| 一种检测转Bt基因抗虫棉新棉33B和GK-12的PCR方法 |
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| 引用本文： | 王奕海,谢家建,张永军,王锡锋,彭于发.一种检测转Bt基因抗虫棉新棉33B和GK-12的PCR方法[J].农业生物技术学报,2009,17(5):914-919. |
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| 作者姓名： | 王奕海 谢家建 张永军 王锡锋 彭于发 |
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| 作者单位： | 中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京,100193 |
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| 基金项目： | 国家重大基础研究发展规划(973)项目 |
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| 摘 要： | 测定了转基因抗虫棉新棉33B和GK-12的Bt基因表达盒的序列，发现它们在Bt基因和Bt基因与终止子连接区的序列存在差异，而在启动子与Bt基因连接区的序列完全一致。基于这种结构上的差异，设计3条特异性引物MG-P1、MG-P2和MG-P3，建立了检测这两个转基因抗虫棉的双重PCR方法。采用建立的方法，检测了40个转基因棉花样品，其中，只含有新棉33B的Bt基因结构的样品数为32个；只含有GK-12的Bt基因结构的样品数为6个；同时含有两者结构的样品数为2个。
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| 关 键 词： | 转基因棉花 Bt基因 检测 双重PCR |
| 收稿时间： | 2008-12-15 |
| 修稿时间： | 2009-3-25 |
A PCR Method to Detect Different Bt Gene Expression Cassettes in Transgenic Bt Cotton |
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| Abstract: | The Bt gene expression cassettes of GM cotton 33B and GK-12 were sequenced and analyzed. Differences were found both in the Bt gene and the junction region of Bt gene/terminators except the junction region of CaMV 35S promoter/Bt gene. In order to detect the two transgenic cotton lines, three specific primers, MG-P1, MG-P2 and MG-P3, were designed and the duplex PCR detection method was established based on the differences. Forty transgenic cotton samples were tested by this duplex PCR method, thirty-two samples were identified to contain 33B Bt gene structure, six samples were identified to have GK-12 Bt gene structure, and two samples were identified to have both 33B and GK-12 Bt gene structures. |
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