摘要
根据55-1的品系特异性序列设计了5套LAMP引物,筛选扩增效率高的引物。对筛选到的LAMP引物的反应温度和反应体系进行了优化,并进行了特异性、灵敏度和稳定性的实验。结果表明,该套LAMP引物的特异性和稳定性良好,灵敏度为0.05%,最快能在30 min内得到检测结果。建立的LAMP检测方法能有效地检测转基因番木瓜55-1品系,既简化了检测步骤,又缩短了检测时间,为转基因番木瓜的快速检测提供了技术支撑。
The purpose of the study is to develop an loop-mediated isothermal amplification（LAMP）technique for the detection of genetically modified papaya line 55-1. Five sets of LAMP primers were designed according to the specific sequences of genetically modified papaya line 55-1, and primers with high amplification efficiency were screened. The LAMP reaction temperature and reaction system of the set of primers were optimized, and the specificity, sensitivity and stability of the LAMP method were determined. The results indicated that the set of LAMP primers could detect genetically modified papaya line 55-1 specifically and stably, with a sensitivity of 0.05% and a detection result within 30 min. Therefore, the LAMP method can effectively detect the genetically modified papaya line 55-1, which not only simplifies the detection steps, but also shortens the detection time, and provides technical support for the rapid detection of transgenie papaya.
作者
夏启玉
李美英
肖苏生
贺萍萍
杨小亮
张雨良
郭安平
XIA Qiyul,2, LI Meiying1,2, XIAO Susheng1,2, HE Pingping1,2, YANG Xiaoliang1,2, ZHANG Yuliang1,2, GUO Anping1,2(Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences~Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Haikou, Hainan 571101, China Environmental Safety Supervision and Inspection Center for Genetically Modified Plants and Microorganism used in Plants, Ministry of Agriculture, Haikou, Hainan 571101, Chin)
出处
《热带作物学报》
CSCD
北大核心
2018年第2期308-316,共9页
Chinese Journal of Tropical Crops
基金
海南省重点研发计划项目(No.ZDYF2017017)
中央级公益性科研院所基本科研业务费专项(No.1630052016019
No.1630052016001)
中国热带农业科学院基本科研业务费专项资金(No.17CXTD-24)
关键词
转基因番木瓜55-1
环介导等温扩增
特异性检测
genetically modified papaya line 55-1
loop mediated isothermal amplification
specific detection
