豆制品中转基因成分的二重PCR检测体系的优化研究

Optimizing of double PCR system for detecting transgenic ingredients in soybean products

中文关键词:  转基因检测  二重PCR  聚丙烯酰胺凝胶电泳

英文关键词:transgenic detection  double PCR  polyacrylamide gel electrophoresis

中文摘要:

      目的 由于转基因大豆及其制品日益增多, 其安全性亦受到了越来越多消费者的关注。为满足消费者的知情选择权, 本研究建立一种快速检测外源基因的方法。方法 以转基因豆制品中存在的CaMV35S启动子和NOS终止子为检测目标, 优化了二重PCR反应体系和条件, 包括引物比例和酶的用量; 运用聚丙烯酰胺凝胶电泳和琼脂糖凝胶电泳对PCR产物进行检测, 并对聚丙烯凝胶电泳的银染条件进行探索研究。结果 建立了运用聚丙烯酰胺凝胶电泳更好地区分出外源基因CaMV35S启动子和NOS终止子的目的条带的方法。结论 本实验中的二重PCR和聚丙烯酰胺凝胶电泳方法适用于对转基因豆制品中这两个外源基因的检测, 为转基因豆制品检测提供了指导。

英文摘要:

      Objective Account of the increasing amount of transgenic soybean and soybean products, the safety problem has received more and more attention from consumers. In order to satisfy consumers’ right of informed choice, a rapid method of detecting genetically modified products was established in this study. Methods CaMV35S promoter and NOS terminator in soybean products were taken as detection target. The double PCR reaction system and conditions were optimized, including primer ratio and enzyme amount. Agarose gel and polyacrylamide gel electrophoresis were applied to analyze PCR products, and the silver staining conditions for polyacrylamide gel electrophoresis were explored. Results The polyacrylamide gel electrophoresis was established and could distinguish CaMV35S promoter and NOS terminator better than agarose gel. Conclusion The results showed that double PCR and polyacrylamide gel electrophoresis in this experiment were suitable for the analytic detection of two foreign genes in transgenic soybean products, and it could be a guidance for detecting transgenic food.