| 转基因小麦的定性PCR筛选检测技术 |
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| 引用本文： | 栾凤侠,张洪祥,白月.转基因小麦的定性PCR筛选检测技术[J].麦类作物学报,2007,27(3):378-381. |
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| 作者姓名： | 栾凤侠 张洪祥 白月 |
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| 作者单位： | 黑龙江出入境检验检疫局,黑龙江哈尔滨,150001 |
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| 基金项目： | 国家质量监督检验检疫总局科研项目
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科技部社会公益研究专项重点项目子课题 |
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| 摘 要： | 为了建立转基因小麦的PCR检测方法标准,以B73、B72、B102三个被转入外源高分子量谷蛋白亚基(HMW-GS)基因的转基因小麦品系为材料,对目前国内外转基因小麦中通用的标记基因bar和uidA、NOS终止子和Ubiquitin启动子进行了定性PCR的筛选检测,在国内外首次找到了小麦中特有的内参照基因麦谷醇溶蛋白基因(GAG56D),设计并合成了麦谷醇溶蛋白基因(GAG56D)和Ubiquitin启动子的引物序列,并对PCR反应条件进行了优化,PCR产物经过琼脂糖凝胶电泳分析后,可以检测到预期大小的目的片段.同时对PCR产物用实时荧光定量PCR进行了确证实验,并得到预期的结果.定性PCR最低检测灵敏度为0.5%(w/w).建立的转基因小麦定性PCR筛选检测方法具有通用性.
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| 关 键 词： | 小麦 转基因 PCR 定性检测 转基因 小麦 筛选检测 检测技术 Transgenic Wheat Components Modified Detecting 通用性 检测方法 检测灵敏度 结果 确证实验 实时荧光定量 大小 预期 凝胶电泳分析 琼脂糖 优化 反应条件 |
| 文章编号： | 1009-1041（2007）03-0378-04 |
| 收稿时间： | 2006-08-15 |
| 修稿时间： | 2006-11-28 |
Screening and Detecting of Genetically Modified Components in Transgenic Wheat by PCR |
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| LUAN Feng-xi,ZHANG Hong-xiang,BAI Yue.Screening and Detecting of Genetically Modified Components in Transgenic Wheat by PCR[J].Journal of Triticeae Crops,2007,27(3):378-381. |
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| Authors: | LUAN Feng-xi ZHANG Hong-xiang BAI Yue |
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| Institution: | Heilongjiang Entry-Exit Inspection and Quarantine Bureau, Harbi, Heilongjiang 150001 ,China |
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| Abstract: | As the transgenic products being managed strictly in many countries and the great progress achieved in commercial transgenic wheat,it is important to established the detection methods of transgenic wheat.Genomic DNA from wheat was extracted from B73,B72 and B102,wheat lines transferred by HMW gene.Polymerase chain reactions were used to detect the ubiquitin promotor,NOS terminate,and reporter gene of bar and uidA in these transgenic wheat lines.In addition,A wheat endogenous reference gene GAG56D was found at the first time.The primers of GAG56D and ubiquitin genes were designed and PCR conditions were optimized.PCR products were analyzed by agrogel electrophoresis and the expected results were achieved.And the results had been proved by real-time PCR.The lower limit of detection was less than 0.5%.The method can be used for detecting transgenic wheat universally. |
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| Keywords: | Wheat Transgenic PCR Qualitative detection |
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