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| 基于Special-Base模型检测转基因作物nos/plamid构建特异性 |
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兰青阔, 王永, 赵新, 陈锐, 朱珠, 李欧静, 余景会, 郭永泽, 程奕
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| (天津市农业科学院 中心实验室/农业部转基因产品成分监督检验测试中心, 天津 300381) |
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| 摘要: |
| 以转基因作物中常见的nos/plamid构建特异性序列为研究对象,通过同源性比较和焦磷酸测序dispensation order设计,建立Special-Base焦磷酸测序检测模型;利用转基因大豆Roundup Ready Soybean和转基因玉米Bt11、GA21、NK603 等4种转基因作物为试验材料,针对nos/plamid构建特异性序列设计引物,经复合PCR扩增及焦磷酸测序,验证Special-Base焦磷酸测序检测模型有效性。结果表明:本研究建立的Special-Base焦磷酸测序模型能检测出样品中单一或多种转基因作物成分,提高了转基因作物检测的精确性、检测通量和自动化程度。 |
| 关键词: 转基因作物(GMC) 复合PCR 焦磷酸测序 Special-Base nos/plamid |
| DOI：10.11841/j.issn.1007-4333.2012.02.002 |
| 投稿时间：2011-11-28 |
| 基金项目:科技部国际合作项目(2006DFA32380) |
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| Special-Base GMCs detection using multiple PCR and pyrosequencing |
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LAN Qing-kuo, WANG Yong, ZHAO Xin, CHEN Rui, ZHU Zhu, LI Ou-jing, YU Jing-hui, GUO Yong-ze, CHENG Yi
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| (Central Lab/Supervision and Test Center for Molecular Characteristics of Genetically Modified Organisms and Their Derivates of Ministry of Agriculture, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China) |
| Abstract: |
| Despite the various technologies in place for GMC test,a robostic method is highly demanded.The technology described here represents a breakthrough development in that direction.Aimming to nos/plamid gene of four GMCs(RRS,Bt11,GA21,NK603),by combining the multiple PCR with pyrosequencing,we developed a rapid assay for genotyping nos/plamid that relies on the identification of a single,type-specific ‘sentinel’ base.The prototype assay was developed to recognize the four GMCs and was capable of recognizing simultaneously multiple GMCs infections.The studies established a rapid,reliable and cost-effective assay. |
| Key words: GMCs multiple PCR pyrosequencing Special-Base nos/plamid |
