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.
出处
《上海农业学报》
CSCD
北大核心
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
关键词
精子载体法
植酸酶基因
转基因猪
Sperm-mediated gene transfer
Phytase gene
Transgenic pig
