摘要
DNA环介导等温扩增技术是一种特异、灵敏、快速的新型基因检测技术。针对玉米表达载体的花椰菜花叶病毒35 S启动子(CaMV35S)的6个区域设计4种特异引物,对LAMP反应的MgSO4、dNTPs、Betaine、内引物、外引物各个成分进行了优化,此外还对LAMP和PCR两种不同方法的特异性进行了比较。建立转基因玉米花椰菜花叶病毒35 S启动子环介导等温扩增技术检测方法,LAMP与PCR相比具有更高的灵敏度。环介导等温扩增技术检测方法具有时间少,成本低,特异性高,检测方法多样等优势,为检测转基因玉米花椰菜花叶病毒35 S启动子提供了一种更加简便快速的方法。
The objective is to develop a loop-mediated isothermal amplification method for detection of CaMV-35S promoter in Genetically Modified maize.Loop-mediated isothermal amplification（LAMP）,a novel nucleic acid amplification method,was developed for the rapid detection of Genetically Modified maize.Cauliflower mosaic virus 35S（CaMV35S） promoter gene was amplified by a set of four primers that recognize six distinct sequences of the target.The optimized conditions for LAMP were studied using different Mg2＋,dNTPs,Betaine and primers concentrations.Furthermore,the sensitivity of LAMP was tested comparing with PCR.The LAMP method can detect CaMV35S promoter from genetically modified maize and their test results were consistent with the results of conventional PCR method.LAMP assay results were found to be more sensitive than the conventional PCR.The LAMP assay is an extremely rapid,highly sensitive,specific method,and will be an effective tool for rapid detection of Genetically Modified maize.
出处
《华北农学报》
CSCD
北大核心
2011年第4期8-14,共7页
Acta Agriculturae Boreali-Sinica
基金
国家转基因专项资助项目(2009ZX08003-009B)
北京市科委项目(KJCX201102003)
