大豆及腐竹中转基因成分的多重PCR分析

Multiplex polymerase chain reaction(MPCR) analysis of genetically modified ingredients of the soybean and beanthread

采用CTAB法提取大豆干样、新鲜毛豆及腐竹中的总DNA,内源LECTIN基因扩增结果均为阳性,表明提取到的DNA中不存在抑制PCR的物质.应用花椰菜花叶病毒(Cauliflowermosaicvirus,CaMV)35S启动子和根癌农杆菌胭脂碱合成酶基因(NOS)终止子的二重PCR分析,筛选到7个含转基因成分的样品.半嵌套式PCR检测结果表明阳性样品中的外源目的基因为来自根癌农杆菌CP4菌株的EPSPS基因.最后成功进行了扩增内源LECTIN基因、CaMV35S启动子和NOS终止子的三重PCR分析,以及内源LECTIN基因和EPSPS基因的二重PCR分析,得到预期结果.多重PCR方法具有快速、简便、准确等特点,在植物产品的转基因成分检测上具有重要的应用价值.

The DNAs of dry soybean, fresh soybean and beanthread were extracted by CTAB method. The detection results of inner LECTIN gene were positive, which showed that there were no substance restraining polymerase chain reaction(PCR) in the DNAs. Seven samples containing genetically modified ingredients were found by duplex PCR of Cauliflower mosaic virus(CaMV) 35S promoter and Agrobacterium tumefaciens nopaline synthase(NOS) terminator. The heminested PCR results showed that the foreign genes of the seven positive samples were EPSPS genes from A.tumefaciens strain CP4. The triplex PCR of inner LECTIN gene, CaMV 35S promoter and NOS terminator, duplex PCR of inner LECTIN and EPSPS gene were carried on successfully, and obtained the same results as expected. The multiplex PCR was quick, simple and practical, and might play an important role in the detection of genetically modified ingredient in plant products.