大豆、油菜籽及大豆油中外源基因检测的灵敏度、准确度研究

Sensitivity and Accuracy Detections for Exotic Gene in Soybean, Rape Seed and Soybean Oil

本研究针对转基因产品中转入的目的基因以及启动子、终止子,设计了相应引物35SCP142、CPNOS165。通过对引物的调整,使PCR扩增产物的大小控制在100～170bp,从而提高了检测灵敏度。在25μl的PCR反应体系中,可以检测到0.01ng的外源DNA。为了检测以大豆为原料的油脂类产品中的外源基因,我们又设计了NEST-PCR的两对引物35SCP318和35SCP114,并检测到了外源基因的片断。对新设计的引物进行准确度分析,即在不同的样品(包括转基因或非转基因产品)中进行检测,并经测序验证,新设计的引物35SCP142,CPNOS165及NEST-PCR的引物都有高特异性,可用于含有相应外源基因的产品检测。

Referring to the sequences of some promoter, terminator and exotic gene, the primers 35SCP142 and CPNOS 165 were designed. Sensitivity has been obviously improved by optimizing the primers which could make the length of amplicon shorten from 170bp to 100bp. Up to 0.01ng exotic gene could be detected in a 25 μl volume PCR amplification system. In order to detect the exotic gene in soybean oil, two primers 35SCP318 and 35SCP114 for NEST-PCR （nest polymerase chain reaction） have been designed, with which DNA fragments of exotic gene have been amplified and detected. Accuracy of the new primers also have been estimated by being applied to several kinds of products, including GMOs and NON-GMOs. Verfied by sequencing, new primers namely 35SCP142, CPNOS 165 and NEST-PCR primers posses high specificities, and could be used for exotic genes detection.