使用多重聚合酶链式反应结合寡核苷酸微阵列对七种转基因大豆和玉米进行特定事件检测。

Event-specific detection of seven genetically modified soybean and maizes using multiplex-PCR coupled with oligonucleotide microarray.

作者信息

Xu Jia, Zhu Shuifang, Miao Haizhen, Huang Wensheng, Qiu Minyan, Huang Yan, Fu Xuping, Li Yao

机构信息

State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai 200433, P.R. China.

出版信息

J Agric Food Chem. 2007 Jul 11;55(14):5575-9. doi: 10.1021/jf070433m. Epub 2007 Jun 9.

DOI:10.1021/jf070433m

PMID:17559227

Abstract

With the increasing development of genetically modified organism (GMO) detection techniques, the polymerase chain reaction (PCR) technique has been the mainstay for GMO detection. An oligonucleotide microarray is a glass chip to the surface of which an array of oligonucleotides was fixed as spots, each containing numerous copies of a sequence-specific probe that is complementary to a gene of interest. So it is used to detect ten or more targets synchronously. In this research, an event-specific detection strategy based on the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene is being developed for its high specificity using multiplex-PCR together with oligonucleotide microarray. A commercial GM soybean (GTS 40-3-2) and six GM maize events (MON810, MON863, Bt176, Bt11, GA21, and T25) were detected by this method. The results indicate that it is a suitable method for the identification of these GM soybean and maizes.

摘要

随着转基因生物（GMO）检测技术的不断发展，聚合酶链反应（PCR）技术一直是转基因生物检测的主要手段。寡核苷酸微阵列是一种玻璃芯片，其表面固定有一系列寡核苷酸作为斑点，每个斑点包含大量与感兴趣基因互补的序列特异性探针拷贝。因此，它可用于同步检测十个或更多目标。在本研究中，基于宿主植物基因组DNA与整合基因之间独特且特异的整合连接序列，正在开发一种事件特异性检测策略，该策略利用多重PCR和寡核苷酸微阵列，具有高特异性。通过该方法检测了一种商业化转基因大豆（GTS 40-3-2）和六种转基因玉米事件（MON810、MON863、Bt176、Bt11、GA21和T25）。结果表明，该方法适用于这些转基因大豆和玉米的鉴定。