转基因玉米LY038转化事件的特异性检测

摘要

Genetically modified maize with high lysine LY038 has been widely applied in animal feed, but so far there has been no report about the method of amplification for flanking sequence of LY038 and its event-specific qualitative PCR detection. In this study, the 5'- flanking sequence of integrated gene-construct of genetically modified maize LY038 was characterized by modified adapter-linked PCR (M-AL-PCR). According to the 5'flanking sequence, the event-specific primers were designed, and the event-specific qualitative detection of genetically modified plant LY038 was established to produce a 175 bp product. With genetically modified maize (Zea mays L.) LY038, MIR604, Bt176, Btll, MON810, MON863, GA21, NK603, non-genetically modified maize, genetically modified rice (Oryza sativa L.) CrylC* and Cry2A*, genetically modified soybean (Glycine max) Roundup Ready and genetically modified oilseed rape (Brassica campestris L.) GT73 as materials, this method developed in this work proved to be highly specific and sensitive, and the limit of detection was 0.1％. It is suggested that the developed qualitative detection system can accurately, fast and efficiently identify the genetically modified maize LY038 and its derivates.%转基因高赖氨酸玉米LY038在畜禽饲料中已得到广泛应用,但目前还没有关于其侧翼序列扩增及转化事件特异性定性PCR检测方法方面的报道.本研究采用修饰接头连接PCR(modified adapter-linked PCR,M-AL-PCR)技术获得了转基因玉米LY038的外源基因与玉米基因组之间的5′端侧翼序列.据此侧翼 序列设计其转化事件特异性引物,建立了转基因玉米LY038转化事件特异性定性检测方法,扩增片段175bp.以转基因玉米(Zea mays L.)LY038、MIR604、Bt176、Bt11、MON810、MON863、GA21、NK603、非转基因玉米、转基因水稻(Oryza sativa L.)Cry1C*、Cry2A*、转基因大豆(Glycine max)Roundup Ready和转基因油菜(Brassica campestris L.)GT73为材料,验证该方法具有高特异性及灵敏度,最低检测限约为0.1%.研究结果提示,该定性检测体系可准确、快速、高效的检测转基因玉米LY038及其产品.