应用依赖解旋酶DNA扩增技术检测转基因大豆  

Detection of Genetically Modified Soybean by Helicase-Dependent Isothermal DNA Amplification Assay

作　　者：章晶晶[1] 王赞[1] 张翠侠[1] 马超峰[1] 周巍[1,2] 张岩[1] 

机构地区：[1]河北省食品检验研究院,河北省食品安全重点实验室,河北石家庄050071 [2]河北农业大学食品科技学院,河北保定071000

出　　处：《食品科学》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)

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.

关 键 词：依赖解旋酶DNA扩增 转基因大豆 检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]