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转基因番木瓜55-1品系的环介导等温扩增检测方法的建立
引用本文:夏启玉,李美英,肖苏生,贺萍萍,杨小亮,张雨良,郭安平.转基因番木瓜55-1品系的环介导等温扩增检测方法的建立[J].热带作物学报,2018,39(2):308-316.
作者姓名:夏启玉  李美英  肖苏生  贺萍萍  杨小亮  张雨良  郭安平
作者单位:1. 中国热带农业科学院热带生物技术研究所 2农业部热带作物生物学与遗传资源利用重点实验室 3农业部转基因植物及植物用微生物环境安全监督检验测试中心
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。 

关 键 词:转基因番木瓜55-1  环介导等温扩增  特异性检测  

Development of a Loop-mediated Isothermal Amplification Method for Detection of Genetically Modified Papaya Line 55-1
XIA Qiyu,LI Meiying,XIAO Susheng,HE Pingping,YANG Xiaoliang,ZHANG Yuliang,GUO Anping.Development of a Loop-mediated Isothermal Amplification Method for Detection of Genetically Modified Papaya Line 55-1[J].Chinese Journal of Tropical Crops,2018,39(2):308-316.
Authors:XIA Qiyu  LI Meiying  XIAO Susheng  HE Pingping  YANG Xiaoliang  ZHANG Yuliang  GUO Anping
Institution:1 Institute of Tropical Bioscience and Biotechnology , Chinese Academy of Tropical Agriculture Sciences 2Key Laboratory of Biology and Genetic Resources of Tropical Crops , Ministry of Agriculture, Haikou, Hainan 571101, China;  3 Environmental Safety Supervision and Inspection Center for Genetically Modified Plants and Microorganism used in Plants, Ministry of Agriculture, Haikou, Hainan 571101, China
Abstract: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 transgenic papaya.
Keywords:genetically    modified    papaya    line    55-1  loop    mediated    isothermal    amplification  specific    detection  
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