| 转基因大豆MON89788检测质粒标准分子的构建与应用 |
| |
| 引用本文： | 李飞武,邵改革,邢珍娟,李葱葱,夏蔚,张明.转基因大豆MON89788检测质粒标准分子的构建与应用[J].安徽农业科学,2010,38(23):12330-12333. |
| |
| 作者姓名： | 李飞武 邵改革 邢珍娟 李葱葱 夏蔚 张明 |
| |
| 作者单位： | 吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033;吉林省农业科学院,农业部转基因植物环境安全监督检验测试中心,吉林长春,130033 |
| |
| 基金项目： | 转基因生物新品种培育重大专项 |
| |
| 摘 要： | 目的]构建适用于转基因大豆MON89788检测的质粒标准分子。方法]利用定性PCR和连接、转化等分子克隆技术,将大豆内标准基因lectin、MON89788的3′端特异性序列和5′端特异性序列依次克隆到pMD18-T载体上,获得质粒标准分子pMD-LM3M5,并进行适用性验证。结果]获得了3700bp的质粒标准分子,其中重组DNA片段1029bp。该质粒标准分子的定性PCR检测灵敏度达到10copy。结论]该研究构建的质粒标准分子pMD-LM3M5能替代MON89788基体标准品,用于MON89788大豆及其产品的定性PCR检测。
|
| 关 键 词： | 转基因生物 质粒标准分子 MON89788大豆 转化体特异性检测 |
Construction of the Plasmid Reference Molecule for Detection of Transgenic Soybean MON89788 |
| |
| Institution: | LI Fei-wu et al (Jilin Academy of Agricultural Sciences,National Center for Environmental Safety Inspection of Transgenic Plants,MOA,Changchun,Jilin 130033) |
| |
| Abstract: | Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788. Method]The lectin gene sequence,3'-junction and 5'-junction sequence between host plant DNA and integrated DNA of MON89788 soybean were amplified independently,and the three fragments were cloned into the cloning vector pMD18-T in order through molecular manipulation method to construct pMD-LM3M5,the applicability of the constructed novel PRM was tested. Result] Sequencing confirmation result showed that the PRM was 3 700 bp in length,containing 1 029 bp of recombined DNA fragment. The limits of qualitative detection of the PRM was 10 copy. Conclusion] The PRM constructed in this study was suitable for the identification of MON89788 event. |
| |
| Keywords: | Genetically modified organisms Plasmid reference molecule MON89788 soybean Event-specific detection |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
