›› 2011, Vol. 38 ›› Issue (11): 73-76.

• 生物技术 • 上一篇    下一篇

转人乳铁蛋白基因奶牛多重PCR检测方法的研究

朱振营1, 李风翠1,2, 林祥梅1, 吴绍强1, 仇松寅1, 李宁3, 王建武3, 薛振华4, 刘建1, 张书霞2   

  1. 1. 中国检验检疫科学研究院动植物检疫研究所,北京 100029;2. 南京农业大学,南京 210095;3. 中国农业大学农业生物技术国家重点实验室,北京 100193;4. 北京市畜牧兽医总站,北京 100107
  • 收稿日期:2011-08-25 修回日期:2011-08-25 出版日期:2011-11-20 发布日期:2011-11-20
  • 通讯作者: 刘建,张书霞

Multiplex PCR Detection of Exogenous Gene in hLF Transgenic Cow

ZHU Zhen-ying1, LI Feng-cui1,2, LIN Xiang-mei1, WU Shao-qiang1, QIU Song-yin1, LI Ning3, WANG Jian-wu3, XUE Zhen-hua4, LIU Jian1, ZHANG Shu-xia2   

  1. 1. Institute of Animal and Plant Quarantine, Chinese Academy of Inspection and Quarantine,Beijing 100029,China;2. Nanjing Agricultural University,Nanjing 210095,China;3. State Key Laboratory for Agrobiotechnology,China Agricultural University,Beijing 100193,China;4. Beijing Municipal Animal Husbandry and Veterinary General Station,Beijing 100107,China
  • Received:2011-08-25 Revised:2011-08-25 Online:2011-11-20 Published:2011-11-20

摘要: 本研究旨在建立转基因奶牛多重PCR快速检测方法,为转基因动物及产品出入境检测技术平台的建立提供技术支持,并为转基因动物及产品检测技术标准的制定提供参考。根据牛物种特异性基因(mtDNA)设计奶牛内源基因引物,根据外源基因人乳铁蛋白基因(human lactoferrin,hLF)和新霉素磷酸转移酶基因(NPT Ⅱ)设计特异性引物,优化反应条件,建立转基因奶牛多重PCR检测方法。本方法敏感、快速、特异,一个反应可以检测多个基因片段,可有效用于转基因奶牛外源基因的检测。

关键词: 转基因奶牛; 多重PCR; hLF基因

Abstract: For developing the typical testing standard of transgenic mammals and its derived food and the technical support for entry﹠exit animal quarantine, detection of the transgenic component in transgenic cow by multiplex PCR was developed in this study. The endogenous mtDNA gene specific for cow identification, exotic hLF gene,marker NPTⅡ gene were amplified according to different primers. The process of the multiplex PCR reaction was optimized. The system described herein represent simple, accurate, and sensitive detection methods in which only one reaction was necessary to detect multiple target sequences that can be reliably used for the identification of specific lines of transgenic cows.

Key words: transgenic cow; multiplex PCR; hLF gene

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