作　　者： ; ; ; ; ; ;

机构地区： 珠海出入境检验检疫局

出　　处： 《生物技术通报》 2014年第3期60-64,共5页

摘　　要： 根据转基因植物常用的选择标记基因NPTⅡ(HE582394.1)的序列,设计6条特异性LAMP引物,利用实时浊度仪对反应体系中扩增产物的实时监控筛选出最佳引物,最终建立转基因植物NPTⅡ基因的LAMP检测方法,并对该方法的特异性、灵敏度、稳定性进行了评价。结果表明,该方法能够特异性检出含有NPTⅡ基因的植物及其加工产品,特异性高,灵敏度达0.5%。对转基因玉米MON863(含NPTⅡ基因)含量为10%、1%、0.5%、0%(W/W)的样品DNA分别进行扩增,其稳定性好,无假阳性和假阴性。所建立的LAMP方法适用于特异性筛选检测含NPTⅡ基因的植物及其加工样品。 According to NPTⅡ gene（HE582394.1）, the common selective marker gene of genetically modified plant（GMP）, six specific primers were designed. A loop-mediated isothermal amplification（LAMP）method for screening GMP was established. Real-time monitoring of the LAMP reaction was achieved by a turbidimeter. Specificity, sensitivity, stability and repeatability of this method were tested. The results showed that the method can specifically detect NPTⅡ gene, and the detection sensitivity of the method was 0.5%. With the genetically modified maize MON863（NPTⅡ positive）samples of 10%, 1%, 0.5%, 0%（W/W）as templates, stability and repeatability testing was conducted, and false negative rate was 0. The results showed that the LAMP method is suitable for specifically detecting NPTⅡ gene in GMP.

关 键 词： 环节导等温扩增法 实时浊度仪 新霉素 磷酸转移酶基因 转基因 检测

领　　域： [生物学]