转基因番木瓜55-1转化事件定性PCR检测方法的建立  

Establishment of a Qualitative PCR Detection Method for Transgenic Papaya 55-1

作　　者：王恒波[1] 肖乃衍 张华[1] 陈平华[1] WANG Hengbo;XIAO Naiyan;ZHANG Hua;CHEN Pinghua(Quality Supervision,Inspection and Testing Center for Sugarcane and Derived Products,Ministry of Agriculture/National Engineer- ing Research Center of Sugarcane,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]农业部甘蔗及制品监督检验测试中心/福建农林大学国家甘蔗工程技术研究中心,福建福州350002

出　　处：《热带作物学报》2018年第9期1751-1757,共7页Chinese Journal of Tropical Crops

基　　金：国家标准委设立"转基因植物及其产品成分检测抗病毒转CP基因番木瓜定性PCR方法(No.20100644-T-326);抗病毒番木瓜‘55-1 63-1’及其衍生产品定性PCR方法课题(No.20100643-T-326)";现代农业产业技术体系建设专项资金(No.nycytx-24)

摘　　要：为了防止国外商业化生产的转基因番木瓜流入国内市场,建立转基因番木瓜55-1转化事件定性PCR检测方法,对于保护国内消费者知情权意义重大。本研究以转基因抗环斑病毒番木瓜55-1为研究材料,利用外源基因和番木瓜基因组序列设计了9对特异性检测引物,通过特异性引物筛选、熔解曲线分析、退火温度优化、特异性验证、灵敏度分析及检测限验证,建立转基因番木瓜55-1转化事件定性PCR检测方法。结果表明:本研究筛选出的检测引物,可特异的检出转基因番木瓜55-1转化事件,引物的检测灵敏度达到了0.1%的标准,高于欧盟0.9%的检测要求,完全可满足转基因检测标识制度的顺利实施。Transgenic papaya 55-1, which has been marketed in America for many years, is a genetically modified organism not currently approved for food in China. The paper aimed to establish a qualitative PCR detection method for transgenic papaya line 55-1 and its derivative products for preventing foreign transgenic papaya products into the Chinese market. Based on the exogenous gene and the papaya genome sequence, nine pairs of specific primers were designed with transgenic papaya 55-1 as the research material, using specific primer screening, melting curve analysis and optimization of annealing temperature, specific validation, sensitivity analysis and detection limit verification. The method of transgenic papaya 55-1 and it＇s derivates was established with qualitative PCR detection. The resulted showed that the screened primers, which could specialize to detect the transgenic papaya line 55-1, could reach 0.1% and the detection sensitivity of primers was higher than the requirements of the detection of EU 0.9%. The method of this paper would provide a scientific and reasonable experiment reference for validating the composition of genetic modified organisms in the future in China.

关 键 词：番木瓜 转基因 定性PCR 检测 

分 类 号：S432.42[农业科学—植物病理学]