摘要
目的:基于依赖解旋酶DNA恒温扩增(helicase-dependent isothermal DNA amplifi cation,HDA)技术,建立快速检测转基因大豆的方法。方法:采用以CaMV35S、NOS、CP4-EPSPS 3种外源基因和内源基因Lectin(大豆凝集素基因)为目的片段设计4对特异引物,建立反应体系,通过HDA方法对内源基因和3种外源基因的检测特异性和灵敏度进行实验,并对结果进行同源性分析。结果:建立了转基因大豆HDA检测法,检测特异性良好,3种外源基因的检出限为0.2%。结论:转基因大豆的HDA检测方法具有普通聚合酶链式反应的特异、灵敏等特点,并且对仪器要求更低,极适合基层实验室使用,具有广阔的应用前景。
Objective： To establish a new rapid method to detect genetically modifi ed soybean based on helicase-dependent isothermal DNA amplification（HDA）. Methods： With four highly specific sets of primers synthesized to target genes including Lectin in soybean and Ca MV35S,NOS and CP4-EPSPS foreign genes,PCR reaction system was established. The specifi city and sensitivity of the PCR method were tested based on HDA and homology analysis was performed. Results： The helicase-dependent isothermal DNA amplifi cation method for the rapid detection of genetically modifi ed soybean has been established. The detection limits of Ca MV35S,NOS and CP4-EPSPS genes in genetically modifi ed soybeans by HDA were all 0.2%. Conclusion： HDA is a rapid,user-friendly,specifi c and sensitive method for the detection of genetically modifi ed soybean,and is very suitable for use in grassroots laboratories with a broad prospect.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2016年第4期164-168,共5页
Food Science
基金
国家自然科学基金面上项目(31401584)
关键词
依赖解旋酶DNA扩增
转基因大豆
检测
helicase-dependent isothermal DNA amplifi cation（HDA）
genetically modifi ed soybean
detection
