大豆加工食品中转基因成分多重PCR检测体系的建立

Multiplex PCR for Detection of Transgenic Components in Soybean and Its Derived Food Products

以大豆内源基因 (Lectin)、筛选基因 3 5S启动子 (Cauliflowermosaicvirus 3 5S ,CaMV3 5S)、Nos终止子 (Nopalinesynthase,Nos)和外源基因 (5 enolpyruvylshikimate 3 phosphatesynthase ,Ep sps)为检测对象 ,通过对PCR扩增体系中各引物终浓度及PCR扩增过程中退火温度的探讨 ,研究了不同引物终浓度配比及退火温度对转基因大豆多重PCR检测的影响 ,建立了大豆加工食品中转基因成分多重PCR检测体系。结果表明 ,当各组引物的终浓度分别为 1 0、2 0、2 0、3 0 μmol/L即引物终浓度配比为 1∶2∶2∶3 ,退火温度为 5 5 4℃时 ,所建立的多重PCR检测方法能够有效地检测出大豆中的转基因成分 ,具有特异性好 ,简便 ,快速 ,准确等优点。

To detect transgenic components in genetically modified soybean and its products, endogenous gene Lectin, CaMV35S promoter, Nos terminator and exogenous gene Epsps were chosen for establishing a multiplex polymerase chain reaction method. The PCR conditions for the multiplex PCR analysis were optimized according to the primer concentration and annealing temperature. The results show that the high efficiency PCR system was obtained when the final concentration of the primer is 10, 20, 20, 30μmol/L for Lectin, 35S, Nos and Epspsp respectively, and the annealing temperature is 55.4℃. This method is a sensitive, specific, simple and accurate detection for transgenic component, and thus provided a useful tool for analysis of food products.