论文部分内容阅读
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。
A set of transformant-specific nested PCR primers was designed according to the 5 ’and 3’ border sequences of MON89034 maize. The transformant specific detection method of MON89034 maize was established by the forward and backward PCR strategy. The amplification products were 491 bp and 188 bp. Using transgenic corn MON89034 and eight other transgenic crops as materials, this method proved to be highly specific to MON89034 maize. Sensitivity test results showed that the relative detection limit of this method was 0.01%, and the absolute detection limit was 4 haploid genome copy number, which was 5 times higher than that of normal PCR. The established single-tube nested and semi-nested PCR methods can accurately and efficiently detect transgenic corn MON89034 and its products.