摘要： 基于微滴式数字PCR（Droplet digital PCR，ddPCR）平台，以转基因玉米为例，建立了转基因作物（Genetically modified crops，GM crops）外源基因拷贝数分析方法，对待测样品进行了快速鉴定，并从T₀转基因玉米株系中鉴定出多个单拷贝单株。对该方法与实时荧光定量PCR（Quantitative real-time PCR，qRT-PCR）方法在分析结果的准确性方面进行了比较，从试验数据可以看出，2种检测方法的结果比较一致，单拷贝检测结果高度一致；但是ddPCR试验操作更加简便，试验结果可重复性强，试验数据更加准确可靠。研究表明，ddPCR方法是一种更加便捷、快速和准确的外源基因拷贝数分析新方法，基于其在准确性和灵敏度方面的显著优势，将会在转基因作物的外源基因拷贝数分析中得到广泛的应用。

关键词: 微滴式数字PCR, 转基因玉米, 外源基因, 拷贝数, 绝对定量

Abstract: The droplet digital PCR (ddPCR) method to evaluate the exogenous gene copy number in genetically modified crops (GM crops) as the example from genetically modified maize was developed,and some T₀ transgenic maize plants with single copy of EPSP gene were selected from all the samples to be detected.The results were also compared with those from quantitative real-time PCR (qRT-PCR),as could be seen from the experimental datas,the results from qRT-PCR(TaqMan) and ddPCR for EPSP were consistent,especially the single copy detection results of both in a high degree of consistency.And the ddPCR method was easy to operate,the result was repeatable and accurate.Therefore,the ddPCR would be well developed as a novel method for estimating transgenic copy number with high accuracy,which might be widely used in the exogenous genes copy number analysis in GM crops in the near future.

Key words: Droplet digital PCR, Genetically modified maize, Exogenous gene, Copy number, Absolute quantification

中图分类号: 

• S513.03