LAMP实时浊度法检测转基因植物NPTⅡ基因  

Development of a Real-time Turbidimeter-based LAMP Method for Detection of NPT Ⅱ Gene in the Genetically Modified Plant

作　　者：邝筱珊[1] 胡松楠[1] 王小玉[1] 唐食明[1] 成晓维[1] 冯家望[1] 

机构地区：[1]珠海出入境检验检疫局技术中心,珠海519015

出　　处：《生物技术通报》2014年第3期60-64,共5页Biotechnology Bulletin

基　　金：国家质检总局科技计划项目(2011IK255)

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(LAMP)实时浊度仪 新霉素-3'-磷酸转移酶基因 转基因 检测 

分 类 号：Q943[生物学—植物学]