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应用单管半巢式PCR技术筛查转基因食品
引用本文:闫 伟,徐桢惠,龙丽坤,李葱葱,李飞武. 应用单管半巢式PCR技术筛查转基因食品[J]. 食品科学, 2015, 36(2): 194-197. DOI: 10.7506/spkx1002-6630-201502037
作者姓名:闫 伟  徐桢惠  龙丽坤  李葱葱  李飞武
作者单位:1.吉林省农业科学院农业生物技术研究所,吉林 长春 130033;2.哈尔滨师范大学生命科学与技术学院,黑龙江 哈尔滨 150025
基金项目:国家转基因生物新品种培育科技重大专项(2014ZX08012-001);吉林省农业科技创新工程项目(2013)
摘    要:建立转基因作物中常见的两种外源调控元件的单管半巢式聚合酶链式反应(polymerase chain reaction,PCR)测方法,为转基因食品的筛选检测提供一种快速、精确的方法。针对6 种转基因作物中CaMV35S启动子和NOS终止子的一致性核苷酸序列,分别设计巢式PCR的内、外引物,在测试不同引物组合的扩增效率基础上,建立CaMV35S启动子和NOS终止子的单管半巢式PCR方法。结果表明,上述方法对两种转基因成分具有良好的特异性,检测灵敏度分别为0.01%和0.05%,显著优于常规PCR方法。该方法具有便捷、准确、灵敏等特点,适合筛查食品中的转基因成分。

关 键 词:转基因食品  单管半巢式PCR  筛选检测  

Screening of Genetically Modified Foods by Single-Tube Semi-Nested PCR
YAN Wei,XU Zhenhui,LONG Likun,LI Congcong,LI Feiwu. Screening of Genetically Modified Foods by Single-Tube Semi-Nested PCR[J]. Food Science, 2015, 36(2): 194-197. DOI: 10.7506/spkx1002-6630-201502037
Authors:YAN Wei  XU Zhenhui  LONG Likun  LI Congcong  LI Feiwu
Affiliation:1. Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, Changchun 130033, China;2. College of Life Science and Technology, Harbin Normal University, Harbin 150025, China
Abstract:A single-tube semi-nested PCR method for common exogenous regulatory elements of transgenic crops was
established to provide a fast and accurate assay for screening genetically modified foods. Two sets of element-specific nested
PCR primers were designed based on the consensus sequence of the exogenous elements in six different transgenic crops,
respectively. A single-tube semi-nested PCR assay for CaMV35S promoter and NOS terminator was developed by testing
the amplification efficiency of different combinations of primers. This assay has been successfully applied to distinguish
GM crops with CaMV35S promoter or NOS terminator from others with high specificity. Sensitivity tests showed that the
relative limit of detection for CaMV35S promoter and NOS terminator were 0.01% and 0.05%, respectively, significantly
better than those of conventional PCR. Consequently, the single-tube semi-nested PCR assay is convenient, accurate,
sensitive and suitable for screening genetically modified foods.
Keywords:genetically modified foods  single-tube semi-nested PCR  screening and detection  
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