| 抗虫转Bt基因水稻外源转基因成分环介导等温扩增技术检测方法的建立及应用 |
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| 引用本文： | 王永,兰青阔,赵新,朱珠,程奕.抗虫转Bt基因水稻外源转基因成分环介导等温扩增技术检测方法的建立及应用[J].天津农业科学,2012,18(1):7-10. |
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| 作者姓名： | 王永 兰青阔 赵新 朱珠 程奕 |
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| 作者单位： | 天津市农业科学院中心实验室,天津,300381 |
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| 基金项目： | 国际合作项目(2006DFA32380);天津市应用基础及前沿技术研究(08JCYBJC03700) |
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| 摘 要： | 以转基因Bt水稻品种科丰6号为主要研究对象,利用环介导等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP),针对cry1Ac晶体蛋白毒素基因的6个区域设计4条特异性引物,利用一种链置换DNA聚合酶(Bst DNA polymerase)在65℃保温30min,通过荧光显色完成对转基因检测工作。结果显示,该LAMP方法能够特异性检测cry1Ac基因,其最低定性检测限是传统PCR方法的10倍。本研究建立的针对转基因水稻cry1Ac基因的LAMP检测方法具有高度的特异性及稳定性,结果可靠,非常适合转基因抗虫水稻的快速检测。
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| 关 键 词： | 抗虫转Bt基因水稻 环介导等温扩增技术 检测 |
Development and Application of Loop-Mediated Isothermal Amplification for Detection of Transgenic Insect-resistant Rice with Bt Gene |
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| WANG Yong
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LAN Qing-kuo
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ZHAO Xin
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ZHU Zhu
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CHENG Yi.Development and Application of Loop-Mediated Isothermal Amplification for Detection of Transgenic Insect-resistant Rice with Bt Gene[J].Tianjin Agricultural Sciences,2012,18(1):7-10. |
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| Authors: | WANG Yong LAN Qing-kuo ZHAO Xin ZHU Zhu CHENG Yi |
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| Institution: | (Central Lab of Tianjin Academy of Agricultural Sciences,Tianjin 300381,China) |
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| Abstract: | To develop a loop-mediated isothermal amplification method for rapid detection of transgenic insect-resistant rice with Bt(Bacillus thuringiensis)gene.The detection assay is based on the loop-mediated isothermal amplification(LAMP) reaction.The cry1Ac gene was amplified by a set of four specially primers that recognize six distinct sequences of the target.The amplification can be obtained in 30 min by incubating all of the reagents in a single tube with Bst DNA polymerase at 65 °C.Results showed that sensitivity of the LAMP assay were 10 times higher on the detection limit for cry1Ac gene than conventional PCR assay.Our results clearly demonstrated that the LAMP-based assay was a sensitive and reliable means for the detection of transgenic insect-resistant rice with Bt gene. |
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| Keywords: | transgenic insect-resistant rice with Bt gene loop-mediated isothermal amplification(LAMP) detection |
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