根据MON89034玉米的5’端和3’端边界序列分别设计1组转化体特异性的巢式PCR引物，采用中途进退式PCR策略建立MON89034玉米的转化体特异性检测方法，扩增产物分别为491 bp和188 bp。以转基因玉米MON89034及8种其他转基因作物为材料，证明此方法对MON89034玉米具有高度特异性。灵敏度测试结果表明，此方法的相对检出限达到0.01%，绝对检出限为4个单倍体基因组拷贝数，比普通PCR提高了5倍。建立的单管巢式和半巢式PCR方法可准确、高效地检测转基因玉米MON89034及其产品。

The two sets of event-specific nested PCR primers were designed based on the 5' - and 3'- flanking sequence of the exogenous integrant of GM maize MON89034 respectively, with the aim of developing a drop-in drop-out PCR assay for the event-specific detection of GM maize MON89034, resulting in two amplification fragment of 491 bp and 188 bp in length respectively. This assay has been successfully applied to distinguish GM maize MON89034 from GM maize MON810, MON863, Bt11, Bt176, MON88017, GM roundup ready soybean and GM canola GT73 with high specificity. Sensitivity test results showed that the relative limit of detection of this method reaches 0.01%, equivalent to the absolute sensitivity of 4 copies of haploid genome, which was 5 times more than conventional PCR. Consequently, the single tube nested and semi-nested PCR assay established in this study can be used to accurately and efficiently detect GM maize MON89034 and its processed products.

国家转基因生物新品种培育科技重大专项课题（2016ZX08012-001）、吉林省农业科技创新工程项目（2013）