精子载体法获得植酸酶转基因猪  

Production of transgenic pigs with phytase gene by sperm-mediated gene transfer

作　　者：吴斌[1,2,3] 戴建军[1,2] 张廷宇[1,2] 张树山[1,2] 吴彩凤[1,2] 顾晓龙[1,2,3] 陈会兰[1,2] 张德福[1,2] 

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106 [2]上海市农业遗传育种重点实验室动物遗传工程研究室,上海201106 [3]四川农业大学生命科学与理学院,雅安625014

出　　处：《上海农业学报》2013年第5期6-9,共4页Acta Agriculturae Shanghai

基　　金：上海市科委业科技成果转化项目(123919N0700;133919N1700);上海市农科院青年科技发展基金(2011-07);国家转基因生物新品种培育科技重大专项(2008ZX08006-005;2009ZX08006-014B)资助

摘　　要：将带有植酸酶基因的重构质粒酶切纯化后,与去除精浆的猪新鲜精液按比例混合,17℃孵育处理使重构质粒结合到精子头部,通过人工授精对6头受体母猪进行配种,4头受孕,产仔38头。经PcR对特异性片段扩增,结果表明:有6头PcR结果为阳性,阳性率为15.8%。扩增片段经克隆、测序、比对,与阳性对照组序列相似率均在99%以上,说明目的基因已转入阳性个体体内。After digested and purified,the recombinant plasmid with phytase genes was mixed pro- portionally with the fresh seminal fluid whose seminal plasma was removed, and then incubated at 17 ℃ to make the plasmid combine with the spermhead. The joining with 6 sows was carried out by ar- tificial insemination, producing 4 pregnant sows and farrowing 38 piglets. PCR amplification of the specific fragments showed that the PCR products of 6 piglets were positive, and the cloning, sequen- cing and alignment indicated that all the rates of sequence similarity between the amplified fragments and the control group were over 99% ,demonstrating that the phytase gene had been transferred into the positive piglets.

关 键 词：精子载体法 植酸酶基因 转基因猪 

分 类 号：S828[农业科学—畜牧学] S813.8[农业科学—畜牧兽医]