抗虫玉米MON89034转化体特异性PCR检测技术研究

Event-specific Qualitative PCR Method for Detection of Maize Line MON89034

根据抗虫玉米MON89034外源插入片段5’端与植物基因组连接区序列设计特异性引物,并进行PCR扩增,预期产物大小为455 bp。以zSSIIb基因作为内标准基因,建立了转基因玉米MON89034转化体特异性定性PCR检测方法。对该方法进行重现性、特异性和灵敏度测试,结果表明:该方法能够特异性检测出MON89034转化体,以100 ngDNA为模板,该方法的检测灵敏度达到0.1%,约为40个起始模板拷贝。复合PCR检测结果还表明,在同一PCR反应管中可实现对zSSIIb基因和MON89034的同时检测。

The specific primers were designed based on the 5’-junction sequences of the exogenous integrant of maize MON89034,and amplification products of 455 bp were obtained.The ruggedness,specificity and sensitivity of this method were tested.The results showed that MON89034 could be distinguished specifically from other GM and non-GMcrops by using this method,and the limit of detection was up to 0.1%.In addition,a multiplexed PCR reac-tion was developed by which specific fragment of MON89034 and one fragment of m...