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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 according to the 5 ’end of exogenous insert of insect-resistant maize MON89034 and the plant genomic junction region, and PCR amplification was performed. The expected product size was 455 bp. The zSSIIb gene was used as an internal standard to establish a specific qualitative PCR method for transgene expression analysis of MON89034 in maize. The reproducibility, specificity and sensitivity of this method were tested. The results showed that this method could detect MON89034 transformant specifically. The detection sensitivity of this method was 0.1% with 100 ng DNA as template, and about 40 initial templates copy. The multiplex PCR results also showed that simultaneous detection of the zSSIIb gene and MON89034 was achieved in the same PCR tube.