| 转基因番木瓜 55-1 转化事件定性 PCR 检测方法的建立 |
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| 引用本文： | 王恒波,肖乃衍,张华,陈平华.转基因番木瓜 55-1 转化事件定性 PCR 检测方法的建立[J].热带作物学报,2018,39(9):1751-1757. |
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| 作者姓名： | 王恒波 肖乃衍 张华 陈平华 |
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| 作者单位： | 1农业部甘蔗及制品监督检验测试中心
2福建农林大学国家甘蔗工程技术研究中心 |
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| 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。
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| 关 键 词： | 番木瓜 转基因 定性PCR 检测 |
Establishment of a Qualitative PCR Detection Method for Transgenic Papaya 55-1 |
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| WANG Hengbo,XIAO Naiyan,ZHANG Hua,CHEN Pinghua.Establishment of a Qualitative PCR Detection Method for Transgenic Papaya 55-1[J].Chinese Journal of Tropical Crops,2018,39(9):1751-1757. |
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| Authors: | WANG Hengbo XIAO Naiyan ZHANG Hua CHEN Pinghua |
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| Institution: | 1Quality Supervision, Inspection and Testing Center for Sugarcane and Derived Products, Ministry of Agriculture
2National Engineering Research Center of Sugarcane, Fujian Agriculture and Forestry University |
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| Abstract: | 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. |
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| Keywords: | papaya transgenic qualitative PCR detection |
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