转基因番木瓜阳性质粒分子的构建和应用  

Construction and Application of Positive Plasmid Molecular for Genetically Modified Papaya Detection

作　　者：陈伟庭[1] 高洁儿 潘志文[1] 周峰[1] 王声斌[1] 姜大刚[1] 姚涓[1] Chen Weiting;Gao Jieer;Pan Zhiwen;Zhou Feng;Wang Shengbin;Jiang Dagang;Yao Juan(Inspection and Testing Center for Ecological and Environmental Risk Assessment of Plant and Plant-Related Microorganism(Guangzhou),Ministry of Agriculture and Rural Affairs,College of Life Sciences,South China Agricultural University,Guangzhou,510642)

机构地区：[1]华南农业大学生命科学学院,农业农村部植物及植物用微生物生态环境安全监督检验测试中心(广州),广州510642

出　　处：《分子植物育种》2022年第18期6041-6048,共8页Molecular Plant Breeding

基　　金：广东省农业科技创新项目(2018KM7188)资助。

摘　　要：转基因抗病番木瓜已经获得了全球多个国家的商业化应用和进口批准。为了解决转基因番木瓜检测工作中存在阳性对照不易获取的问题,本研究通过基因数据库(美国国家生物信息中心,NCBI),筛选番木瓜内标准基因Papain和Chymopapain基因以及已经报道的番木瓜转化体‘华农1号’、‘55-1’和‘GM YK’的特异性序列,构建了转基因番木瓜阳性质粒分子,并对其特异性、灵敏度以及适用性进行了验证分析,结果表明:该阳性质粒分子包含的5个外源片段的扩增产物条带清晰、特异性好,检测灵敏度高,可以稳定地检测出100个拷贝的质粒分子;同时对质粒分子的适用性进行了测试,结果显示,构建的质粒分子可以应用于番木瓜组织和加工品的转基因检测中。由此表明本研究构建的转基因番木瓜阳性质粒分子可以作为阳性对照应用于转基因番木瓜的特异性检测。Disease-resistant transgenic papaya has been approved for commercial application and import in many countries around the world.To solving the difficulty of obtaining the positive control for the detection of transgenic papaya,we constructed a transgenic papaya positive plasmid molecular through searching the database of the National Center for Biotechnology Information in this study.The plasmid molecular contained two papaya endogenous reference genes of Papain and Chymopapain,the reported event-specific flanking sequence of transgenic papaya’Huanong No.1’,’55-1’and’GM YK’.The specificity testing result showed that the amplification products of five fragments had clear and specific bands.100 molecular copies can be stably detected in the sensitivity testing.Its’applicability experiments testing showed that it can be used in papaya tissues and processing products detecting.These results indicated that the papaya positive plasmid molecular can be recommended in the detection of transgenic papaya detection.

关 键 词：转基因番木瓜 阳性质粒分子 检测 事件特异性 

分 类 号：S667.9[农业科学—果树学]