| 程 阳，胡 珅，栾凤侠.小麦内源基因 GAG56D的快速检测技术体系[J].麦类作物学报,2013,33(6):1100 |
| 小麦内源基因 GAG56D的快速检测技术体系 |
| Rapid Detection of Endogenous Gene GAG56D in Wheat by LAMP |
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| DOI：10.7606/j.issn.1009-1041.2013.06.006 |
| 中文关键词: 小麦内源基因 GAG56D 环状等温扩增技术(LAMP) 快速检测 |
| 英文关键词:Endogenous gene of wheat GAG56D Loop-mediated isothermal amplification(LAMP) Rapid detection |
| 基金项目:国家质检总局科研项目(2011B286k)；黑龙江出入境检验检疫局科研项目(2013HK001)。 |
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| 中文摘要: |
| 为了满足转基因小麦检测中对内源参照基因的检测要求，以小麦属特异的内源基因γ-醇溶蛋白基因（ GAG56D）作为目的基因，采用环状等温扩增技术(LAMP)建立了小麦内源基因 GAG56D的快速检测技术体系。针对小麦内源基因 GAG56D特异靶序列的6个区域设计4 条特异性引物，通过显色法进行LAMP检测，并对时间、温度等反应条件及反应灵敏度、特异性、稳定性进行探索。结果表明，该检测体系最适反应温度为63℃，反应时间 60 min，定性检测低限达到0.5%（w/w）。该检测方法具有高度的特异性、灵敏度和稳定性，操作简单、快速。 |
| 英文摘要: |
| The objective of this paper is to establish a rapid detection method for the reference gene in wheat,which is a necessary step in the detection of genetically modified wheat, A loop-mediated isothermal amplification (LAMP) method was developed in this article,targeting to γ- GAG56D, the specific endogenuous gene in wheat. Based on the LAMP method, four specific primers for wheat were designed according to the sequences in six target region of GAG56D, and the reactive conditions were optimized. At last, visual detection could confirm the results of LAMP amplification. The results showed that the optimum temperature and reaction duration was 63℃ and 60 min, respectively. The sensitivity of LAMP detection was 0.5% （w/w）. Results indicated that LAMP was a sensitive, specific, stable and quick method for the detection of GAG56D in wheat. |
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