| 转基因玉米Bt176液相芯片检测方法的建立 |
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| 引用本文： | 程涛,王慧煜,黄新,余多慰,韩雪清.转基因玉米Bt176液相芯片检测方法的建立[J].生物技术通报,2012(4):158-164. |
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| 作者姓名： | 程涛 王慧煜 黄新 余多慰 韩雪清 |
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| 作者单位： | 1. 南京师范大学生命科学学院,南京,210046
2. 中国检验检疫科学研究院,北京,100029 |
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| 摘 要： | 为建立转基因玉米Bt176的液相芯片检测方法,根据已公布的转基因玉米Bt176外源插入基因CaMV35S启动子序列,外源基因3’端与玉米基因组DNA连接区序列,同时以玉米特异Zein内源基因序列为参照,利用Primer Premier5.0等软件设计特异性引物和探针。将探针与荧光编码微球偶联后,与PCR产物杂交反应,用液相芯片检测仪(Bio-plex 200)检测荧光信号。检测结果显示,该方法具有高特异性及灵敏度,各条探针之间无交叉反应,最低检测限可达0.01%。初步建立了检测转基因玉米Bt176的液相芯片技术,为其他转基因作物的快速高通量检测提供了借鉴和经验。
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| 关 键 词： | 转基因玉米Bt176 液相芯片 灵敏度 特异性 |
Development of Liquichip Detection Method for Detecting Genetically Modified Maize |
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| Cheng Tao
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Wang Huiyu
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Huang Xin
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Yu Duowei
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Han Xueqing.Development of Liquichip Detection Method for Detecting Genetically Modified Maize[J].Biotechnology Bulletin,2012(4):158-164. |
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| Authors: | Cheng Tao Wang Huiyu Huang Xin Yu Duowei Han Xueqing |
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| Institution: | 1College of Life Science,Nanjing Normal University,Nanjing 210046;2Chinese Academy of Inspection and Quarantine,Beijing 100029) |
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| Abstract: | This experiment was conducted to develop a liquichip detection technique for detecting genetically modified Bt176 maize.Based on the sequences of CaMV p35S,3’-transgenie integration sequence of Bt176 maize and maize endogenous reference gene Zein.Three primer pairs and three probes with high specificity were designed for each sequence using Primer Premier 5.0.The selected sets of beads were coupled by probes,than hybridized with PCR production.Finally,the detection step was done using the Bio-plex 200 System.The results indicated the method was highly specific and sensitive,without cross reactions among three probes,and the limit of detection was 0.01%.The liquichip detection method for detecting genetically modified Bt176 maize,could provide reference for other rapid high-throughput detection of genetically modified crops. |
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| Keywords: | Genetically modified Bt176 maize Liquichip Sensitivity Specificity |
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