| 实时荧光定量PCR法检测抗虫棉Bt基因拷贝数方法的建立 |
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| 引用本文： | 闫喜中,张锐,孟志刚,孙国清,周涛,郭三堆. 实时荧光定量PCR法检测抗虫棉Bt基因拷贝数方法的建立[J]. 安徽农业科学, 2008, 36(26) |
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| 作者姓名： | 闫喜中 张锐 孟志刚 孙国清 周涛 郭三堆 |
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| 作者单位： | 中国农业科学院生物技术研究所/国家农作物基因资源与基因改良重大科学工程 |
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| 摘 要： | [目的]建立并完善Taqman实时荧光定量PCR检测转基因抗虫棉Bt基因的方法。[方法]通过Bt基因序列,利用软件Primer5.0设计引物和探针,然后利用10倍系列稀释含有目的基因的质粒pG4AB,进行实时荧光定量PCR反应,制作标准曲线。[结果]抗虫基因拷贝数与Ct值的关系为Ct=-3.549 632X+43.783 512,相关系数R2为0.997 867。[结论]实时荧光定量PCR检测转基因抗虫棉Bt基因的方法具有特异性好、灵敏度高的特点。
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| 关 键 词： | Bt基因 实时荧光定量PCR 标准曲线 |
Establishing a Real-time Quantitative PCR Assay for Detecting the Copy Number of Bt Gene in Transgenic Insect-resistant Cotton |
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| YAN Xi-zhong et al. Establishing a Real-time Quantitative PCR Assay for Detecting the Copy Number of Bt Gene in Transgenic Insect-resistant Cotton[J]. Journal of Anhui Agricultural Sciences, 2008, 36(26) |
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| Authors: | YAN Xi-zhong et al |
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| Abstract: | [Objective] The aim of this paper was to develop real-time quantitative PCR assay for detecting Bt gene in transgenic insect-resistant cotton.[Method] The primers and probes of Bt gene were designed using the software primer 5.0.The standard curve was prepared based on the linear relationship between the amount of input plasmid DNA(X) and cycle threshold(Ct).[Result] For Bt gene,Ct=-3.549 632X 43.783 512,and correlation coefficient R2 was 0.997 867.[Conclusion]The real-time quantitive PCR method could be used as an effective detection and quantification method for Bt gene and it is amenable to high-throughout assay for its specificity,sensitivity and rapidity. |
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| Keywords: | Bt gene Real-time quantitative PCR Standard curve |
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