| 转基因大豆MON89788转化体特异性定性PCR检测 |
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| 引用本文： | 李飞武,李葱葱,董立明,邢珍娟,张明.转基因大豆MON89788转化体特异性定性PCR检测[J].安徽农业科学,2010,38(13):6679-6682. |
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| 作者姓名： | 李飞武 李葱葱 董立明 邢珍娟 张明 |
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| 作者单位： | 吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033 |
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| 基金项目： | 国家转基因生物新品种培育重大专项 |
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| 摘 要： | 目的]建立MON89788大豆转化体特异性定性PCR检测方法。方法]利用TAIL-PCR技术分离MON89788大豆的3′端旁侧序列,据此序列设计特异性引物进行PCR检测,并对该方法的特异性和灵敏度进行测试。结果]获得了1142bp的3′端旁侧序列;依据该序列建立的PCR检测方法能特异性从MON89788大豆中扩增出170bp的产物,检测灵敏度达到0.05%,约为40个起始模板拷贝。结论]该研究建立的方法特异性强、灵敏度高,可适用于MON89788大豆检测。
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| 关 键 词： | 转基因大豆 MON89788转化体 定性PCR |
Establishment of Event-specific Qualitative PCR Method for Transgenic Soybean MON89788 |
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| Institution: | LI Fei-wu et al (Jilin Academy of Agricultural Sciences,National Center for Environmental Safety Inspection of Transgenic Plants,MOA,Changchun,Jilin 130033) |
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| Abstract: | Objective] The aim was to establish an event-specific qualitative PCR method for transgenic soybean MON89788. Method] Firstly,the 3’-junction sequence between host plant DNA and integrated DNA of transgenic MON89788 soybean was isolated using thermal asymmetric interlaced-PCR (TAIL-PCR),and the specific PCR primers were designed based on the 3’-junction sequence. Secondly,the specificity and sensitivity of the qualitative PCR detection methods employing these primers were tested. Result] 1142-bp 3’-junction sequence was obtained. According to the sequence,event-specific qualitative PCR method was established,amplifying a 170-bp product specifically from MON89788 event,and the limit of detection was 0.05%,approximately 40 initial template copies. Conclusion] The method was highly specific,sensitive,and suitable for detection of MON89788 event. |
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| Keywords: | Transgenic soybean MON89788 event Qualitative PCR |
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