利用实时荧光PCR快速筛查玉米中转基因成分 |
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引用本文:张海波, 刘 冰, 杨娟妮, 张 田, 张 英.利用实时荧光PCR快速筛查玉米中转基因成分[J].西北农业学报,2017,26(6):840~848 |
DOI:10.7606/j.issn.1004-1389.2017.06.006 |
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中文摘要:玉米是拥有转基因转化体数量最多的作物,多样的转化体增加了玉米中转基因成分筛查的难度。为了建立玉米中转基因成分的快速筛查方法,利用Taqman探针实时荧光PCR方法,通过对玉米样品中CaMV35S启动子(P-35S)和NOS终止子(T-NOS)2个元件组合的检测,完成对现有玉米转化体的筛查。建立的Taqman探针实时荧光PCR方法灵敏度可达到0.1%,检测时间较普通PCR方法缩短1 h以上。同时提出了转基因玉米转化体鉴定路线图。该筛查方法可对玉米中的转基因成分进行快速、高灵敏度的筛查。 |
中文关键词:转基因玉米 Taqman探针实时荧光PCR 快速筛查 CaMV35S启动子 |
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Rapid Screening of Genetically Modified Maize Based on Taqman Real-time Fluorescence PCR Method |
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Abstract:Maize is the crop of having the largest amount of genetically modified (GM) events. Varied GM events make it difficult to screen GM ingredients in maize. In order to establish a rapid screening method for GM maize, the combination of CaMV35S promoter (P-35S) and NOS terminator (T-NOS) was used to screen all the commercial GM maize events based on Taqman real-time fluorescence PCR. The sensitivity of the method was of 0.1%.The time cost in Taqman real-time fluorescence PCR was one hour less than conventional PCR.In addition, the identification pathway of genetically modified maize event was presented in this article. In conclusion, this Taqman real-time fluorescence PCR method can be used for rapid and sensitive screening of GM maize events. |
keywords:Genetically modified maize Taqman Real-time PCR Method Rapid Screening CaMV35S Promoter |
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