转基因苜蓿草J163品系特异性LAMP检测 方法的建立

Establishing an event-specific loop–mediated isothermal amplification detec-tion method for genetically modified alfalfa events J163

中文关键词:  等温扩增  转基因苜蓿J163  品系特异性

英文关键词:loop-mediated isothermal amplification  genetically modified alfalfa J163  event-specific

中文摘要:

      目的 建立针对我国农业部未颁发农业转基因生物安全证书的转基因苜蓿草品系J163品系特异性等温扩增(loop-mediated isothermal amplification, LAMP)检测方法。方法 根据转基因苜蓿草品系J163 5’端外源插入片段与苜蓿草基因组DNA之间的邻接区序列设计6条引物, 建立了转基因苜蓿草J163品系特异性LAMP检测方法, 并对本方法的特异性、灵敏度进行了测定。结果 建立的检测方法特异于转基因苜蓿草J163成分检测, 检测最低DNA浓度为(1imit of detection, LOD)为16 pg, 相当于10拷贝转基因苜蓿草J163基因组DNA。结论 本研究建立的转基因苜蓿草J163品系特异性LAMP检测方法特异性好, 灵敏度高, 能够快速、准确、稳定地对转基因苜蓿草J163成分进行检测分析。

英文摘要:

      Objective To establish a loop–mediated isothermal amplification(LAMP) detection method for genetically modified (GM) Roundup Ready Alfalfa Events J163, which has not been authorized by the Ministry of Agriculture of China. Methods The specific primer pairs based on the 5’junction sequence spanning the alfalfa DNA and inserted fragment of J163 were designed and then the LAMP detection system was established. The specificity and sensitivity were analyzed. Results The LAMP method was specific for GM alfalfa J163 detection, the limit of detection(LOD) were 16 pg J163 genomic DNA or 10 copies of alfalfa J163 haploid genomic DNA. Conclusion The established event-specific LAMP method for GM alfalfa J163 detection had a high specificity and good sensitivity, and was suitable for quantification of J163 samples quickly and accurately.