应用多重PCR技术快速筛查食品中的转基因成分

A Multiplex PCR Assay for the Rapid Screening of Genetically Modified Components in Food

DOI：10.3969/j.issn.1673-1689.2017.01.010

中文关键词: 转基因生物 调控元件 标记基因 目的基因 多重PCR 筛查

英文关键词: genetically modified organisms,regulatory element,marker gene,target gene,multiplex PCR,screening

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中文摘要:

以转基因植物中常见的3种筛选调控元件、2种标记基因和1种目的基因为检测靶标，建立了能在一次PCR扩增中同时检测6种转基因成分的多重PCR检测方法。结果表明，当多重PCR体系中CaMV35S启动子、FaMV35S启动子、NOS终止子、bar基因、NPTII基因、CP4-EPSPS基因的检测引物浓度分别为0.2、0.2、0.3、0.3、0.2、0.2 μmol/L，退火温度为60 ℃时扩增效果最佳，利用优化后的六重PCR方法可从转基因玉米、大豆、棉花、油菜等各类样品中精准检测出相应的转基因成分，方法的检测灵敏度可达到0.1%。该方法为食品中常见转基因成分的筛查提供了一种高效的手段，在转基因产品的检测工作中具有较高的应用价值。

英文摘要:

Based on the DNA sequences of CaMV35S promoter，FaMV35S promoter，NOS terminator，bar gene，NPTII gene and CP4-EPSPS gene，a multiplex PCR assay for simultaneously screening the above six exogenous elements widely introduced into genetically modified organisms was established. The result showed that the highly efficient multiplex PCR system was obtained when the final primer concentrations were 0.2，0.2，0.3，0.3，0.2 and 0.2 μmol/L separately for them. The method could allow an accurate detection of six target elements from genetically modified maize，soybean，cotton and rapeseed，with the limit of detection of 0.1%. In conclusion，this multiplex PCR assay provides a highly effective approach to the screening of common transgenic components in foods and has high application value in the GMO detection.

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