农业科学院畜牧兽医研究所 武汉430064陕西杨凌712100武汉430064西南农业大学动物科技学院重庆荣昌400716

【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。

【Abstract】 The recombinant plasmids pEGFP-CD59 fused with human CD59 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】 PCRSouthern杂交绿色荧光蛋白转基因胚胎
【Key words】 PCRSouthern blottingGFPtransgenic embryos
【基金】 国家863高技术项目资助(2001AA216071)
  • 【文献出处】 湖北农业科学 ,Hubei Agricultural Sciences , 编辑部邮箱 ,2006年02期
  • 【分类号】Q813.7;Q78
  • 【下载频次】113
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