转基因番木瓜事件特异性多重PCR检测方法的建立  

Establishment of Multiplex PCR Detection Method for Genetically Modified Papaya(Carica papaya)

作　　者：潘志文[1] 何荧 高洁儿 陈伟庭[1] 周峰[1] 姚涓[1] 姜大刚[1] PAN Zhi-Wen;HE Ying;GAO Jie-Er;CHEN Wei-Ting;ZHOU Feng;YAO Juan;JIANG Da-Gang(College of Life Sciences,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学生命科学学院,广州510642

出　　处：《农业生物技术学报》2021年第5期1016-1023,共8页Journal of Agricultural Biotechnology

基　　金：广东省农业科技创新及推广项目(2018LM7188)。

摘　　要：转基因抗病番木瓜(Carica papaya)为解决环斑病毒(Papaya ring spot virus,PRSV)对木瓜产业的危害做出了重要贡献。目前,主要的转基因番木瓜品系为’55-1’、’GM YK’、’华农1号’。为了对抗病转基因番木瓜进行更加有效的监管、保护消费者的知情权和选择权,需要建立更加高效、准确的转基因番木瓜检测方法。本研究分析全球主要的转基因番木瓜品系,针对番木瓜内标准基因Papain、筛选标记基因NPTⅡ和转基因抗病番木瓜品系’55-1’、’GM YK’、’华农1号’3个转化体的事件特异性序列,建立简便、高效的多重PCR检测方法。测试结果显示,本研究建立的多重PCR检测方法特异性强、灵敏度高,通过1次PCR和电泳检测即可判断待测样品是否为转基因番木瓜及其品系成分。本研究为转基因番木瓜成分检测和身份确认提供技术支持。Disease-resistant transgenic papaya(Carica papaya)has made great contribution in solving the problem of Papaya ring spot virus(PRSV).At present,the main transgenic papaya lines are’55-1’,’GM YK’and’Huanong No.1’.In order to supervise the disease-resistant transgenic papaya products in the market and protect consumers’right to know and right to choose,it is necessary to establish more efficient and accurate detection method for transgenic papaya.This study analyzed the major genetically modified(GM)papaya in the world and established a simple and efficient multiplex PCR detection method for specific sequences of the papaya endogenous reference gene Papain,screening marker gene NPTⅡ,event-specific sequences of GM papaya lines’55-1’,’GM YK’and’Huanong No.1’.The test results suggested that the established multiplex PCR detection method had high specificity and sensitivity.Samples could be judged whether they were transgenic papaya and which type of transgenic papaya were contained by only once PCR assay and electrophoresis.This study provides technical support for the composition detection and identity verification of transgenic papaya.

关 键 词：转基因番木瓜 多重PCR 检测方法 

分 类 号：S59[农业科学—作物学]